A Modified 1H-NMR Quantification Method of Ephedrine Alkaloids in Ephedrae Herba Samples.

A previous 1H-NMR method allowed the quantification of ephedrine alkaloids; however, there were some disadvantages. The cyclized derivatives resulted from the impurities of diethyl ether were identified and benzene was selected as the better extraction solvent. The locations of ephedrine alkaloids were confirmed with 2D NMR. Therefore, a specific 1H-NMR method has been modified for the quantification of ephedrine alkaloids. Accordingly, twenty Ephedrae Herba samples could be classified into three classes: (I) E. sinica-like species; (II) E. intermedia-like species; (III) others (lower alkaloid contents). The results indicated that ephedrine and pseudoephedrine are the major alkaloids in Ephedra plants, but the concentrations vary greatly determined by the plant species and the collection locations.

A water-soluble nucleolin aptamer-paclitaxel conjugate for tumor-specific targeting in ovarian cancer.

Paclitaxel (PTX) is among the most commonly used first-line drugs for cancer chemotherapy. However, its poor water solubility and indiscriminate distribution in normal tissues remain clinical challenges. Here we design and synthesize a highly water-soluble nucleolin aptamer-paclitaxel conjugate (NucA-PTX) that selectively delivers PTX to the tumor site. By connecting a tumor-targeting nucleolin aptamer (NucA) to the active hydroxyl group at 2' position of PTX via a cathepsin B sensitive dipeptide bond, NucA-PTX remains stable and inactive in the circulation. NucA facilitates the uptake of the conjugated PTX specifically in tumor cells. Once inside cells, the dipeptide bond linker of NucA-PTX is cleaved by cathepsin B and then the conjugated PTX is released for action. The NucA modification assists the selective accumulation of the conjugated PTX in ovarian tumor tissue rather than normal tissues, and subsequently resulting in notably improved antitumor activity and reduced toxicity.

Present Advances and Future Perspectives of Molecular Targeted Therapy for Osteosarcoma.

Osteosarcoma (OS) is a bone cancer mostly occurring in pediatric population. Current treatment regime of surgery and intensive chemotherapy could cure about 60%-75% patients with primary osteosarcoma, however only 15% to 30% can be cured when pulmonary metastasis or relapse has taken place. Hence, novel precise OS-targeting therapies are being developed with the hope of addressing this issue. This review summarizes the current development of molecular mechanisms and targets for osteosarcoma. Therapies that target these mechanisms with updated information on clinical trials are also reviewed. Meanwhile, we further discuss novel therapeutic targets and OS-targeting drug delivery systems. In conclusion, a full insight in OS pathogenesis and OS-targeting strategies would help us explore novel targeted therapies for metastatic osteosarcoma.

Mass-transport-controlled, large-area, uniform deposition of carbon nanofibers and their application in gas diffusion layers of fuel cells.

The effect of mass transport on the growth characteristics of large-area vapor-grown carbon nanofibers (CNFs) was investigated by adjusting the substrate deposition angle (α). The catalyst precursor solution was coated onto one side of a 2D porous carbon paper substrate via a decal printing method. The results showed that the CNFs were grown on only one side of the substrate and α was found to significantly affect the growth uniformity. At α = 0°, the growth thickness, the density, the microstructure and the yield of the CNF film were uniform across the substrate surface, whereas the growth uniformity decreased with increasing α, suggesting that the large-area CNF deposition processes were mass-transport-controlled. Computational fluid dynamics simulations of the gas diffusion processes revealed the homogeneous distributions of the carbon-source-gas concentration, pressure, and velocity near the substrate surface at α = 0°, which were the important factors in achieving the mass-transport-limited uniform CNF growth. The homogeneity of the field distributions decreased with increasing α, in accordance with the variation in the growth uniformity with α. When used as a micro-porous layer, the uniform CNF film enabled higher proton exchange membrane fuel cell performance in comparison with commercial carbon black by virtue of its improved electronic and mass-transport properties confirmed by the electrochemical impedance spectroscopy results.

[Effect of supplementing qi, blood-activating and kidney-nourishing therapy on postoperative recovery in patients with lumber disc herniation].

