The activities and mechanisms of intestinal microbiota metabolites of TCM herbal ingredients could be illustrated by a strategy integrating spectrum-effects, network pharmacology, metabolomics and molecular docking analysis: Platycodin D as an example.

BACKGROUND: The intestinal microbiota plays a key role in understanding the mechanism of traditional Chinese medicine (TCM), as it could transform the herbal ingredients to metabolites with higher bioavailability and activity comparing to their prototypes. Nevertheless, the study of the activity and mechanism of microbiota metabolites reported by the published literature still lacks viable ways. Hence a new strategy is proposed to solve this issue. PURPOSE: A new strategy to study the activity and mechanism of intestinal microbiota metabolites of TCM herbal ingredients by integrating spectrum-effect relationship, network pharmacology, metabolomics analysis and molecular docking together was developed and proposed. METHOD: Platycodin D (PD) and its microbiota metabolites with antitussive and expectorant effect were selected as an example for demonstration. First, the PD and its microbiota metabolites with important contribution to antitussive and/or expectorant effects were screened through spectrum-effect relationship analysis. Second, network pharmacology and metabolomics analysis were integrated to identify the upstream key targets of PD and its microbiota metabolites as well as the downstream endogenous metabolites. Finally, the active forms of PD were further confirmed by molecular docking. RESULTS: Results showed that PD was an active ingredient with antitussive and/or expectorant effects, and the active forms of PD were its microbiota metabolites: 3-O-ß-d-glucopyranosyl platycodigenin, 3-O-ß-d-glucopyranosyl isoplatycodigenin, 7­hydroxyl-3-O-ß-d-glucopyranosyl platycodigenin, platycodigenin and isoplatycodigenin. In addition, those microbiota metabolites could bind the key targets of PAH, PLA2G2A, ALOX5, CYP2C9 and CYP2D6 to exert antitussive effects by regulating four metabolic pathways of phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism, glycerophospholipid metabolism and linoleic acid metabolism. Similarly, they could also bind the key targets of PLA2G1B, ALOX5, CYP2C9 and CYP2D6 to exert expectorant effect by regulating two pathways of glycerophospholipid metabolism and linoleic acid metabolism. CONCLUSION: The proposed strategy paves a new way for the illustration of the activities and mechanisms of TCM herbal ingredients, which is very important to reconcile the conundrums of TCM herbal ingredients with low oral bioavailability but high activity.

LOX-1 Regulation in Anti-atherosclerosis of Active Compounds of Herbal Medicine: Current Knowledge and the New Insight.

Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) have recently been identified to be closely related to the occurrence and development of atherosclerosis (AS). A growing body of evidence has suggested Chinese medicine takes unique advantages in preventing and treating AS. In this review, the related research progress of AS and LOX-1 has been summarized. And the anti-AS effects of 10 active components of herbal medicine through LOX-1 regulation have been further reviewed. As a potential biomarker and target for intervention in AS, LOX-1 targeted therapy might provide a promising and novel approach to atherosclerotic prevention and treatment.

FveARF2 negatively regulates fruit ripening and quality in strawberry.

Auxin response factors (ARFs) are transcription factors that play important roles in plants. ARF2 is a member of the ARF family and participates in many plant growth and developmental processes. However, the role of ARF2 in strawberry fruit quality remains unclear. In this study, FveARF2 was isolated from the woodland strawberry 'Ruegen' using reverse transcription-polymerase chain reaction (RT-PCR), which showed that FveARF2 expression levels were higher in the stem than in other organs of the 'Ruegen' strawberry. Moreover, FaARF2 was higher in the white fruit stage of cultivated strawberry fruit than in other stage. Subcellular localization analysis showed that FveARF2 is located in the nucleus, while transcriptional activation assays showed that FveARF2 inhibited transcription in yeast. Silencing FveARF2 in cultivated strawberry fruit revealed earlier coloration and higher soluble solid, sugar, and anthocyanin content in the transgenic fruit than in the control fruit, overexpression of FveARF2 in strawberry fruit delayed ripening and lower soluble solid, sugar, and anthocyanin content compared to the control fruit. Gene expression analysis indicated that the transcription levels of the fruit ripening genes FaSUT1, FaOMT, and FaCHS increased in FveARF2-RNAi fruit and decreased in FveARF2-OE fruit, when compared with the control. Furthermore, yeast one-hybrid (Y1H) and GUS activity experiments showed that FveARF2 can directly bind to the AuxRE (TGTCTC) element in the FaSUT1, FaOMT, and FaCHS promoters in vitro and in vivo. Potassium ion supplementation improved the quality of strawberry fruit, while silencing FveARF2 increased potassium ion content in transgenic fruit. The Y1H and GUS activity experiments also confirmed that FveARF2 could directly bind to the promoter of FveKT12, a potassium transporter gene, and inhibited its expression. Taken together, we found that FveARF2 can negatively regulate strawberry fruit ripening and quality, which provides new insight for further study of the molecular mechanism of strawberry fruit ripening.

