Enzymology, Histological and Ultrastructural Effects of Ar-Turmerone on Culex pipiens pallens Larvae.

Our previous article demonstrated that ar-turmerone ((6S)-2-methyl-6-(4-methylphenyl)-2-hepten-4-one) extracted from Curcuma longa L. has a significant larvicidal activity against the fourth instar larvae of Culex pipiens pallens. To reveal the effects of ar-turmerone on C. pipiens pallens larvae, light microscopy and transmission electron microscopy were used to observe the histological and ultrastructure changes in muscle and digestive tissues of fourth instar larvae. It was also revealed by detecting the activity of the acetylcholinesterase (AChE) enzyme and three detoxifying enzymes, including carboxylesterase (CarE), glutathione-S-transferase (GST) and Cytochrome P450 monooxidases (P450). The observation under the light microscope showed that the larvae displayed a disruption of myofibril in ventral muscle cells, the disappearance of nucleolus in the malpighian tubule cells, and the exfoliation of the brush border in midgut epithelial cells, 24 h after treatment. The observation under the transmission electron microscope displayed disorganized Z-lines in the ventral muscle cells, and dissolved membrane of mitochondria, nuclear and endoplasmic reticulum in abdominal cells. The enzymatic activity results showed that ar-turmerone significantly increased the level of detoxifying enzymes, while the activity of AChE was not obviously affected. All the results suggest that the larvicidal mechanism of ar-turmerone is estimated to be stomach poison and the active sites might be the muscle and digestive tissues, and the mode of action of ar-turmerone may be unrelated to AChE.

Enrichment of soil rare bacteria in root by an invasive plant Ageratina adenophora.

The assembly of the root-associated microbiome provides mutual benefits for the host plant and bacteria in soils. It is interesting how invasive plants interact with the local soil microbial community and establish the soil bacterial community in the endosphere of these plants in the short term. In this study, we compared the bacterial community in the rhizosphere with that in the root endosphere of an invasive plant, Ageratina adenophora, using high-throughput sequencing. The results showed that the roots of A. adenophora selectively accumulated the genera Clostridium and Enterobacter, which are rarely distributed in the rhizosphere. This selective accumulation caused a switch in the bacterial composition at the phylum level from Bacteroidetes predominant in the rhizosphere to Proteobacteria dominant in the root endosphere of A. adenophora. Our data indicated the potential existence of a highly conserved signal recognition in which hosts, either invasive or native, enrich the endosphere bacteria, such as Clostridium, Enterobacter, etc., from the rhizosphere. Moreover, the accumulated bacteria were physiologically and genetically different at the strain level and displayed distinct roles in growth between invasive and native plants. The assembly of the bacterial community in the roots may be an advantageous strategy for A. adenophora in competition with native plants.

The potential impact of invasive woody oil plants on protected areas in China under future climate conditions.

Biodiesel produced from woody oil plants is considered a green substitute for fossil fuels. However, a potential negative impact of growing woody oil plants on a large scale is the introduction of highly invasive species into susceptible regions. In this study, we examined the potential invasion risk of woody oil plants in China's protected areas under future climate conditions. We simulated the current and future potential distributions of three invasive woody oil plants, Jatropha curcas, Ricinus communis, and Aleurites moluccana, under two climate change scenarios (RCP2.6 and RCP8.5) up to 2050 using species distribution models. Protected areas in China that will become susceptible to these species were then identified using a spatial overlay analysis. Our results showed that by 2050, 26 and 41 protected areas would be threatened by these invasive woody oil plants under scenarios RCP2.6 and RCP8.5, respectively. A total of 10 unique forest ecosystems and 17 rare plant species could be potentially affected. We recommend that the invasive potential of woody oil plants be fully accounted for when developing forest-based biodiesel, especially around protected areas.

Altered regional and circuit resting-state activity in patients with occult spastic diplegic cerebral palsy.