OBJECTIVE: To investigate the effect of supplementing qi, activating blood circulation and tonifying kidney therapy on the postoperative outcomes of patients undergoing lumber intervertebral disc herniation. METHODS: From January 2010 to May 2012, 120 patients with lumbar intervertebral disc herniation undergoing surgical treatment in Nanfang hospital were randomized into two equal groups to receive routine therapy (control group) and additional treatment with Yiqi Houxue Bushen Decoction (treatment group). The effect of the interventions was evaluated by assessing the Visual Analogue Scale(VAS), Japanese Orthopedic Association Scores (JOA), WHO Quality of Life-BREF (WHOQOL-BREF), length of hospital stay and adverse event. RESULTS: All the 120 patients were followed up and analyzed. Significant differences were found between the treatment and control groups in VAS, JOA Scores, and WHOQOL-BREF (P<0.01) at 2, 4, and 8 week and at 6 and 12 months after the surgery. At 6 and 12 months postoperatively, the JOA Scores (P<0.01), but not the VAS and WHOQOL-BREF, differed significantly between the two groups. CONCLUSION: Blood-activating and kidney-nourishing therapy is effective in promoting postoperative recovery and helps reduce the clinical symptoms and minimize the adverse events in patients undergoing surgery for lumber intervertebral disc herniation.

[Value of X-ray combined with ultrasound in the diagnosis and treatment of solitary osteochondroma].

OBJECTIVE: To explore the most appropriate imaging examinations for solitary osteochondroma in primary hospitals. METHODS: A retrospective evaluation was performed to analyze the imaging examinations (including X-ray, CT, and ultrasound) of 62 patients undergoing surgeries for solitary osteochondroma in terms of the diagnostic results and preoperative expenditure of the patients. RESULTS: X-ray and CT have a high diagnostic value for solitary osteochondroma, but X-ray cannot display cartilage and lesions in surrounding soft tissues. Ultrasound is limited in diagnosis of solitary osteochondroma, but performed well as CT in displaying cartilage and lesions in surrounding soft tissues. There was no statistical difference between combined X-ray and ultrasound and CT in diagnosis of solitary osteochondroma (P>0.05), but the former had a lower cost (P<0.05). CONCLUSION: X-ray combined with ultrasound has almost comparable value with CT in the diagnosis of solitary osteochondroma and is more practical for application in primary hospitals.

Synthesis of 2-(1,5-diaryl-1,4-pentadien-3-ylidene)-hydrazinecarboximidamide hydrochloride catalyzed by p-dodecylbenzenesulfonic acid in aqueous media under ultrasound irradiation.

Amidinohydrazone compounds are very important synthetic intermediates and can serve as versatile precursors in synthesis of many natural products and drug molecules. The use of ultrasound, p-dodecylbenzenesulfonic acid (DBSA) and water as solvent improved the synthesis of different 2-(1,5-diaryl-1,4-pentadien-3-ylidene)-hydrazinecarboximidamide hydrochlorides. The best reaction conditions for the condensation of 1,5-diphenyl-1,4-pentadien-3-one with aminoguanidine hydrochloride were as follows: 1,5-diphenyl-1,4-pentadiene-3-one (1, 1 mmol), aminoguanidine hydrochloride (1.1 mmol), DBSA (0.5 mmol), water 10 mL, reaction temperature 25-27°C, irradiation frequency 25 kHz. 2a was achieved in 94% yield within 2h. The other seven amidinohydrazones were obtained in 84-94% yield within 2-3h under the same conditions. Compared to the method involving catalysis by hydrochloric acid in refluxing EtOH, the advantages of present procedure are milder conditions, shorter reaction times, higher yields, and environmental friendly conditions, which make it a useful strategy for the synthesis of analogues.

Arthroscopically assisted reduction with volar plating or external fixation for displaced intra-articular fractures of the distal radius in the elderly patients.

Twenty-eight patients older than 70 years with AO type C fracture of the distal radius were treated with arthroscopically assisted reduction combined with volar plating or external fixation. The patients were followed up for an average of 24.9 +/- 16.1 months. The average score was 80.1 +/- 10.5 according to the modified system of Green and O'Brien. Eight patients had an excellent result, 11 had a good result, seven had a fair result, and two had a poor result. Twenty-three patients were able to return to their previous activities level or occupation without any restriction. On the basis of these results, we concluded that arthroscopically assisted reduction combined with volar plating or external fixation is one of the useful options for the treatment of a displaced intra-articular fracture of the distal radius in elderly patients who are physiologically young or active.

Absence of cutaneous TNFalpha-producing CD4+ T cells and TNFalpha may allow for fibrosis rather than epithelial cytotoxicity in murine sclerodermatous graft-versus-host disease, a model for human scleroderma.

Graft-versus-host disease (GVHD) is a complication of hematopoietic cell transplantation and is a major source of morbidity and mortality. Two main forms of GVHD occur: cytotoxic GVHD (Cyt GVHD), in which TNFalpha is a critical cytokine in epithelial injury, and sclerodermatous GVHD (Scl GVHD), in which TGFbeta plays a major role in fibrosis. To understand the critical early events in GVHD and scleroderma, we are studying a murine model that uses differences in minor histocompatibility antigens to generate Scl GVHD. We asked the question: what is the immune environment in this model that promotes fibrosis rather than the epithelial injury of Cyt GVHD? We found that in Scl GVHD, cutaneous CD4+ T cells produced IFNgamma and IL-2 but not TNFalpha, also absent by gene array analysis. The role of cutaneous CD4+ T cells in Scl GVHD may not be an active process through production of TGFbeta, but may rather be a passive one due to lack of antigen-presenting cell (APC) support for CD4+ T cells and failure to produce TNFalpha, a potent inhibitor of TGFbeta-induced fibrosis as well as inducer of keratinocyte apoptosis. These APC-T cell interactions and the cytokine environment promote fibrosis rather than cytotoxic epithelial injury in skin in Scl GVHD.