[Suitable harvest period of Aurantii Fructus based on UPLC-Q-TOF-MS metabolomics].

The types of secondary metabolites of Aurantii Fructus samples from GAP base in different harvest periods were detected by UPLC-Q-TOF-MS metabolomics, and the differential metabolites were screened out by multivariate statistical analysis. The variation of the content of differential metabolites with different harvest periods was analyzed, and the correlation analysis was carried out on the differential metabolites to determine the suitable harvest period for different components. Sixteen differential metabolites were obtained. With the delay of harvest time, the content of flavonoid glycosides, including naringin, neohesperidin, poncirin, narirutin, and hesperidin, gradually decreased. It is suggested that the suitable harvest period for raw materials of Aurantii Fructus with flavonoids as active components is from July 18 to July 25(within one week before and after the Great heat). The content of nobiletin, tangeretin, natsudaidain, 7-hydroxyl-4',3,5,6,8-pentamethoxyflavone, sinensetin, isosinensetin, 5,6,7,4'-tetramethoxyflavone, and isomeranzin decreased first, then increased, and finally decreased. It is suggested that the suitable harvest time for raw materials of Aurantii Fructus with these components as the active components is July 18. The content changes of meranzin, limonin, and 3,5,6,7,8,3',4'-heptamethoxyflavone have their characteristics. According to the conditions of actual production, it is suggested that the suitable harvest time is June 27, July 11, and July 25, respectively. The results showed that there were differences in the content of chemical components of Aurantii Fructus in different harvest periods, and the suitable harvest period should be determined according to the differences in chemical component content. This study is expected to provide a scientific basis for the purchase of raw materials of Aurantii Fructus for Chinese patent medicines with different effects.

Cerebralcare Granule® combined with nimodipine improves cognitive impairment in bilateral carotid artery occlusion rats by reducing lipocalin-2.

AIMS: Clinically, Cerebralcare Granule® (CG) has been widely utilized to treat various types of headache, chronic cerebral insufficiency and other diseases, and the effect is significant. Clinical studies have shown that CG can significantly relieve vascular dementia (VaD), however, the molecular mechanisms haven't been established. To clear the therapeutic mechanisms of CG against VaD, a hypothesis was proposed that CG could treat neurovascular injury by inhibiting the production of lipocalin-2 (LCN 2). MAIN METHODS: 90 dementia rats were selected by water maze test and randomly divided into 6 groups, including nimodipine (NM), CG L (low dose) (0.314 g kg-1), CG H (high dose) (0.628 g kg-1), and combined group (CG + NM). And in vitro neuronal cell OGD modeling to evaluate the effect of CG on JAK2/STAT3. KEY FINDINGS: CG could significantly shorten the escape latency of two-vessel occlusion (2-VO) rats, increase their exploratory behavior, alleviate the symptoms of VaD and improve the ultrastructural pathological damage of neurovascular unit and accelerate the recovery of cerebral blood perfusion. CG combined with NM is better than NM alone. It was further showed that CG could inhibit the pathogenicity of LCN 2 through JAK2/STAT3 pathway and suppress the production of inflammatory cytokines. It plays a role in the protection of cerebral microvasculature and BBB in 2-VO rats. SIGNIFICANCE: Taken together, there data has supported notion that CG can protect the integrity of cerebral blood vessels and BBB and improve cognitive impairment through mainly inhibiting LCN 2, which provides scientific evidence for clinical application.

A Novel PL9 Pectate Lyase from Paenibacillus polymyxa KF-1: Cloning, Expression, and Its Application in Pectin Degradation.

