Applications and Prospects of CRISPR/Cas9-Mediated Base Editing in Plant Breeding.

The clustered regularly interspaced short palindromic repeats (CRISPR)/associated protein 9 system (Cas9) has been used at length to optimize multiple aspects of germplasm resources. However, large-scale genomic research has indicated that novel variations in crop plants are attributed to single-nucleotide polymorphisms (SNPs). Therefore, substituting single bases into a plant genome may produce desirable traits. Gene editing by CRISPR/Cas9 techniques frequently results in insertions-deletions (indels). Base editing allows precise single-nucleotide changes in the genome in the absence of double-strand breaks (DSBs) and donor repair templates (DRTs). Therefore, BEs have provided a new way of thinking about genome editing, and base editing techniques are currently being utilized to edit the genomes of many different organisms. As traditional breeding techniques and modern molecular breeding technologies complement each other, various genome editing technologies have emerged. How to realize the greater potential of BE applications is the question we need to consider. Here, we explain various base editings such as CBEs, ABEs, and CGBEs. In addition, the latest applications of base editing technologies in agriculture are summarized, including crop yield, quality, disease, and herbicide resistance. Finally, the challenges and future prospects of base editing technologies are presented. The aim is to provide a comprehensive overview of the application of BE in crop breeding to further improve BE and make the most of its value.

Induction of cancer cell stemness in glioma through glycolysis and the long noncoding RNA HULC-activated FOXM1/AGR2/HIF-1α axis.

Gliomas are the most common primary intracranial tumor, accounting for more than 70% of brain malignancies. Studies indicate that highly upregulated in liver cancer (HULC), a long noncoding RNA (lncRNA), functions as an oncogene in gliomas. However, the underlying mechanism of HULC in gliomas remains under-studied and was subsequently investigated in the current study. Brain tissues were clinically collected from 50 patients with glioblastoma (GBM) and 35 patients with acute craniocerebral injury, followed by immunohistochemical detection of the expression patterns of Forkhead box M1 (FOXM1), anterior gradient 2 (AGR2), and hypoxia-inducible factor-1α (HIF-1α). After flow cytometry-based sorting of the CD133+ glioma stem cells (GSCs) from the U251 cell line, the obtained cells were subjected to lentivirus infection. Afterwards, the proliferation, stemness, and apoptosis of GSCs were evaluated using sphere formation, immunofluorescence, and flow cytometry assays, respectively. In addition, the interactions among HULC, FOXM1, AGR2, and HIF-1α were identified using RNA immunoprecipitation (RIP), RNA pull-down, Chromatin immunoprecipitation (ChIP), IP, and dual luciferase reporter assays. Last, the specific effects were validated in vivo. HULC was upregulated in GBM tissues and GSCs, which may promote the progression of glioma. On the other hand, silencing of HULC reduced the stemness, inhibited the proliferation, and promoted the apoptosis and differentiation of GSCs. In addition, HULC further stabilized FOXM1 expression in GSCs through ubiquitination, while FOXM1 activated AGR2 transcription to promote HIF-1α expression. Moreover, HULC promoted the glycolysis and stemness of GSCs through its regulation of the FOXM1/AGR2/HIF-1α axis, consequently exacerbating the occurrence and development of glioma. The findings obtained in our study indicate that HULC stabilizes the FOXM1 protein by ubiquitination to upregulate the expression of AGR2 and HIF-1α, which further promote the glycolysis of and maintain the stemness of GSCs, to enhance the tumorigenicity of GSCs, highlighting a novel therapeutic target for glioma.

Extracellular vesicles derived from M2 microglia reduce ischemic brain injury through microRNA-135a-5p/TXNIP/NLRP3 axis.