[Pathological characteristics of dilated hearts with sole myocardial wall damage in recipients].

OBJECTIVE: It is difficult to differentiate the causes of dilated cardiomyopathy only by clinical evaluation and image analysis. Pathomorphologic examinations on diseased hearts may help to improve the diagnosis accuracy. METHODS: Fifty-six extransplanted hearts from June, 2004 to June, 2006 were examined. Gross and histopathological findings were recorded, photographed and final pathological diagnosis was compared to clinical diagnosis. RESULTS: Dilations were caused by sole myocardial wall damage in 38 (67.9%) of the 56 patients, including 19 primary dilated cardiomyopathy, 9 arrhythmogenic right ventricular cardiomyopathy, 1 non-compaction cardiomyopathy, 6 ischemic cardiomyopathy, 1 alcoholic cardiomyopathy, 1 hypertensive cardiomyopathy and 1 giant cell myocarditis. The clinical and pathological diagnoses were different in 15 cases (39.5%). The most discrepancies were arrhythmogenic right ventricular cardiomyopathy (77.8%), ischemic cardiomyopathy (83.3%), and giant cell myocarditis (100%). CONCLUSIONS: This pathological study of recipient hearts showed a high portion of patients with arrhythmogenic right ventricular cardiomyopathy and ischemic cardiomyopathy were misdiagnosed as primary cardiomyopathy. Correct diagnosis of primary cardiomyopathy needs to rule out possible secondary causes of myocardial dilation.

Fibroblast expression of the coactivator p300 governs the intensity of profibrotic response to transforming growth factor beta.

OBJECTIVE: Transforming growth factor beta (TGFbeta) induces profibrotic responses in normal fibroblasts, and plays a fundamental role in the pathogenesis of fibrosis in scleroderma (systemic sclerosis [SSc]). The intensity of cellular responses elicited by cytokines is modulated by transcriptional coactivators such as the histone acetylase p300. The objective of these studies was to delineate the physiologic role of p300 in Smad-dependent profibrotic responses elicited by TGFbeta. METHODS: Ectopic p300 was transiently expressed in normal dermal fibroblasts. Cellular p300 levels were suppressed using p300-specific ribozymes. The regulation of gene expression was examined by transient transfection assays, Northern blotting, and immunoblot analysis. The expression of p300 in normal and scleroderma fibroblasts was evaluated by confocal microscopy and immunoblotting, and p300 levels in skin from mice with experimental scleroderma were assessed by immunohistochemistry. RESULTS: In normal fibroblasts, TGFbeta induced an increase in the levels of p300. Forced expression of ectopic p300 in these cells dramatically enhanced the magnitude of TGFbeta responses, whereas selective depletion of p300 using ribozyme resulted in abrogation of TGFbeta-induced collagen synthesis and promoter activity. Furthermore, TGFbeta lost its ability to induce Smad-dependent transcription in p300-depleted fibroblasts. These responses could be fully rescued with ectopic p300. Abrogation of Smad-mediated TGFbeta signaling was not due to alterations in the levels or the ligand-dependent phosphorylation or intracellular trafficking of endogenous Smads. Immunohistochemical analysis demonstrated substantially increased p300 expression in lesional skin from mice with chronic graft-versus-host disease, an animal model of scleroderma. Furthermore, levels of p300 were 2-3-fold higher in cultured fibroblasts derived from SSc patients than in fibroblasts from matched normal controls. CONCLUSION: These results establish, for the first time, that the coactivator histone acetylase p300, itself a target of TGFbeta regulation, is an essential component of the cellular TGFbeta signal transduction pathways mediating stimulation of collagen synthesis in fibroblasts. Since the cellular abundance of p300 appears to govern the intensity of profibrotic responses elicited by TGFbeta, elevated p300 expression in lesional tissue may contribute to the progression of skin fibrosis in scleroderma.

Successful auricle replantation via microvascular anastomosis 10 h after complete avulsion.