Pectate lyases play an important role in pectin degradation, and therefore are highly useful in the food and textile industries. Here, we report on the cloning of an alkaline pectate lyase gene (pppel9a) from Paenibacillus polymyxa KF-1. The full-length gene (1350 bp) encodes for a 449-residue protein that belongs to the polysaccharide lyase family 9 (PL9). Recombinant PpPel9a produced in Escherichia coli was purified to electrophoretic homogeneity in a single step using Ni2+-NTA affinity chromatography. The enzyme activity of PpPel9a (apparent molecular weight of 45.3 kDa) was found to be optimal at pH 10.0 and 40 °C, with substrate preference for homogalacturonan type (HG) pectins vis-à-vis rhamnogalacturonan-I (RG-I) type pectins. Using HG-type pectins as substrate, PpPel9a showed greater activity with de-esterified HGs. In addition, PpPel9a was active against water-soluble pectins isolated from different plants. Using this lyase, we degraded citrus pectin, purified fractions using Diethylaminoethyl (DEAE)-sepharose column chromatography, and characterized the main fraction MCP-0.3. High-performance gel permeation chromatography (HPGPC) analysis showed that the molecular mass of citrus pectin (~230.2 kDa) was reduced to ~24 kDa upon degradation. Ultra-performance liquid chromatography - tandem mass spectrometer (UPLC-MS) and monosaccharide composition analyses demonstrated that PpPel9a worked as an endo-pectate lyase, which acted primarily on the HG domain of citrus pectin. In vitro testing showed that the degradation product MCP-0.3 significantly promotes the growth of Lactobacillus plantarum and L. rhamnosus. In this regard, the enzyme has potential in the preparation of pharmacologically active pectin products.

[Effects of nitrogen additions on soil hydrolase and oxidase activities in Pinus elliottii plantations.] / æ°®æ·»åŠ å¯¹æ¹¿åœ°æ¾æž—åœŸå£¤æ°´è§£é ¶å’Œæ°§åŒ–é ¶æ´»æ€§çš„å½±å“.

We evaluated responses of hydrolase and oxidase activities in a subtropical Pinus elliottii plantation through a nitrogen (N) addition field experiment (dosage level: 0, 40, 120 kg N·hm-2·a-1). The results showed that N additions significantly decreased the carbon, nitrogen and phosphorus related hydrolase and oxidase activities. The activities of ß-1,4-glucosidase (BG), cellobiohydrolase (CBH), ß-1,4-N-acetylglucosaminidase (NAG) and peroxidase (PER) activities were decreased by 16.5%-51.1% due to N additions, and the decrease was more remarkable in the higher N addition treatment. The activities of α-1,4-glucosidase (aG), ß-1,4-xylosidase (BX), acid phosphatase (AP) and phenol oxidase (PPO) were decreased by 14.5%-38.6% by N additions, however, there was no significant difference among the different N addition treatments. Soil enzyme activities varied obviously in different seasons. The activities of BG, NAG, BX, CBH, AP and PPO were in the order of March > June > October, and aG and PER activities were in the order of October > March > June. Most of the soil hydrolase and oxidase activities were positively correlated with soil pH, but negatively with NO3--N content. It indicated that N additions inhibited soil hydrolase and oxidase activities by reducing soil pH and increasing soil nitrification. N additions inhibited the soil organic matter mineralization and turnover in the subtropical area, and the effects were obvious with the increasing dosage of N additions.

Berberine Attenuates Vascular Remodeling and Inflammation in a Rat Model of Metabolic Syndrome.

Berberine is a natural product that shows benefits for metabolic syndrome (MS). However, the effects of berberine on the improvement of vascular inflammation and remodeling in MS remain unclear. This study aimed to investigate whether berberine could prevent vascular remodeling and inflammation in the MS condition. A rat model of MS was established, and MS rats were divided into two groups: MS group without berberine treatment, and MSB group with berberine treatment (each group n-10). Ten normal Wistar rats were used as controls (NC group). Vascular damage was examined by transmission electron microscopy and pathological staining. Compared to the NC group, the secretion of inflammatory factors was increased and the aortic wall thicker in the MS group. The MSB group exhibited decreased secretion of inflammatory factors and improved vascular remodeling, compared to the MS group. In addition, the levels of p38 mitogen-activated protein kinase (p38 MAPK), activating transcription factor 2 (ATF-2) and matrix metalloproteinase 2 (MMP-2) were significantly decreased in the MSB group compared to the MS group. In conclusion, our data show that berberine improves vascular inflammation and remodeling in the MS condition, and this is correlated with the ability of berberine to inhibit p38 MAPK activation, ATF-2 phosphorylation, and MMP-2 expression.

[Effects of different fertilizer species on carbon and nitrogen leaching in a reddish paddy soil].