BACKGROUND: Very few studies have been made to investigate functional activity changes in occult spastic diplegic cerebral palsy (SDCP). The purpose of this study was to analyze whole-brain resting state regional brain activity and functional connectivity (FC) changes in patients with SDCP. METHODS: We examined 12 occult SDCP and 14 healthy control subjects using resting-state functional magnetic resonance imaging. The data were analyzed using Resting-State fMRI Data Analysis Toolkit (REST) software. The regional homogeneity (ReHo), amplitude of low-frequency fluctuations (ALFF), and whole brain FC of the motor cortex and thalamus were analyzed and compared between the occult SDCP and control groups. RESULTS: Compared with the control group, the occult SDCP group showed decreased ReHo regions, including the bilateral frontal, parietal, and temporal lobes, the cerebellum, right cingulate gyrus, and right lenticular nucleus, whereas an increased ReHo value was observed in the left precuneus, calcarine, fusiform gyrus, and right precuneus. Compared with the control group, no significant differences in ALFF were noted in the occult SDCP group. With the motor cortex as the region of interest, the occult SDCP group showed decreased connectivity regions in the bilateral fusiform and lingual gyrus, but increased connectivity regions in the contralateral precentral and postcentral gyrus, supplementary motor area, and the ipsilateral postcentral gyrus. With the thalamus being regarded as the region of interest, the occult SDCP group showed decreased connectivity regions in the bilateral basal ganglia, cingulate, and prefrontal cortex, but increased connectivity regions in the bilateral precentral gyrus, the contralateral cerebellum, and inferior temporal gyrus. CONCLUSIONS: Resting-state regional brain activities and FC changes in the patients with occult SDCP exhibited a special distribution pattern, which is consistent with the pathology of the disease.

Comparative analysis of curative effect of bone marrow mesenchymal stem cell and bone marrow mononuclear cell transplantation for spastic cerebral palsy.

BACKGROUND: Bone marrow mesenchymal stem cells (BMMSCs) and bone marrow mononuclear cells (BMMNCs) are both used to treat spastic cerebral palsy. However, the differences in therapeutic effect remain unknown. METHODS: A total of 105 patients with spastic cerebral palsy were enrolled and randomly assigned to three groups: the BMMSC group, the BMMNC group and the control group. Patients in both transplantation groups received four intrathecal cell injections. Patients in the control group received Bobath therapy. The gross motor function measure (GMFM) and the fine motor function measure (FMFM) were used to evaluate the therapeutic efficacy before transplantation and 3, 6, and 12 months after transplantation. RESULTS: Three months after cell transplantation, scores in the A dimension of GMFM and the A and C dimensions of FMFM scores in the BMMSC group are all higher than those of the BMMNC and the control groups (P < 0.05). Six months after cell transplantation, scores in the A, B dimensions of GMFM and the A, B, C, D, and E dimensions of FMFM scores in the BMMSC group are higher than those of the BMMNC and the control groups (P < 0.05). Twelve months after cell transplantation, scores in the A, B, and C dimensions of GMFM and the A, B, C, D, and E dimensions of FMFM scores in the BMMSC group are all higher than those of the BMMNC and the control groups (P < 0.05). No obvious adverse effects were investigated during follow-up. CONCLUSIONS: BMMSC transplantation for the treatment of cerebral palsy is safe and feasible, and can improve gross motor and fine motor function significantly. In addition, compared with BMMNC, the motor function of children improved significantly in terms of gross motor and fine motor functions.

Anatomical mechanism of spontaneous recovery in regions caudal to thoracic spinal cord injury lesions in rats.

BACKGROUND: The nerve fibre circuits around a lesion play a major role in the spontaneous recovery process after spinal cord hemisection in rats. The aim of the present study was to answer the following question: in the re-control process, do all spinal cord nerves below the lesion site participate, or do the spinal cord nerves of only one vertebral segment have a role in repair? METHODS: First we made a T7 spinal cord hemisection in 50 rats. Eight weeks later, they were divided into three groups based on distinct second operations at T7: ipsilateral hemisection operation, contralateral hemisection, or transection. We then tested recovery of hindlimbs for another eight weeks. The first step was to confirm the lesion had role or not in the spontaneous recovery process. Secondly, we performed T7 spinal cord hemisections in 125 rats. Eight weeks later, we performed a second single hemisection on the ipsilateral side at T8-T12 and then tested hindlimb recovery for another six weeks. RESULTS: In the first part, the Basso, Beattie, Bresnahan (BBB) scores and the electrophysiology tests of both hindlimbs weren't significantly different after the second hemisection of the ipsilateral side. In the second part, the closer the second hemisection was to T12, the more substantial the resulting impairment in BBB score tests and prolonged latency periods. CONCLUSIONS: The nerve regeneration from the lesion area after hemisection has no effect on spontaneous recovery of the spinal cord. Repair is carried out by all vertebrae caudal and ipsilateral to the lesion, with T12 being most important.