Accumulating evidences have suggested that extracellular vesicles (EVs) are crucial players in the pathogenesis of ischemic brain injury. This study was designed to explore the specific functions of M2 phenotype microglia-derived EVs in ischemic brain injury progression. The expression of microRNA-135a-5p (miR-135a-5p) in M2 microglia-derived EVs was determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), followed by the identification of expression relationship among miR-135a-5p, thioredoxin-interacting protein (TXNIP), and nod-like receptor protein 3 (NLRP3) by dual luciferase reporter gene assay. After construction of an oxygen-glucose deprivation/reperfusion (OGD/R) cell model, the effects of miR-135a-5p on the biological characteristics of HT-22 cells were assessed by cell counting kit 8 (CCK-8) assay and flow cytometry. Finally, a mouse model of transient middle cerebral artery occlusion (tMCAO) was established and cerebral infarction volume was determined by triphenyltetrazolium chloride (TTC) staining and the expression of IL-18 and IL-1ß in the brain tissue was determined by enzyme-linked immunosorbent assay (ELISA). We found that M2 microglia-derived EVs had high expression of miR-135a-5p, and that miR-135a-5p in M2 microglia-derived EVs negatively regulated the expression of NLRP3 via TXNIP. Overexpression of miR-135a-5p promoted the proliferation but inhibited the apoptosis of neuronal cells, and inhibited the expression of autophagy-related proteins. M2 microglia-derived EVs delivered miR-135a-5p into neuronal cells to inhibit TXNIP expression, which further inhibited the activation of NLRP3 inflammasome, thereby reducing neuronal autophagy and ischemic brain injury. Hence, M2 microglia-derived EVs are novel therapeutic targets for ischemic brain injury treatment.

Occurrence, bioaccumulation and toxicological effect of drugs of abuse in aquatic ecosystem: A review.

Drugs of abuse are a group of emerging contaminants. As the prevalence of manufacture and consumption, there is a growing global environmental burden and ecological risk from the continuous release of these contaminants into environment. The widespread occurrence of drugs of abuse in waste wasters and surface waters is due to the incomplete removal through traditional wastewater treatment plants in different regions around the world. Although their environmental concentrations are not very high, they can potentially influence the aquatic organisms and ecosystem function. This paper reviews the occurrence of drugs of abuse and their metabolites in waste waters and surface waters, their bioaccumulation in aquatic plants, fishes and benthic organisms and even top predators, and the toxicological effects such as genotoxic effect, cytotoxic effect and even behavioral effect on aquatic organisms. In summary, drugs of abuse occur widely in aquatic environment, and may exert adverse impact on aquatic organisms at molecular, cellular or individual level, and even on aquatic ecosystem. It necessitates the monitoring and risk assessment of these compounds on diverse aquatic organisms in the further study.

New Synthesis for the Monobactam Antibiotic-LYS228.

A new synthesis of LYS228, fitting for further process development for commercial manufacture, is described. The key features of this synthesis include development of new protocols for acylation reactions, application of an asymmetric hydrogenation via dynamic kinetic resolution, and a late-stage ring closure to form ß-lactam 1.

Application of Transition-Metal Catalysis, Biocatalysis, and Flow Chemistry as State-of-the-Art Technologies in the Synthesis of LCZ696.

LCZ696 is a novel treatment for patients suffering from heart failure that combines the two active pharmaceutical ingredients sacubitril and valsartan in a single chemical compound. While valsartan is an established drug substance, a new manufacturing process suitable for large-scale commercial production had to be developed for sacubitril. The use of chemocatalysis, biocatalysis, and flow chemistry as state-of-the-art technologies allowed to efficiently build up the structure of sacubitril and achieve the defined performance targets.

Phthalate esters contamination in vegetable-soil system of facility greenhouses in Jingmen, central China and the assessment of health risk.

Residual levels of six priority phthalate esters (PAEs) compounds in top soils and vegetables of 40 selected greenhouses in two typical facility vegetable greenhouse production areas of Jingmen (Hubei, central China) were determined. Total concentrations of six target PAEs (Σ6PAEs) in 72 vegetable and 40 soil samples ranged from 862 ± 209 to 4260 ± 385 µg kg-1 (dry weight, DW), and from 1122 ± 156 to 4134 ± 272 µg kg-1 (DW), respectively. Di-n-butyl phthalate and di-(2-ethylhexyl) phthalate (DEHP) are the two most frequently and massively detected in all samples. The highest Σ6PAEs appeared in leaf samples of Tuanlin, vegetable fruits of Zhongxiang and soils of Tuanlin. No carcinogenic risk was posed based on the results of health risk assessment, but non-carcinogenic risk of DEHP to children less than 6 years old in Tuanlin (all 24 sampling sites) and Zhongxiang (4/16 sampling sites) and people older than 6 years old in Tuanlin (19/24 sampling sites) were achieved from hazard quotient values. The contamination risk problem of PAEs in Tuanlin deserves greatest concern in Jingmen. Combined with the results of our former study, the health risks of target pollutants were clarified and the lack of survey data on PAE concentrations in facility vegetable greenhouses of central China was filled in. Due to high residuals and significant non-carcinogenic risk values, DEHP should be nominated as priority PAEs in China. Our study suggested better regulation for PAEs control in intensively managed greenhouses and references for revision of Chinese environmental standards.