Enhanced fertilization could decrease nitrogen utilization rate and increase carbon and nitrogen leaching, leading to water pollution in agricultural ecosystem. A long-term field experiment had been established on a reddish paddy soil of Qianyanzhou Ecological Experimental Station (114 degrees 53'E, 26 degrees 48'N) in Jiangxi Province in 1998. Soil solution samples were collected by clay tube and vacuum pump. Four fertilizer species treatments were selected: control with no fertilizer (CK), straw return (ST), nitrogen, phosphorus and potassium mineral fertilizers (NPK) and pig manure (OM), aiming to evaluate the effects of different species of fertilizer on carbon and nitrogen leaching in a double rice cropping system. The results showed that: (1) ammonium nitrogen (NH4(+) -N) was the major type of N in soil leachate in reddish paddy soil. The application of NPK could significantly increase the ammonium nitrogen concentration (1.2 mg x L(-1) +/- 0.1 mg x L(-1)) compared with the CK, ST and OM treatments, and the application of OM could significantly increase the dissolved organic carbon (DOC) concentration (27.3 mg x L(-1) +/- 1.6 mg x L(-1)) in soil leachate. The carbon and nitrogen leaching were more notable in the vegetative growth stage than the reproductive growth stage of rice (P < 0.05); (2) the long-term application of NPK and OM increased the NH4(+) -N, DOC, soil organic carbon (SOC) and total nitrogen (TN) contents. The NPK was best beneficial to improve TN contents and OM to improve SOC contents. (3) The DOC contents in soil leachate and SOC in paddy soil had a positive correlation (P < 0.01), while NH4(+) -N contents in soil leachate and TN contents in paddy soil had a positive correlation (P < 0.01).

Efficacy and safety of sorafenib for advanced non-small cell lung cancer: a meta-analysis of randomized controlled trials.

BACKGROUND: Many clinical trials have been conducted to evaluate sorafenib for the treatment of advanced NSCLC, but the results for efficacy have been inconsistent. The aim of this study was to evaluate the efficacy and safety of sorafenib in patients with advanced NSCLC in more detail by meta-analysis. METHODS: This meta-analysis of randomized controlled trials (RCTs) was performed after searching PubMed, EMBASE, ASCO Abstracts, ESMO Abstracts, and the proceedings of major conferences for relevant clinical trials. Two reviewers independently assessed the quality of the trials. Outcomes analysis were disease control rate (DCR), progression- free survival (PFS), overall survival (OS) with 95% confidence intervals (CI) and major toxicity. Subgroup analysis was conducted according to sorafenib monotherapy, in combination with chemotherapy or EGFR-TKI to investigate the preferred therapy strategy. RESULTS: Results reported from 6 RCTs involving 2,748 patients were included in the analysis. Compared to sorafenib-free group, SBT was not associated with higher DCR (RR 1.31 (0.96-1.79), p=0.09), PFS (HR 0.82 (0.66-1.02), p=0.07) and OS (HR 1.01 (0.92-1.12), p=0.77). In terms of subgroup results, sorafenib monotherapy was associated with significant superior DCR and longer PFS, but failed to show advantage with regard to OS. Grade 3 or greater sorafenib-related adverse events included fatigue, hypertension, diarrhea, oral mucositis, rash and HFSR. CONCLUSIONS: SBT was revealed to yield no improvement in DCR, PFS and OS. However, sorafenib as monotherapy showed some activity in NSCLC. Further evaluation may be considered in subsets of patients who may benefit from this treatment. Sorafenib combined inhibition therapy should be limited unless the choice of platinum-doublet regimen, administration sequence or identification of predictive biomarkers are considered to receive better anti-tumor activity and prevention of resistance mechanisms.

[Effects of applying different kind fertilizers on enzyme activities related to carbon, nitrogen, and phosphorus cycles in reddish paddy soil].

Based on the long-term fixed position experimental data from Qianyanzhou Ecological Experiment Station, Chinese Academy of Sciences in 1998, this paper analyzed the effects of applying different kind fertilizers (straw, ST; pig manure, OM; and chemical fertilizer, NPK) on the nutrients (C, N, and P) status and the activities of related enzymes ( beta-1,4-glucosidase, betaG; beta-1,4-N-acetylglucosaminidase, NAG; L-leucine aminopeptidase, LAP; and acid phosphatase, AP) in reddish paddy soil. With the application of OM, the activities of soil betaG, NAG, and LAP increased significantly, as compared with other treatments, and were 1.4, 2. 6, and 1.9 times higher than the control (CK) , respectively. Applying OM also improved the ratio of soil organic carbon to total nitrogen (C/N), but decreased the soil betaG/(NAG+LAP) ratio, suggesting that pig manure could benefit the degradation of soil cellulose and the accumulation of soil organic carbon. Applying NPK increased the activities of soil betaG, NAG, and LAP, but decreased the AP activity, with a decrement of 34% as compared with CK. Under the application of NPK, the soilbetaG/AP and (NAG+ LAP)/AP ratios increased, but the ratios of soil organic carbon to total phosphorus (C/P) and of soil total nitrogen to total phosphorus (N/P) decreased, indicating that chemical fertilizers could induce the accumulation of soil inorganic phosphorus, and inhibit the microbial functions of degrading polysaccharides and phosphate phospholipids.