Acaricidal activity of compounds from Cinnamomum camphora (L.) Presl against the carmine spider mite, Tetranychus cinnabarinus.

BACKGROUND: Tetranychus cinnabarinus (Boisduval) is one of the most important, highly polyphagous pests of a wide range of field and greenhouse crops throughout the world. The control of this mite is still based primarily on the use of synthetic chemical pesticides. In this study, we screened eight plant extracts from China and evaluated the natural compounds showing acaricidal properties from the plant extract, considering their potential use as an alternative to synthetic pesticides. RESULTS: In bioassay screening assays, the Cinnamomum camphora (L.) Presl extract showed significantly greater acaricidal activity against T. cinnabarinus than the other seven plant extracts tested. Five compounds were identified from the C. camphora extract via repeated column chromatography and gas chromatography-mass spectrometry analysis. All the compounds presented acaricidal activity, with 2,4-di-tert-butylphenol and ethyl oleate exhibiting the greatest activity. At 7 days after treatment in a potted seedling experiment, the LC50 values of 2,4-di-tert-butylphenol and ethyl oleate were found to be 1850.94 and 2481.65 mg kg(-1) respectively. Microscopic observations showed that the mites displayed the symptomology of poisoning. CONCLUSION: These results demonstrated that the C. camphora extract and its two active components show the potential to be developed as new natural acaricides for controlling carmine spider mites.

Effect of umbilical cord mesenchymal stromal cells on motor functions of identical twins with cerebral palsy: pilot study on the correlation of efficacy and hereditary factors.

BACKGROUND AIMS: The objective of this study was to compare the impact of umbilical cord-derived mesenchymal stromal cell (UCMSC) transplantation on the motor functions of identical twins with cerebral palsy (CP) and to analyze the correlation between the efficacy and hereditary factors. METHODS: Eight pairs (16 individuals) of identical twins with CP were recruited and received allogenic UCMSC transplantation by means of subarachnoid injection. The gross motor function measure (GMFM) and the fine motor function measure (FMFM) were performed before and 1 and 6 months after the treatment to analyze the results of individuals before and after the therapy, between two individuals of an identical twin and among twin pairs. Repeated-measured data variance was used to analyze the GMFM and FMFM scores of patients before and 1 and 6 months after the therapy. RESULTS: Eight pairs (16 individuals) of children with CP had significant improvement in the GMFM at the end of the 1st and 6th months after the therapy compared with that before the therapy, whereas the amelioration of the FMFM was not statistically significant. The improvements in motor functions between two individuals of an identical twin but not among twin pairs were correlated. CONCLUSIONS: UCMSC transplantation significantly improves GMFM in children with CP; motor function improvements in the GMFM between two individuals of an identical twin were closely correlated, but improvements among twin pairs were not correlated. We hypothesize that hereditary factors contribute to the mechanisms of UCMSC transplantation in motor function improvement in children with CP.

Clinical observation of umbilical cord mesenchymal stem cell transplantation in treatment for sequelae of thoracolumbar spinal cord injury.