Silencing microRNA-221/222 cluster suppresses glioblastoma angiogenesis by suppressor of cytokine signaling-3-dependent JAK/STAT pathway.

Angiogenesis is a major pathologic characteristic of glioblastoma, which is one aggressive primary brain tumor. MicroRNA-221/222 (miR-221/222) cluster has been previously reported to function importantly in malignant glioma biological process. The current study aims at evaluating the effects of miR-221/222 cluster on angiogenesis of glioblastoma cells. Microarray data were analyzed to select glioblastoma-associated differentially expressed genes, and dual-luciferase reporter assay was performed to assess targeting correlation between miR-221/222 cluster and suppressor of cytokine signaling-3 (SOCS3). Subsequently, the expression patterns of miR-221 and miR-222 in glioblastoma cells were identified. miR-221 and miR-222 were overexpressed or silenced in glioblastoma cells to identify the effect of miR-221/222 cluster in cell invasion, migration, proliferation, and angiogenesis. To define downstream pathway of miR-221/222 cluster or SOCS3 in glioblastoma, levels of Janus kinase (JAK)/signal transducers and activators of transcription (STAT) pathway-related proteins were assessed. Additionally, the functions of miR-221/222 on glioblastoma cell angiogenesis were measured in vivo with microvessel density assayed. miR-221 and miR-222 were expressed at a high level and SOCS3 was at a low level in glioblastoma. Downregulation of the miR-221/222 cluster diminished the invasion, migration, proliferation, and angiogenesis with reduced protein levels of matrix metalloproteinase-2 (MMP-2), MMP-9, and vascular endothelial growth factor in glioblastoma cells. Also, silencing miR-221/222 cluster reduced p-JAK2/JAK2 and p-STAT3/STAT3. Consistently, the inhibitory role of silencing miR-221/222 cluster on tumorigenesis of glioblastoma cells was confirmed in vivo. Collectively, the inhibition of miR-221/222 cluster could attenuate the glioblastoma angiogenesis through inactivation of the JAK/STAT pathway by upregulating SOCS3.

[Microsurgical treatment of multi-segment cervical intramedullary ependymomas].

OBJECTIVE: To explore the microsurgical skills of multi-segment cervical ependymomas and evaluate preoperative and postoperative neurological functions. METHODS: A total of 61 patients with multi-segment cervical ependymoma from January 2008 to July 2013 at our hospital were analyzed to summarize their clinical features, imaging findings, diagnosis, tumor resection and changes of neurological functions. RESULTS: Total (n = 58) and subtotal (n = 3) resections were performed. And postoperative nerve functions unchanged (n = 43) and aggravated (n = 18). There was no mortality. CONCLUSION: Proper planning and refined microsurgical techniques are essential in the treatment of multi-segment cervical ependymomas.

MXene-reinforced octahedral PtCu nanocages with boosted electrocatalytic performance towards endocrine disrupting pollutants sensing.

