[Pilot study on comprehensive malaria control in Judong Village, Congjiang County, Guizhou Province].

OBJECTIVE: To explore the working mode of continuous malaria elimination. METHODS: Judong Village in Congjiang County, Guizhou Province, which had the highest record of malaria incidence in the history of Guizhou Province, was selected as a pilot, a comprehensive prevention and control intervention was carried out aiming at the malaria epidemic, transmission and influencing factors, and the effect was evaluated after the intervention. RESULTS: From 2015 to 2016, through those intensive measures of constructing system, improving environment and health education, the people's awareness of malaria prevention was increased, the people's enthusiasm and initiatives to participate in the prevention and control of malaria were improved in Judong Village. Meanwhile, the roads were hardened and beautified, the dispose of sewage, feces and garbage was centralized. The awareness rate of malaria prevention knowledge of residents increased to 85% to 95%, and the formation rate of anti-mosquito behavior increased to 89.66%. The density of media Anopheles reduced from 0.56 to 0.07 mosquitoes per hour each mosquito catching platform. CONCLUSIONS: A long-term mechanism for malaria prevention and control has been almost constructed in Judong Village, which facilitates the consolidation of malaria control achievements, and the progress towards malaria elimination.

The chemokine CXCL1 and its receptor CXCR2 contribute to chronic stress-induced depression in mice.

Depression is increasingly recognized as an inflammatory disease, with inflammatory crosstalk in the brain contributing its pathogenesis. Life stresses may up-regulate inflammatory processes and promote depression. Although cytokines are central to stress-related immune responses, their contribution to stress-induced depression remains unclear. Here, we used unpredictable chronic mild stress (UCMS) to induce depression-like behaviors in mice, as assessed through a suite of behavioral tests. C-X-C motif chemokine ligand 1 (CXCL1)-related molecular networks responsible for depression-like behaviors were assessed through intrahippocampal microinjection of lenti-CXCL1, the antidepressant fluoxetine, the C-X-C motif chemokine receptor 2 (CXCR2) inhibitor SB265610, and the glycogen synthase kinase-3ß (GSK3ß) inhibitor AR-A014418. Modulation of apoptosis-related pathways and neuronal plasticity were assessed via quantification of cleaved caspase-3, B-cell lymphoma 2-associated X protein, cAMP response element-binding protein (CREB), and brain-derived neurotrophic factor (BDNF) protein expression. CXCL1/CXCL2 expression was correlated with depression-like behaviors in response to chronic stress or antidepressant treatment in the UCMS depression model. Intrahippocampal microinjection of lenti-CXCL1 increased depression-like behaviors, activated GSK3ß, increased apoptosis pathways, suppressed CREB activation, and decreased BDNF. Administration of the selective GSK3ß inhibitor AR-A014418 abolished the effects of lenti-CXCL1, and the CXCR2 inhibitor SB265610 prevented chronic stress-induced depression-like behaviors, inhibited GSK3ß activity, blocked apoptosis pathways, and restored BDNF expression. The CXCL1/CXCR2 axis appears to play a critical role in stress-induced depression, and CXCR2 is a potential novel therapeutic target for patients with depression.-Chai, H.-H., Fu, X.-C., Ma, L., Sun, H.-T., Chen, G.-Z., Song, M.-Y., Chen, W.-X., Chen, Y.-S., Tan, M.-X., Guo, Y.-W., Li, S.-P. The chemokine CXCL1 and its receptor CXCR2 contribute to chronic stress-induced depression in mice.

Transplantation of adipocyte-derived stem cells in a hydrogel scaffold for the repair of cortical contusion injury in rats.