BACKGROUND: Umbilical cord mesenchymal stem cells (UCMSCs) have a considerable advantage and potential in treating for central nervous system diseases and have become a novel alternative treatment for spinal cord injury. This study aims to compare the neurological function outcome of stem cell transplantation, rehabilitation therapy, and self-healing for sequelae of spinal cord injury. METHODS: Thirty-four cases of thoracolumbar spinal cord injury were randomly divided into three groups: the stem cell transplantation group was given CT-guided UCMSC transplantation twice; the rehabilitation group received rehabilitation therapy; and the blank control group did not receive any specific treatment. AIS grading, ASIA scoring, the manual muscle strength and muscle tension scale, and the Barthel index were used to evaluate the clinical outcome. Urodynamic examination was also performed for patients in the UCMSC group and the rehabilitation therapy group. RESULTS: Seven of the ten patients in the UCMSC group had significant and stable improvement in movement, self-care ability, and muscular tension; five of the forteen patients (36%) in the rehabilitation group also had certain improvement in these aspects. Urodynamic examination demonstrated that patients in the UCMSC group exhibited an increase in maximum urinary flow rate and maximum bladder capacity, as well as a decrease in residue urine volume and maximum detrusor pressure. The rehabilitation group exhibited decreased maximum bladder capacity, but no perceptible change in maximum urinary flow rate, residue urine volume or maximum detrusor pressure. CONCLUSIONS: UCMSC transplantation can effectively improve neurological functional recovery after spinal cord injury, and its efficacy is superior to that of rehabilitation therapy and self-healing. TRIAL REGISTRATION: The present clinical study was registered at chictr.org (registration number: NCT01393977).

Automatic foreground extraction based on difference of Gaussian.

A novel algorithm for automatic foreground extraction based on difference of Gaussian (DoG) is presented. In our algorithm, DoG is employed to find the candidate keypoints of an input image in different color layers. Then, a keypoints filter algorithm is proposed to get the keypoints by removing the pseudo-keypoints and rebuilding the important keypoints. Finally, Normalized cut (Ncut) is used to segment an image into several regions and locate the foreground with the number of keypoints in each region. Experiments on the given image data set demonstrate the effectiveness of our algorithm.

Spatial patterns of whole brain grey and white matter injury in patients with occult spastic diplegic cerebral palsy.

Spastic diplegic cerebral palsy (SDCP) is a common type of cerebral palsy (CP), which presents as a group of motor-impairment syndromes. Previous conventional MRI studies have reported abnormal structural changes in SDCP, such as periventricular leucomalacia. However, there are roughly 27.8% SDCP patients presenting normal appearance in conventional MRI, which were considered as occult SDCP. In this study, sixteen patients with occult SDCP and 16 age- and sex-matched healthy control subjects were collected and the data were acquired on a 3T MR system. We applied voxel-based morphometry (VBM) and tract-based spatial statistics (TBSS) analysis to investigate whole brain grey and white matter injury in occult SDCP. By using VBM method, the grey matter volume reduction was revealed in the bilateral basal ganglia regions, thalamus, insula, and left cerebral peduncle, whereas the white matter atrophy was found to be located in the posterior part of corpus callosum and right posterior corona radiata in the occult SDCP patients. By using TBSS, reduced fractional anisotropy (FA) values were detected in multiple white matter regions, including bilateral white matter tracts in prefrontal lobe, temporal lobe, internal and external capsule, corpus callosum, cingulum, thalamus, brainstem and cerebellum. Additionally, several regions of white matter tracts injury were found to be significantly correlated with motor dysfunction. These results collectively revealed the spatial patterns of whole brain grey and white matter injury in occult SDCP.

A preliminary evaluation of efficacy and safety of Wharton's jelly mesenchymal stem cell transplantation in patients with type 2 diabetes mellitus.

INTRODUCTION: Stem cell therapy has recently been introduced to treat patients with type 2 diabetes mellitus (T2DM). However, no data are available on the efficacy and safety of allogeneic Wharton's Jelly-derived mesenchymal stem cell (WJ-MSC) transplantation in patients with T2DM. Here we performed a non-placebo controlled prospective phase I/II study to determine efficacy and safety of WJ-MSC transplantation in T2DM. METHODS: Twenty-two patients with T2DM were enrolled and received WJ-MSC transplantation through one intravenous injection and one intrapancreatic endovascular injection (catheterization). They were followed up for 12 months after transplantation. The primary endpoints were changes in the levels of glycated hemoglobin and C-peptide and the secondary endpoints included insulin dosage, fasting blood glucose (FBG), post-meal blood glucose (PBG), inflammatory markers and T lymphocyte counts. RESULTS: WJ-MSC transplantation significantly decreased the levels of glucose and glycated hemoglobin, improved C-peptide levels and beta cell function, and reduced markers of systemic inflammation and T lymphocyte counts. No major WJ-MSC transplantation-related adverse events occurred, but data suggest a temporary decrease in levels of C-peptide and beta cell function at one month after treatment, possibly related to intrapancreatic endovascular injection. CONCLUSIONS: Our data demonstrate that treatment with WJ-MSCs can improve metabolic control and beta cell function in patients with T2DM. The therapeutic mechanism may involve improvements in systemic inflammation and/or immunological regulation. TRIAL REGISTRATION: Chinese Clinical Trial Register ChiCTR-ONC-10000985. Registered 23 September 2010.