Rational tailoring of hollow and porous bimetallic structures with excellent electrocatalytic performance is appealing yet challenging. Further, combining independent bimetallic nanoparticles with flexible two-dimensional substrate by forming stable heterocomplex is still highly desired for electrocatalysis. Herein, hierarchical PtCu alloy octahedrons with hollow interiors and nanosheet-assembled nanoshells were synthesized by a facile and efficient chemical transformation strategy using Cu2O as sacrificial templates. Such octahedral PtCu nanocages displayed significantly enhanced electrocatalytic activity owing to their unique hollow and porous architectures which provided easy access for analytes to the catalyst surface. Thereafter, introduction of Ti3C2Tx MXene was realized via simple incubation of Ti3C2Tx in solution containing the 3-aminopropyltriethoxysilane-capped PtCu, and their electrostatic interaction guaranteed the firm adsorption of PtCu nanocages on Ti3C2Tx nanosheets. It turned out that the sensitivity of the hybrid sensor was remarkably improved for electrochemical monitoring of endocrine disrupting pollutants in water, exhibiting ultrawide linear ranges and sub-nanomole detection limits. The eminent electrode performance is attributed to the high specific area, fast electrochemical kinetics, decent electrical catalytic ability, and the synergistic effect between Pt, Cu, and MXene.

Monitoring of trace aquatic sulfonamides through hollow zinc-nitrogen-carbon electrocatalysts anchored on MXene architectures.

Herein, we designed and fabricated hollow N-doped carbon polyhedrons with atomically dispersed Zn species (Zn@HNCPs) through a topo-conversion strategy by utilising metal-organic frameworks as precursors. Zn@HNCPs achieved efficient electrocatalytic oxidation of sulfaguanidine (SG) and phthalyl sulfacetamide (PSA) sulfonamides through the high intrinsic catalytic activity of the Zn-N4 sites and excellent diffusion from the hollow porous nanostructures. The combination of the novel Zn@HNCPs with two-dimensional Ti3C2Tx MXene nanosheets resulted in improved synergistic electrocatalytic performance for the simultaneous monitoring of SG and PSA. Therefore, the detection limit of SG for this technique is much lower than those of other reported techniques; to the best of our knowledge, this is the first detection approach for PSA. Moreover, these electrocatalysts show promise for the quantification of SG and PSA in aquatic products. Our insights and findings can serve as guidelines for the development of highly active electrocatalysts for application in next-generation food analysis sensors.

Extracellular Vesicles Derived from Bone Mesenchymal Stem Cells Carrying circ_0000075 Relieves Cerebral Ischemic Injury by Competitively Inhibiting miR-218-5p and Up-regulating E3 Ubiquitin Ligase SMURF2.

Extracellular vesicle (EV)-encapsulated circRNAs have the potential role in affecting brain disorders. However, the role of circ_0000075 in cerebral ischemic injury remains unclear. Here, we tried to investigate the mechanism of bone marrow mesenchymal stem cell (BMSC)-derived EVs carrying circ_0000075 in the control of cerebral ischemic injury. Initially, a mouse model with cerebral ischemic injury was induced by middle cerebral artery occlusion (MCAO), followed by the determination of circ_0000075 expression. Then, neurons were isolated and subjected to oxygen-glucose deprivation/reperfusion. BMSCs were isolated for extraction of EVs. The correlation among circ_0000075, microRNA (miR)-218-5p, and Smad ubiquitination regulatory factor 2 (SMURF2) was detected with their roles in cerebral ischemic injury analyzed in vivo and in vitro. circ_0000075 was down-regulated in MCAO mice and engineered RVG-EVs were internalized by neurons to up-regulate circ_0000075 expression. Treatment of RVG-circ_0000075-EVs reduced brain tissue damage, increased neuronal count, and significantly curtailed apoptosis rate, suppressing cerebral ischemic injury in vitro and in vivo. miR-218-5p was targeted by circ_0000075 in neurons, which promoted SMURF2 expression. A negative correlation between SMURF2 and transcriptional regulator Yin Yang 1 (YY1) was identified. In vitro experiments further proved that circ_ 00,000 75 could down-regulate the expression of YY1 through SMURF2, and finally relieving cerebral ischemic injury. Collectively, engineered EVs delivered circ_0000075 into brain tissues and increased circ_0000075 expression, which down-regulated miR-218-5p and up-regulated SMURF2, thus alleviating cerebral ischemic injury.

Development of a highly efficient prime editor 2 system in plants.