Adipocyte-derived stem cells have emerged as a novel source of stem cell therapy for their autologous and readily accessible and pluripotent potential to differentiate into different lineages such as neural stem cells (NSCs) and endothelial progenitor cells (EPCs). Transplantation of NSCs and EPCs has been promising for the repair of brain injury. We explored using co-transplanted hydrogel scaffold to improve the survival of the transplanted cells and recovery of neurological function. Adult Wistar rats were transplanted with EPC-hydrogel, NSC-hydrogel, NSC-EPC-hydrogel, EPC only, or NSC only 7 days after cortical contusion injury. Behavioral tests were performed to evaluate neurological function before, and 1, 2, 3, and 4 weeks after transplantation. Size of injury, extent of vascularization, as well as the survival and differentiation of the transplanted EPCs and NSCs, were evaluated at week 5. All transplantation groups displayed significantly better neurological function compared with the control groups. Improved neurological function correlated with significantly smaller injury volumes than that of the saline group. Using immunostaining, we have shown that while transplanted NSCs differentiated into both neurons and astrocytes, the EPCs were incorporated into vessel epithelia. The extent of reactive gliosis (based on glial fibrillary acidic protein immunostaining) was significantly reduced in all treatment groups (NSC-EPC-hydrogel, NSC-hydrogel, and EPC-hydrogel) when compared with the saline group, with the highest reduction in the NSC-EPC-hydrogel transplantation group. Thus, co-transplantation of hydrogel scaffold provides a more conducive environment for the survival and differentiation of NSCs and EPCs at the site of brain injury, leading to improved vascularization and better recovery of neurological function.

IDN2 and its paralogs form a complex required for RNA-directed DNA methylation.

IDN2/RDM12 has been previously identified as a component of the RNA-directed DNA methylation (RdDM) machinery in Arabidopsis thaliana, but how it functions in RdDM remains unknown. By affinity purification of IDN2, we co-purified two IDN2 paralogs IDP1 and IDP2 (IDN2 PARALOG 1 and 2). The coiled-coil domain between the XS and XH domains of IDN2 is essential for IDN2 homodimerization, whereas the IDN2 C-terminal XH domain but not the coiled-coil domain is required for IDN2 interaction with IDP1 and IDP2. By introducing the wild-type IDN2 sequence and its mutated derivatives into the idn2 mutant for complementation testing, we demonstrated that the previously uncharacterized IDN2 XH domain is required for the IDN2-IDP1/IDP2 complex formation as well as for IDN2 function. IDP1 is required for de novo DNA methylation, siRNA accumulation, and transcriptional gene silencing, whereas IDP2 has partially overlapping roles with IDP1. Unlike IDN2, IDP1 and IDP2 are incapable of binding double-stranded RNA, suggesting that the roles of IDP1 and IDP2 are different from those of IDN2 in the IDN2-IDP1/IDP2 complex and that IDP1 and IDP2 are essential for the functioning of the complex in RdDM.

Artemisinin biosynthesis enhancement in transgenic Artemisia annua plants by downregulation of the ß-caryophyllene synthase gene.

Artemisinin is an effective antimalarial drug isolated from the medicinal plant Artemisia annua L. Due to its increasing market demand and the low yield in A. annua, there is a great interest in increasing its production. In this paper, in an attempt to increase artemisinin content of A. ANNUA by suppressing the expression of ß-caryophyllene synthase, a sesquiterpene synthase competing as a precursor of artemisinin, the antisense fragment (750 bp) of ß-caryophyllene synthase cDNA was inserted into the plant expression vector pBI121 and introduced into A. annua by Agrobacterium-mediated transformation. PCR and Southern hybridization confirmed the stable integration of multiple copies of the transgene in 5 different transgenic lines of A. annua. Reverse transcription PCR showed that the expression of endogenous CPS in the transgenic lines was significantly lower than that in the wild-type control A. annua plants, and ß-caryophyllene content decreased sharply in the transgenic lines in comparison to the control. The artemisinin content of one of the transgenic lines showed an increase of 54.9 % compared with the wild-type control. The present study demonstrated that the inhibition pathway in the precursor competition for artemisinin biosynthesis by anti-sense technology is an effective means of increasing the artemisinin content of A. annua plants.

Human umbilical vein-derived dopaminergic-like cell transplantation with nerve growth factor ameliorates motor dysfunction in a rat model of Parkinson's disease.