Comparative analysis of curative effect of CT-guided stem cell transplantation and open surgical transplantation for sequelae of spinal cord injury.

BACKGROUND: This study compared the clinical efficacies, advantages and disadvantages of two transplantation approaches for treating spinal cord injury: open surgical exploration combined with local stem cell transplantation (referred to as open surgical transplantation) and local stem cell transplantation by CT-guided puncture (referred to as CT-guided transplantation). METHODS: The patients were divided into the following three groups to perform a retrospective controlled study: Group A included nine patients who underwent open surgical transplantation, Group B included nine patients who underwent CT-guided transplantation, and Group C included nine patients who did not receive stem cell transplantation. The Abbreviated Injury Scale (AIS), the American Spinal Injury Association (ASIA) score and the motor evoked potentials (MEP) examination were utilized to compare the differences in the clinical efficacies. The advantages and disadvantages of the two transplantation approaches were also compared, including the surgical risks, the possibility of repeating the operation, the interval between surgery and rehabilitation exercises and the scope of conditions suitable for the operation. RESULTS: Whether evaluated by the AIS grading scale, the ASIA score or the MEP results, there were significant differences in the clinical efficacy among the three patient groups. Group B exhibited the best clinical outcome, followed by Group A, and Group C fared the worst. The CT-guided transplantation had the advantages of lower surgical risk, the potential to repeat the operations within a short time-frame and a short interval between surgery and rehabilitation exercise compared with the open surgical transplantation. The conditions that are suitable for CT-guided transplantation versus the conditions suitable for open surgical transplantation are not identical. The application scopes for the two approaches had their respective strengths. CONCLUSIONS: CT-guided stem cell transplantation was confirmed as a safe and effective approach to treat sequelae of spinal cord injury with the advantages of simpler operation, minimal invasion, less adverse reaction and quicker recovery. CLINICAL TRIALS REGISTRATION NUMBER: ChiCTR-TNRC-12002477.

Effects of bone marrow mesenchymal stromal cells on gross motor function measure scores of children with cerebral palsy: a preliminary clinical study.

BACKGROUND AIMS: Pre-clinical evidence indicates that autologous bone marrow-derived mesenchymal stromal cell (BM-MSC) transplantation improves motor function in patients with central nervous system disorders. METHODS: After providing informed consent, 52 patients with cerebral palsy (CP) who met the study criteria received BM-MSC transplantation. Gross motor function was assessed using the Gross Motor Function Measure (GMFM)-88 and GMFM-66 scales at baseline (before transplantation) and at 1 month, 6 months and 18 months post-transplantation. The participants completed the trial without visible side effects. The GMFM-66 percentile (motor growth curves) was used as the control index of motor function to exclude the interference of improvement with age. RESULTS: The score domains A, B, C and D and the total GMFM-88 and GMFM-66 scores in participants increased at 1 month, 6 months and 18 months post-transplantation compared with the baseline value (P < 0.01). The scores of domain E also increased at 6 months and 18 months post-transplantation, although they were not significantly increased at 1 month post-transplantation. There were significant increases in the GMFM-66 score and the GMFM-66 percentile corresponding to patient age and Gross Motor Function Classification System level after cell transplantation. CONCLUSIONS: Autologous BM-MSC transplantation appears to be a feasible, safe and effective therapy for patients with CP. The treatment improved the development of children with CP with regard to motor function.

Transplantation of autologous bone marrow mesenchymal stem cells in the treatment of complete and chronic cervical spinal cord injury.