ICPS: an integrative cancer profiler system.

UNLABELLED: Founded upon the database of 570 public signatures, ICPS is a web-based application to obtain biomarker profiles among 11 common cancers by integrating genomic alterations with transcription signatures on the basis of a previously developed integrative pipeline. ICPS supports both public data and user's in-house data, and performs meta-analysis at a cancer subtype level by combining heterogeneous datasets. Finally, ICPS returns the robust gene signature containing potential cancer biomarkers that may be useful to carcinogenesis study and clinical cancer diagnosis. AVAILABILITY: http://server.bioicps.org CONTACT: zhxy@mail.tsinghua.edu.cn; zxy-dcs@mail.tsinghua.edu.cn

Allele-specific inhibition of divergent protein tyrosine phosphatases with a single small molecule.

A central challenge of chemical biology is the development of small-molecule tools for controlling protein activity in a target-specific manner. Such tools are particularly useful if they can be systematically applied to the members of large protein families. Here we report that protein tyrosine phosphatases can be systematically 'sensitized' to target-specific inhibition by a cell-permeable small molecule, Fluorescein Arsenical Hairpin Binder (FlAsH), which does not inhibit any wild-type PTP investigated to date. We show that insertion of a FlAsH-binding peptide at a conserved position in the PTP catalytic-domain's WPD loop confers novel FlAsH sensitivity upon divergent PTPs. The position of the sensitizing insertion is readily identifiable from primary-sequence alignments, and we have generated FlAsH-sensitive mutants for seven different classical PTPs from six distinct subfamilies of receptor and non-receptor PTPs, including one phosphatase (PTP-PEST) whose three-dimensional catalytic-domain structure is not known. In all cases, FlAsH-mediated PTP inhibition was target specific and potent, with inhibition constants for the seven sensitized PTPs ranging from 17 to 370 nM. Our results suggest that a substantial fraction of the PTP superfamily will be likewise sensitizable to allele-specific inhibition; FlAsH-based PTP targeting thus potentially provides a rapid, general means for selectively targeting PTP activity in cell-culture- or model-organism-based signaling studies.

[Identification of critical area of phosphorus loss in agricultural areas of Guishui River watershed by phosphorus loss risk assessment].

Agricultural non-point sources pollution is one of severe problems for water environment of agricultural areas in China. Because of the big difficulties, identifying the critical source areas for phosphorus loss becomes the focal point of the non-point sources pollution control. A modified catchment scale phosphorus ranking scheme was developed for agricultural areas in Guishui River watershed. The new scheme included eight assessment factors, which had three phosphorus loss risk ranks respectively and selected quantitative analysis method. The result shows that the phosphorus fertilizer management of the vegetable fields is the most unfit method and has high phosphorus loss probabilities. Most study areas have high soil available phosphorus content and low soil erosion degree. The figure of the assessment result shows that the areas that are categorized as "low" phosphorus loss risk are small. Based on the figure of the result, the critical source areas were confirmed and the management strategies were brought forward according to the analysis on the distribute characteristics of the critical source areas. The result shows that the modified catchment scale phosphorus ranking scheme has operability and practicability to a certain extent.

[Catchment scale risk assessment and critical source area identification of agricultural phosphorus loss].

Agricultural non-point source phosphorus pollution is a severe problem for rural water bodies in China, but hard to control directly because of its special characteristics. In this paper, an approach on the catchment scale risk assessment and critical source area identification of agricultural phosphorus loss in northern China was made, based on the catchment scale phosphorus ranking scheme and the method proposed by Gburek et al. Eight factors were selected and weighed in the modified catchment scale phosphorus ranking scheme, and the phosphorus loss risk rating of each factor was adjusted based on the current professional standards and the actual circumstances in China. The areas with ' high' risk rating of phosphorus loss in definite catchment were the critical source areas for non-point source phosphorous pollution control in that catment. The availability of obtained data and the quantification of the assessment were taken into account in the new scheme, and GIS technique and geostatistics were used for confirming the factors. Therefore, the new scheme had definite operability and practicability.