Low efficiency has seriously restricted the application of prime editing (PE) systems in plants. In this study, we develop an enhanced plant prime editor 2 system, enpPE2, by stacking various optimization strategies, including updating the PE architecture to PEmax and expressing engineered pegRNA with a structured motif under the control of a composite promoter. In T0 rice plants, enpPE2 exhibits editing frequencies of 64.58% to 77.08%, which are much higher than the frequencies with unmodified pPE2. Our results indicate that the enpPE2 system provides a robust and powerful tool for the precise modification of plant genomes.

Simultaneous enantioselective analysis of illicit drugs in wastewater and surface water by chiral LC-MS/MS: A pilot study on a wastewater treatment plant and its receiving river.

An enantioselective method for quantifying amphetamine-type chiral illicit drugs (CIDs) in wastewater and surface water was developed, validated, and applied to samples from a wastewater treatment plant (WWTP) and its effluent-receiving river in Beijing, China. Water samples were subjected to solid-phase extraction (SPE) and then quantified via liquid chromatography-tandem mass spectrometry. The enantioseparation of CIDs was performed with a CHIRALPAK CBH column. Chromatographic parameters, including mobile phase composition and flow rates, were tested to identify the satisfactory enantiomeric resolution. The SPE method was optimized by evaluating variables, including SPE cartridge types, extraction solvents, and solvent volumes. The Oasis HLB sorbent showed good performance with recoveries exceeding 60% and matrix effects ranging from -19.6% to 26.6% for most target enantiomers, except for norephedrine (NE), in three different aquatic matrixes. The established method was superior to previously reported methods and had a low limit of detection, low limit of quantification, and short runtime (<45 min). The repeatability and reproducibility of the method reached 19.1% and 17.8%, respectively. The method was successfully utilized to monitor the daily variations in CIDs in the influent, effluent, and effluent-receiving river of a WWTP in Beijing over 1 week. The common occurrence of 1 R,2 S-(-)-ephedrine (1 R,2 S-(-)-EPH), 1 S,2 S-(+)-pseudoephedrine (1 S,2 S-(+)-PEPH), R-(-)-methamphetamine (METH), and S-(+)-METH in wastewater samples was observed. Ephedrines (1 R,2 S-(-)-EPH and 1 S,2 S-(+)-PEPH) were the most abundant CIDs in the influent, effluent, upstream, and downstream samples with concentrations of 725.8 ± 181.2 ng/L, 22.9 ± 4.9 ng/L, 12.96 ± 0.79 ng/L, and 11.6 ± 6.7 ng/L, respectively. METH was detectable in most water samples and was present in excess in S-enantiomer form in the influent and in R-enantiomer form in the effluent and surface water. R-(-)-MDMA was detected at a concentration of up to 2.4 ng/L in the influent. The metabolites norketamine (NK), amphetamine(AMP), MDA, and NE were not detected in water samples given the low concentration of their parent drugs.

Erratum: MicroRNA-181 inhibits the proliferation, drug sensitivity and invasion of human glioma cells by targeting Selenoprotein K (SELK).

[This corrects the article on p. 6632 in vol. 11, PMID: 31737213.].

Effects of Phthalate Esters on Ipomoea aquatica Forsk. Seedlings and the Soil Microbial Community Structure under Different Soil Conditions.