Neuronal injuries have been a challenging problem for treatment, especially in the case of complete and chronic cervical spinal cord injury (SCI). Recently, particular attention is paid to the potential of stem cell in treating SCI, but there are only few clinical studies and insufficient data. This study explored the efficacy of autologous bone marrow mesenchymal stem cells (BMMSCs) transplantation in the treatment of SCI. Forty patients with complete and chronic cervical SCI were selected and randomly assigned to one of the two experimental groups, treatment group and control group. The treatment group received BMMSCs transplantation to the area surrounding injury, while the control group was not treated with any cell transplantation. Both the transplant recipients and the control group were followed up to 6 months, postoperatively. Preoperative and postoperative neurological functions were evaluated with AIS grading, ASIA score, residual urine volume and neurophysiological examination. Results showed that in the treatment group 10 patients had a significant clinical improvement in terms of motor, light touch, pin prick sensory and residual urine volume, while nine patients showed changes in AIS grade. Neurophysiological examination was consistent with clinical observations. No sign of tumor was evident until 6 months postoperatively. In the control group, no improvement was observed in any of the neurological functions specified above. BMMSCs transplantation improves neurological function in patients with complete and chronic cervical SCI, providing valuable information on applications of BMMSCs for the treatment of SCI.

Umbilical cord mesenchymal stem cell transplantation significantly improves neurological function in patients with sequelae of traumatic brain injury.

The aim of this study was to investigate the effects of transplantation with umbilical cord mesenchymal stem cells in patients with sequelae of traumatic brain injury (TBI). The study hypothesis was that umbilical cord mesenchymal stem cell transplantation could safely and effectively improve neurological function in patients with sequelae of traumatic brain injury. Forty patients with sequelae of TBI were randomly assigned to the stem cell treatment group or the control group. The patients in the stem cell treatment group underwent 4 stem cell transplantations via lumbar puncture. All patients of the group were also evaluated using Fugl-Meyer Assessments (FMA) and Functional Independence Measures (FIM) before and at 6 months after the stem cell transplantation. The patients in the control group did not receive any medical treatment (i.e., neither surgery nor medical intervention), and their FMA and FIM scores were determined on the day of the visit to the clinic and at 6 months after that clinical observation. The FMA results demonstrated an improvement in upper extremity motor sub-score, lower extremity motor sub-score, sensation sub-score and balance sub-score in the stem cell transplantation group at 6 months after the transplantation (P<0.05). The FIM results also exhibited significant improvement (P<0.05) in the patient self-care sub-score, sphincter control sub-score, mobility sub-score, locomotion sub-score, communication sub-score and social cognition sub-score. The control group exhibited no improvements after 6 months (P>0.05). All in all, the study results confirmed that the umbilical cord mesenchymal stem cell transplantation improved the neurological function and self-care in patients with TBI sequels. Umbilical cord mesenchymal stem cell transplantation may be a potential treatment for patients with sequelae of TBI. Further research, including a multicenter and large sample size prospective randomized clinical trial, will be required to define definitively the role of umbilical cord mesenchymal stem cell transplantation on sequelae of TBI.

Treatment of one case of cerebral palsy combined with posterior visual pathway injury using autologous bone marrow mesenchymal stem cells.

BACKGROUND: Cerebral palsy is currently one of the major diseases that cause severe paralysis of the nervous system in children; approximately 9-30% of cerebral palsy patients are also visually impaired, for which no effective treatment is available. Bone marrow mesenchymal stem cells (BMSCs) have very strong self-renewal, proliferation, and pluripotent differentiation potentials. Therefore, autologous BMSC transplantation has become a novel method for treating cerebral palsy. METHODS: An 11-year-old boy had a clear history of dystocia and asphyxia after birth; at the age of 6 months, the family members observed that his gaze roamed and noted that he displayed a lack of attention. A brain MRI examination at the age of 7 years showed that the child had cerebral palsy with visual impairment (i.e., posterior visual pathway injury). The patient was hospitalized for 20 days and was given four infusions of intravenous autologous BMSCs. Before transplantation and 1, 6, and 12 months after transplantation, a visual evoked potential test, an electrocardiogram, routine blood tests, and liver and kidney function tests were performed. RESULTS: The patient did not have any adverse reactions during hospitalization or postoperative follow-up. After discharge, the patient could walk more smoothly than he could before transplantation; furthermore, his vision significantly improved 6 months after transplantation, which was also supported by the electrophysiological examinations. CONCLUSIONS: The clinical application of BMSCs is effective for improving vision in a patient with cerebral palsy combined with visual impairment.