Mesenchymal stem cells are capable of differentiating into dopaminergic-like cells, but currently no report has been available to describe the induction of human umbilical vein mesenchymal stem cells (HUVMSCs) into dopaminergic-like cells. In this study, we induced HUVMSCs in vitro into neurospheres constituted by neural stem-like cells, and further into cells bearing strong morphological, phenotypic and functional resemblances with dopaminergic-like cells. These HUVMSC-derived dopaminergic-like cells, after grafting into the brain of a rat model of Parkinson's disease (PD), showed a partial therapeutic effect in terms of the behavioral improvement. Nerve growth factor was reported to improve the local microenvironment of the grafted cells, and we therefore further tested the effect of dopaminergic-like cell grafting combined with nerve growth factor (NGF) administration at the site of cell transplantation. The results showed that NGF administration significantly promoted the survival of the grafted cells in the host brain and enhanced the content of dopaminergic in the local brain tissue. Behavioral test demonstrated a significant improvement of the motor function of the PD rats after dopaminergic-like cell grafting with NGF administration as compared with that of rats receiving the cell grafting only. These results suggest that transplantation of the dopaminergic-like cells combined with NGF administration may represent a new strategy of stem cell therapy for PD.

[Construction of a mouse pDsRed2-N1-SDF-1alpha eukaryotic expression vector and its expression in mouse bone marrow mesenchymal stem cells].

OBJECTIVE: To construct the eukaryotic expression vector pDsRed2-N1-SDF-1alpha and observe its expression in the mouse bone marrow mesenchymal stem cells. METHOD: SDF-1alpha gene sequence with XhoI, EcoRI restriction enzyme cutting site was amplified from the total RNA of mouse smooth muscle cells by reverse transcription-polymerase chain reaction (RT-PCR) and inserted into the eukaryotic expression vector pDsRed2-N1 encoding red fluorescent protein gene, and the insertion was verified by endonuclease digestion and DNA sequencing. Mouse bone marrow mesenchymal stem cells identified with immunofluorescence assay for vimentin expression were transfected with the constructed plasmid pDsRed2-N1-SDF-1alpha, and the expression of sdf-1alpha was detected using immunofluorescence assay. RESULTS: The DNA fragment amplified by PCR from the total RNA was identical to SDF-1alpha from the gene library, and an identical DNA fragment was also amplified from the recombinants. Sequence analysis confirmed the successful insertion of SDF-1alpha into the pDsRed2-N1 vector and the eukaryotic expression vector pDsRed2-N1-SDF-1alpha was successfully constructed. The cultured mouse bone marrow mesenchymal stem cells positive for vimentin protein showed SDF-1alpha expression 24 h after transfection with the recombinant vector. CONCLUSION: The pDsRed2-N1-SDF-1alpha eukaryotic expression vector constructed is capable of expression of SDF-1alpha fusion protein in the mouse bone marrow mesenchymal stem cells.

Identification of a Polygonum cuspidatum three-intron gene encoding a type III polyketide synthase producing both naringenin and p-hydroxybenzalacetone.

Benzalacetone synthase (BAS) is a member of the plant-specific type III PKS superfamily that catalyzes a one-step decarboxylative condensation of 4-coumaroyl-CoA with malonyl-CoA to produce p-hydroxybenzalacetone. In our recent work (Ma et al. in Planta 229(3):457-469, 2008), a three-intron type III PKS gene (PcPKS2) was isolated from Polygonum cuspidatum Sieb. et Zucc. Phylogenetic and functional analyses revealed this recombinant PcPKS2 to be a BAS. In this study, another three-intron type III PKS gene (PcPKS1) and its corresponding cDNA were isolated from P. cuspidatum. Sequence and phylogenetic analyses demonstrated that PcPKS1 is a chalcone sythase (CHS). However, functional and enzymatic analyses showed that recombinant PcPKS1 is a bifunctional enzyme with both, CHS and BAS activity. DNA gel blot analysis indicated that there are two to four CHS copies in the P. cuspidatum genome. RNA gel blot analysis revealed that PcPKS1 is highly expressed in the rhizomes and in young leaves, but not in the roots of the plant. PcPKS1 transcripts in leaves were inducible by pathogen infection and wounding. BAS is thought to play a crucial role in the construction of the C(6)-C(4) moiety found in a variety of phenylbutanoids, yet so far phenylbutanoids have not been isolated from P. cuspidatum. However, since PcPKS1 and PcPKS2 (Ma et al. in Planta 229(3):457-469, 2008) have been identified in P. cuspidatum, it is possible that such compounds are also produced in that plant, albeit in low concentrations.