OBJECTIVE: To report a case of successful total auricle replantation 10 h after complete amputation. MATERIAL AND METHODS: Replantation was achieved by means of microvascular anastomosis and other therapies, and assisted by careful nursing. The patient's auricle was completely cut off with a sharp blade by an assailant, who retained it for 5 h. Having been retrieved, it was then preserved in ice for 5 h. Using microscopy, an artery and vein were found at the confluences of the upper and middle and lower and middle parts of the amputated auricle and head wound, respectively An end-to-end anastomosis was performed on these vessels. After operation, the following treatments were used: drainage; reopening of the drainage channels with a needle: flushing or soaking with heparin sodium solution; controlling infection and coagulation; increasing blood volume: dilating vessels; and special nursing. RESULTS: The replanted auricle survived with a normal contour and a very favorable esthetic appearance. CONCLUSIONS: The neat, uncontaminated wound margins and the fact that the amputated auricle was preserved in ice, even though it had been amputated 10 h before the operation, were prerequisites for successful auricle replantation. The microvascular anastomosis technique played a very important role in the survival of the amputated auricle. The postoperative treatment, observation and nursing were difficult but vital aspects of the procedure. The establishment of an effective venous return was a sign of the survival of the amputated auricle.

Inhibition of monocytic interleukin-12 production by Candida albicans via selective activation of ERK mitogen-activated protein kinase.

Our previous data demonstrated that live Candida albicans inhibits interleukin-12 (IL-12) production by human monocytes. Here we explored whether C. albicans inhibits IL-12 via a released factor and whether the inhibition is mediated via mitogen-activated protein kinase (MAPK) regulation. Supernatant fluids were obtained from cultured C. albicans (SC5314) as well as cultured Saccharomyces cerevisiae after 20 h of incubation. At 2 h postincubation of monocytes with heat-killed C. albicans (HKCA) (2:1) to stimulate IL-12, concentrated fungal supernatant fluids were added and incubated for an additional 20 h. The present data show that, unlike supernatant fluids obtained from S. cerevisiae, the C. albicans supernatant fluids significantly suppressed IL-12 production induced by HKCA. This suggested that the inhibition is Candida specific. A preliminary biochemical analysis revealed that this secretory IL-12 inhibitory factor is glycoprotein in nature. The inhibitory activity had no effect on the phagocytosis of yeasts. Supernatant fluids from C. albicans markedly induced the phosphorylation of ERK44/42 MAPK, but not p38 and SAPK, 1 min after they were added to monocytes. To test if the induction of ERK44/42 MAPK was central to the IL-12 inhibition, we used gamma interferon (IFN-gamma) (1 ng/ml) plus lipopolysaccharide (LPS) (100 ng/ml) to stimulate IL-12 production by monocytes. The inhibition of ERK MAPK by the specific inhibitor PD 98059 significantly reduced phospho-ERK44/42 MAPK levels induced by C. albicans supernatant fluids in the IFN-gamma-plus-LPS-driven monocytes. Concomitantly, PD 98059 reversed the IL-12 inhibitory activity of the C. albicans supernatant (P < 0.01). These data indicate that C. albicans can inhibit IL-12 production by secreting an ERK44/42 MAPK-stimulating factor and thus can attenuate effective immune responses.

[Surgical repair of major vascular injuries in the extremities complicated by other injuries: retrospective analysis of 67 cases].

OBJECTIVE: To retrospectively analyze the surgical repair of injured major vessels in the extremities complicated by multiple other injuries. METHODS: According to different time lengths of sustaining ischemia, sites of the major vascular injuries and conditions of other injuries, 91 operations for end-to-end anastomosis, vessel grafting and repair, ligation and shunting respectively were performed on the 67 patients. RESULTS: Except for 2 fatal cases (one due to preoperative brain hernia and hemorrhagic shock, the other to thyroid crisis during operation), all other cases survived the operations. Vascular crisis occurred in 3 cases (2 arterial spasm and 1 venous crisis) 6, 7 and 12 h respectively after the operations, which were relieved by appropriate treatments and surgical explorations. One patient developed pneumonia and acute renal failure with polyuria at postoperative days 7 and 4 respectively, but full recovery was achieved. CONCLUSIONS: For patients with major vascular injuries in the extremity complicated by multiple other injuries, timely revascularization of the extremities should be performed during or immediately after emergency operations of the vital organs. Particular attention should be given to the diagnosis and repair of closed major vascular injuries in such patients, who need to be monitored for blood coagulation during operation and should receive due post-operative anti-coagulation treatment.

[Peripheral nerve regeneration under immunosuppression].

OBJECTIVE: To discuss peripheral nerve regeneration under immunosuppression. METHODS: Current research trends about relationship between peripheral nerve injury and immunoreaction, the experimental result of nerve regeneration after using various immunosuppressors, and the clinical findings after human allogenous hand transplantation were extensively reviewed. RESULTS: Peripheral nerve regeneration was accelerated under immunosuppression. CONCLUSION: Peripheral nerve injury may induce immunoreaction, which inhibit nerve regeneration and function recovery.