Phthalate acid esters (PAEs) are the most frequently utilized synthetic chemical compounds worldwide. They are typical emergent contaminants and are currently attracting considerable concern due to their risks to plants, animals, and public health. Determining the vital environmental factors that affect the toxicity of target pollutants in soil is important for vegetable production and the maintenance and control of soil productivity. We investigated the influence of di-n-butyl phthalate (DBP) and bis(2-ethylhexyl) phthalate (DEHP) under different soil conditions on physiological changes in water spinach (Ipomoea aquatic Forsk.) seedlings and the rhizosphere soil microbial community. Supported by our former experiments in which we determined the representative concentrations that caused the most pronounced toxic effects, three experimental concentrations were studied including control soils without PAEs and spiked soils with either 20 mg DBP or DEHP kg-1 soil. The soil at all the three PAE concentrations was then adjusted to test two soil pH values, three levels of soil organic matter (SOM) content, and three levels of soil moisture content; thus, we completed 12 treatments or conditions simulating different soil environment conditions in greenhouses. After 30 days of cultivation, we analyzed the toxicity effects of two target PAEs on plant growth and physiological factors, and on soil microbial community characteristics. The toxicity of soil DBP and DEHP to the physiology of water spinach was found to be most affected by the soil pH value, then by SOM content, and least of all by soil moisture. The results of the 454 high-throughput sequencing analysis of the soil microbial community indicated that the toxicity of target PAEs to soil microorganisms was most affected by SOM content and then by soil moisture, and no clear relationship was found with soil pH. Under different soil conditions, declines in leaf biomass, chlorophyll a content, and carotenoid content-as well as increases in free amino acid (FAA) content, superoxide anion free radical activity, and hydroxyl radical activity-occurred in response to DBP or DEHP. Heavy use of chemical fertilizer, organic fertilizer, and high humidity led to the special environmental conditions of greenhouse soil, constituting the main conditions considered in this study. The results indicate that under the special highly intensive production systems of greenhouses, soil conditions may directly influence the effects of pollutant phytotoxicity and may thus endanger the yield, nutrient content, and food safety of vegetables. The combined studies of the impacts on plants and rhizosphere microorganisms give a more detailed picture of the toxic effects of the pollutants under different soil conditions.

Phthalate Ester Contamination in Intensively Managed Greenhouse Facilities and the Assessment of Carcinogenic and Non-Carcinogenic Risk: A Regional Study.

The contamination status and the potential carcinogenic and non-carcinogenic health risks from six phthalate esters (PAEs), nominated as priority pollutants by the United States Environmental Protection Agency (USEPA), were investigated in 40 typical greenhouses in three large-scale intensive greenhouse production areas in Jingmen city, Hubei province, central China. The total concentrations of PAEs in 40 soil samples and 80 vegetable samples ranged from 919 ± 134 to 7015 ± 475 µg kg-1 (dry weight, DW), and from 387 ± 63, to 11,683 ± 1313 µg kg-1 (DW), respectively. No carcinogenic risk was detected. The heat-map of the hazard quotient (HQ) values indicates the non-carcinogenic risks to children from di-n-butyl phthalate (DBP), at two sampling sites out of the 40, and from diethylhexyl phthalate (DEHP) (20 to young children and three to older children and adults) at 23 of the sites. The contamination risk from PAEs at Pengdun is of concern because only two of the 14 sampling sites selected there showed the non-carcinogenic risk to humans was unclear. The results of this study help to close a long-term knowledge gap resulting from a shortage of experimental data on PAE contamination in intensive greenhouse vegetable production in central China. The inclusion of DEHP in the Chinese list of priority pollutants is recommended, due to its increasing contamination and risk. This study provides valuable information for protected agricultural soil management and risk avoidance. It is a timely reminder to take PAE contamination and associated health risks into consideration, during the planning and introduction of intensively-managed greenhouse production systems.

MicroRNA-181 inhibits the proliferation, drug sensitivity and invasion of human glioma cells by targeting Selenoprotein K (SELK).

Gliomas are aggressive type of brain tumors and cause significant human mortality world over. The frequent relapses, development of drug resistance, the adverse effects of the chemotherapy and dearth of the therapeutic targets form the major hurdles in glioma treatment. Several studies suggest that microRNAs (miRs) are involved in the development and progression of different cancers. Herein, the therapeutic potential of miR-181 was explored in human glioma cells. The results showed that miR-181 is significantly downregulated in human glioma cells. Overexpression of miR-181 caused significant inhibition in the proliferation of U87 and U118 glioma cells. The miR-181 triggered growth inhibition was found to be mainly due to the induction of apoptosis which was concomitant with increase in the Bax/Bcl-2 ratio. Additionally, miR-181 enhanced the chemosensitivity of the glioma cells to temozolomide and suppressed their invasion. Bioinformatic analysis showed that miR-181 exerts its effects by inhibiting the expression of Selenoprotein K (SELK). The expression of SELK was found to be significantly upregulated in glioma cells and silencing of SELK suppressed the proliferation of glioma cells. Nonetheless, overexpression of SELK could nullify the effects of miR-181 on the proliferation of the glioma cells. Taken together, miR-181 may exhibit therapeutic implications in the treatment of glioma.