[Culture and identification of neural stem cells derived from the subventricular zone of adult mice].

OBJECTIVE: To establish a method for culturing and identifying neural stem cells (NSCs) derived from the subventricular zone (SVZ) in adult mice. METHODS: NSCs were isolated from the SVZ of adult mouse brain and cultured in serum-free medium. Cell cloning and BrdU incorporation were performed to identify the self-renewal and proliferative capacity of the NSCs. Fluorescence immunocytochemistry was used to examine the expressions of the NSC markers nestin and SOX2, neuronal marker Tuj1, astrocyte marker GFAP and oligodendrocyte marker NG2. The expressions of nestin and SOX2 were further examined by Western blotting and RT-PCR. RESULTS: NSCs with self-renewal and proliferative capacity were obtained from the SVZ of adult mice and grown as floating neurospheres. The NSCs expressed nestin and SOX2 and could differentiated into Tuj1-positive neurons, GFAP-positive astrocytes and NG2-positive oligodendrocytes. CONCLUSION: This method allows simple and stable culture of NSCs from the SVZ of adult mice.

[Application of virtual imaging technique in diagnosis of intracranial aneurysms].

OBJECTIVE: To investigate the value of virtual imaging technique in diagnosis of intracranial aneurysms. METHODS: Fifty-four cases of 54 intracranial aneurysm diagnosed by three-dimensional CT angiography (3D-CTA) examinations were enrolled in this study. Three-dimensional virtual images of the skull and cerebral vessels were acquired by three-dimensional reconstruction of the original CT images using the surgical planning system, and the location, size and shape of the aneurysms and their anatomical relationship with the adjacent tissues were observed and measured from several angles. All the patients underwent surgical planning and simulated surgical operations using the virtual surgical instruments available in the system. RESULTS: All the 54 cases had successful three-dimensional virtual image reconstruction and the surgical planning operations. The virtual imaging system generated clear and vivid three-dimensional virtual images which clearly visualized the location and size of the aneurysms and their precise anatomical relations to the parent arteries and skull. This virtual reality imaging system also allowed simulation of simple surgical procedures. CONCLUSION: The surgical planning system based on the virtual reality imaging can serve as a useful means to assist the diagnosis and provide precise imaging details of intracranial aneurysms.

Combined application of virtual imaging techniques and three-dimensional computed tomographic angiography in diagnosing intracranial aneurysms.

BACKGROUND: The diagnostic value of virtual imaging combined with three-dimensional computed tomographic angiography (3D-CTA) for intracranial aneurysms has not been fully elucidated yet. This study aimed to evaluate the value of combined application of virtual imaging techniques and 3D-CTA in diagnosing patients with aneurismal subarachnoid hemorrhage (SAH) at the acute stage. METHODS: Eighty patients with non-traumatic SAH received 3D-CTA examinations. The raw CT data of these patients were reconstructed and transferred into the 3D mode through the surgical plan system based on virtual reality (VR) image, and the 3D virtual images of skulls and brain blood vessels were acquired. The location, size and shape of aneurysms and their anatomic relationship with adjacent tissues were measured from many points of view. RESULTS: Seventy-three aneurysms were detected in 68 of the 80 patients, but 2 aneurysms were detected in 2 of the 5 patients who had been found free of aneurysms previously and had received 3D-CTA examinations for a second time one month later. The 3D virtual images produced by the virtual imaging system were clear and vivid, and they could reveal the location and size of the aneurysm and its relations to the parent artery and skull directly. CONCLUSIONS: The imaging of 3D-CTA is convenient, reliable and fast in diagnosing intracranial aneurysms and can be regarded as the first choice for the diagnosis and treatment of ruptured intracranial aneurysms. Combined with the surgical plan system based on the VR image, 3D-CTA may obtain more imaging information about aneurysms.

[Time course of calpain activity changes in rat neurons following fluid percussion injury and the interventional effect of mild hypothermia].