Antifungal activity of plant extracts against Colletotrichum lagenarium, the causal agent of anthracnose in cucumber.

BACKGROUND: Colletotrichum lagenarium is an important plant-pathogenic fungus that causes anthracnose of cucumber, a disease that is widespread under both greenhouse and field cultivation. To find a promising method for the control of this disease, extracts of eight plants from China were screened in the study presented here. RESULTS: The results showed that the extract of Cinnamomum camphora (L.) Presl had significantly greater inhibitory activity against C. lagenarium than the other seven plant extracts tested in vitro. At 5 days after inoculation in a potted seedling experiment, C. camphora extract demonstrated 95% control of C. lagenarium at a concentration of 16 mg mL⁻¹, and the MIC50 was 2.596 mg mL⁻¹. Microscopic observation showed that this extract had a significant impact on the morphology of the fungus, leading to shorter hyphae that were more branched; this observation was associated with the inhibition of mycelial growth. Stability assays revealed that the extract was relatively stable at 80 °C, under acidic conditions and when exposed to light and short periods of UV radiation. CONCLUSION: These results demonstrated that the extract from C. camphora could be used as a potent phytochemical fungicide.

Triterpenoids and steroids from the fruits of Melia toosendan and their cytotoxic effects on two human cancer cell lines.

Ten new triterpenoids, named meliasenins I-R (1-10), one new steroid (11), and 11 related known compounds (12-22) were isolated from fruits of Melia toosendan. The structures of the new compounds were established on the basis of spectroscopic methods, including 2D NMR techniques and mass spectrometry. The relative configuration of 1, (20R*,23E)-25-hydroperoxyeupha-7,23-diene-3ß,16ß-diol (meliasenin I), was confirmed by single-crystal X-ray diffraction analysis. All isolated triterpenoids (1-10, 12-15) and two steroids (11, 20) were tested for their cytotoxicity against U20S human osteosarcoma and MCF-7 human breast cancer cells using the MTT assay, and some of them were significantly cytotoxic (IC(50) <10 µg/mL). The insecticidal properties of compounds 1-15 and 20 were also briefly evaluated.

[Sinerem labeling and MRI tracking of neural stem cells in vivo and in vitro].

OBJECTIVE: To label rat neural stem cells (NSCs) with the complex of Sinerem, the ultrasmall superparamagnetic iron oxide (USPIO), and poly-L-lysine (PLL), and evaluate the feasibility of tracking the labeled cells with magnetic resonance imaging (MRI) in vitro and in vivo. METHODS: Sinerem was incubated with PLL to obtain the complex of Sinerem-PLL. The mesenchymal stem cells (MSCs) isolated from the bone marrow of SD rats were cultured and induced to differentiate into the neural stem cells. The second-passage cells were cultured overnight with the Sinerem-PLL complex, after which Prussian blue staining and transmission electron microscopy were performed to observe the nanoparticles in the cytoplasm. Cell apoptosis assay was performed to assess the cell viability 1 day, 1 week, and 2 weeks after the labeling. Cell tracking with 4.7 MR system was carried out in vivo and in vitro using T(2)WI and T(2)*WI sequences. RESULTS: The NSCs could be effectively labeled with Sinerem-PLL complex with the labeling efficiency exceeding 95%. Prussian blue staining showed numerous blue iron particles in the cytoplasm, and under transmission electron microscope, these particles accumulated in the endosomes/lysosomes. The labeling did not significantly affect the cell viability and proliferation. Remarkable low signal density changes of the labeled cells was seen on T(2)WI and T(2)*WI in vivo and in vitro. CONCLUSION: NSCs can be effectively labeled with Sinerem-PLL complex, and MRI can be used to track the labeled cells in vivo and in vitro.