[Effect of various intervention factors on MMP-3 and TIMP-1 level in synovial fluid in knee joints with osteroarthritis].

OBJECTIVE: To examine the expression of MMP-3 and TIMP-1 in the synovial fluid in knee joints with osteoarthritis before and after being treated with hyaluronic acid(HA), glucosamine sulfate(GS) and arthroscopic de bridment(AD), and to explore the therapeutic mechanism. METHODS: Sixty patients (64 knees) with osteoarthritis(OA) were randomly divided into HA group AD group and GS+AD group. Some patients from the HA group and the GS+HA group were selected, and served as HA' group and GS+HA' group. The expression of MMP-3 and TIMP-1 in the synovial fluid was measured by enzyme-linked immunosorbent assay (ELISA) before and after 4 week and 6 month therapy. RESULTS: The level of MMP-3 and the ratio of MMP-3/TIMP-1 in the synovial fluid decreased after being treated for 4 weeks, and the effect on MMP-3 and MMP-3/TIMP-1 lasted for 6 months in the HA group and the GS+HA groups. The levels of TIMP-1 increased significantly after being treated for 4 weeks only in the GS+HA group. The level of MMP-3 and the ratio of MMP-3/TIMP-1 in the synovial fluid decreased, but the level of TIMP-1 increased after being treated for 4 weeks, and the effect on MMP-3 and MMP-3/TIMP-1 lasted for 6 months. The level of MMP-3 and the ratio of MMP-3/TIMP-1 in the synovial fluid increased after being treated for 6 months compared with those for 4 weeks. The level of TIMP-1 increased in the GS group more than that in the HA group after being treated for 4 weeks. The level of TIMP-1 increased in the AD group more than that in the HA' group and the GS+HA' group after being treated for 4 weeks. CONCLUSION: (1) HA, GS and AD all can decrease the level of of MMP-3 and the ratio of MMP-3/TIMP-1 in the synovial fluid in knee joints with OA. (2) The level of TIMP-1 in the synovial fluid has no difference before and after being treated with HA. The AD group and GS group can increase the level of TIMP-1, which indicates that the AD group or GS group might produce better therapeutic effect. (3) The level of TIMP-1 increased in the AD group is more than that in the HA' group and the GS+HA' group after being treated for 4 weeks, which indicates that the AD group might get better therapeutic effect than the other 2 groups. (4) One of the most important mechanisms of HA, GS and AD in treating OA might be attributed to the expression of MMPs and TIMPs in knee joints with OA.

Application of catastrophe theory in comprehensive ecological security assessment of plastic greenhouse soil contaminated by phthalate esters.

Large amount of phthalate esters (PAEs) used as plasticizers in polyvinyl chloride (PVC) products has caused ubiquitous contamination to the environment and potential ecology security risk all around the world, especially in places plastic films were indispensably utilized due to the widely proposing of facility agriculture in China. A case of PAEs contamination in four suburb areas of Nanjing was analyzed and discussed in this study. A new frame work has been put forward based on multi-criteria evaluation model and mathematical method of catastrophe theory, using farming work, laboratory determination and relevant environmental standards to measure the ecology security risk of PAEs in study areas. The factors were selected based on the availability of the data and the local conditions. The assessment model involves the contamination status of PAEs in soil and vegetables, the contamination effects of PAEs to human and soil organisms and the contamination source of PAEs from plastic films and other products in the four study facility agriculture areas. An evaluation system of the model was composed of thirteen mesosphere indicators and twenty-five underlying indicators including total PAEs concentration in soils, single PAE concentration in soils, total PAEs concentrations in roots, leafy, solanaceous and stem vegetables, PAE human risks, soil microbial counts, microorganism diversity indices, atmospheric deposition of PAEs, whether sewage wastewater irrigation, planting mode of the facility agriculture areas and climate condition of study areas. The modified evaluation system was used in the assessment of ecology security of the same place based on the data of 2012, and the results suggested that the ecology security indicators were reliable and were agree well with the practical situation of the study areas. The results could provide guidance for the application of health risk assessment of soil environment for the strong objectivity of catastrophe theory compared with other evaluation methods.