OBJECTIVE: To investigate the time course of calpain activity changes in rat neurons following fluid percussion injury (FPI) under normothermia (37 degrees celsius;) and mild hypothermia (32-/+0.5) degrees celsius;. METHODS: In vitro cultured rat neurons were subjected to FPI followed by application of mild hypothermia for intervention at different time points, and the changes in intraneuronal calpain activity following FPI and the interventional effect of mild hypothermia on calpain activity were evaluated by UV-spectrophotometry at different time points. RESULTS: Remarkable changes occurred in calpain activity in the neurons following FPI at 37 degrees celsius;, and mild hypothermia produced obvious interventional effect on calpain activity in close relation to the timing of intervention initiation. CONCLUSION: Intraneuronal calpain activity changes following FPI are involved in the pathological process of cellular injury, and mild hypothermia might offer protection against traumatic brain injury to some extent by regulating calpain activity. The interventional effect of mild hypothermia is associated with the timing of the intervention initiation.

[Detection of amino acid neurotransmitters in cerebrospinal fluid of patients in persistent vegetative state].

OBJECTIVE: To detect the amino acid neurotransmitters in cerebrospinal fluid of patients in persistent vegetative state (PVS) to define their relation to the clinical manifestations of the patients. METHODS: The cerebrospinal fluid from 46 patients in PVS and 20 control patients were collected for detection of glutamic acid, aspartic acid, alanine and glycine by high-performance liquid chromatography. RESULTS: All the four amino acids were separated by chromatography within 25 min. The contents of all the 4 neurotransmitters in PVS group were higher than those in the control group, and their contents were inversely correlated to PVS score of the patients. The differences in the concentration of glycine and alanine were significant between PVS patients and the control patients (P<0.05). CONCLUSION: The patient's clinical manifestations are related to the contents of amino acid neurotransmitters in the cerebrospinal fluid of PVS patients.

[Pathological changes of cat brain tissues after cranial gunshot wound in hot and humid environment].

OBJECTIVE: To investigate pathological changes in cat brain tissues after gunshot wound in the head in hot and humid environments. METHODS: Sixteen cats were randomly divided into 4 groups, namely the normal environment (group 1), gunshot in normal environment (group 2), hot and humid environment (group 3), and gunshot wound in hot and humid environment (group 4) groups. Pathological changes of the cat brain tissues were observed with both optical and electron microscopes. RESULTS: The early-stage changes in group 2 were high-lighted by vasomotor dysfunction, with the coexistence of both vascular spasm and dilation. In group 3, vascular spasm was depressed while hemorrhagic changes increased. On the brain tissue sections in group 4 for optical microscopic observation and ultra-thin sections for electron microscopic examination, the number of viable neural cells was obviously reduced, and edema, degeneration of the organelle occurred; loosening of the capillary tight junction, rupture and bleeding of the blood vessels, as well as degeneration and loosening of the myelin sheath were observed by transmission and scanning electron microscopes, and such changes were more serious than those in group 2. CONCLUSION: The hot and humid environment can significantly affect the pathological changes in the brain tissues of cat with cranial gunshot wound.

Bacterial growth characteristics in craniocerebral gunshot wound of cat in a hot and humid environment.

OBJECTIVE: To investigate the growth behavior of characteristics in craniocerebral gunshot wound of cats in a hot and humid environment. METHODS: Twenty-three cross-bred cats were randomly divided into 4 groups: group A, the gunshot wound control group at normal temperature, in which tissue sampling was performed immediately after the wounding; group B, another gunshot wound control group at normal temperature, in which the samples were taken 6 h after the wounding; group C, the gunshot wound group subjected to a hot and humid environment, in which the tissue samples were obtained 6 h after the wounding; group D, the control group without undergoing the wound. The tissues from the wound tract and the surrounding tissues were sampled for bacterial culture and counting. RESULTS: The bacterial counts of the tissues from the wound tract, the tissues within 5 mm and within 5-10 mm from wound tract varied insignificantly between groups A, B and C (P>0.05). In each group, the bacterial counts declined in the tissues as the distance of the sampling sites from the wound tract increased (P<0.01). The bacterial counts of the tissues from the wound tract and within 5 mm from the wound tract in group A, B and C were significantly different from those in group D (P<0.01). CONCLUSION: Hot and humid environment does not significantly affect the bacterial growth in the craniocerebral gunshot wound within the first 6 h, which is a safe period against rapid bacterial growth and suitable for debridement.