Assessing field vulnerability to phosphorus loss in Beijing agricultural area using Revised Field Phosphorus Ranking Scheme.

Guanting Reservoir, one of the drinking water supply sources of Beijing, suffers from water eutrophication. It is mainly supplied by Guishui River. Thus, to investigate the reasons of phosphorus (P) loss and improve the P management strategies in Guishui River watershed are important for the safety of drinking water in this region. In this study, a Revised Field P Ranking Scheme (PRS) was developed to reflect the field vulnerability of P loss at the field scale based on the Field PRS. In this new scheme, six factors are included, and each one was assigned a relative weight and a determination method. The affecting factors were classified into transport factors and source factors, and, the standards of environmental quality on surface water and soil erosion classification and degradation of the China were used in this scheme. By the new scheme, thirty-four fields in the Guishui River were categorized as "low", "medium" or "high" potential for P loss into the runoff. The results showed that the P loss risks of orchard and vegetable fields were higher than that of corn and soybean fields. The source factors were the main factors to affect P loss from the study area. In the study area, controlling P input and improving P usage efficiency are critical to decrease P loss. Based on the results, it was suggested that more attention should be paid on the fields of vegetable and orchard since they have extremely high usage rate of P and high soil test of P. Compared with P surplus by field measurements, the Revised Field PRS was more suitable for reflecting the characteristics of fields, and had higher potential capacity to identify critical source areas of P loss than PRS.

[Enrichment of denitrifying phosphate accumulating organisms in sequencing batch membrane bioreactors].

The enrichment and characteristics of denitrifying phosphate accumulating organisms (DPAO), which are capable of utilizing nitrate as electron acceptor, was investigated in a laboratory-scale sequencing batch membrane bioreactors (SBMBR). The results demonstrated that the proportion of DPAO increased from 19.4% to 69.6% of total phosphate accumulating organisms after anaerobic-aerobic and anaerobic-anoxic-aerobic phases. SBMBR system could operate steadily when 120 mg nitrate was added continuously during the anoxic phase every period. Simultaneous phosphate uptake and biological denitrification with good performance could be obtained in SBMBR operated in steady-state. Nitrate and phosphorus removal efficiency were above 100% and 84% respectively during anoxic phase, however, the effluent TP concentration was low than 0.5 mg/L, the total phosphorus removal efficiency was 96.1%. Furthermore, the ammonium nitrogen removal efficiency of SBMBR could be maintained at 92.2%, and the cumulation of nitrite and nitrate was not observed clearly.

[Effects of land use and management practice on farmland soil quality in Yanhuai basin of Beijing].

In April 2005, a total of 41 soil samples were collected from the sandy loam soil (Ustochrepts) under orchard, vegetable, corn (Zea maize L. ) and soybean (Glycine max L. ) in Yanhuai basin of Beijing, and their physical, chemical, and biological properties were analyzed. The results showed that the contents of soil organic carbon (SOC), total nitrogen (TN) and total phosphorus (TP) in vegetable field, orchard field, and high-input corn field were obviously higher than those in soybean field and mid- and low-input corn fields. In these fields, SOC content varied from 7. 67 to 10. 00 g x kg(-1), TN content varied from 0. 75 to 1. 12 g x kg(-1) , and TP content varied from 0. 63 to 1. 00 g x kg(-1). Soil available phosphorus (AP) content and electric conductance (EC) were significantly higher in vegetable field than in other fields, while no significant differences in soil physical properties were observed among the four land use types. The soil quality index of orchard and vegetable fields was 0. 525 and 0. 503, respectively, being higher than those of corn and soybean fields (0. 417-0. 494). Vegetable field had the highest efficiency of soil nutrients, but non-point source pollution risk should be minded. For soybean field and mid- and low-input corn fields, intensive management should be practiced to improve their soil quality.

Intramammary expression and therapeutic effect of a human lysozyme-expressing vector for treating bovine mastitis.

To develop a gene therapy strategy for treating bovine mastitis, a new mammary-specific vector containing human lysozyme (hLYZ) cDNA and kanamycin resistance gene was constructed for intramammary expression and clinical studies. After one time acupuncture or intracisternal infusion of healthy cows with 400 microg of the p215C3LYZ vector, over 2.0 microg/ml of rhLYZ could be detected by enzymatic assay for about 3 weeks in the milk samples. Western blotting showed that rhLYZ secreted into milk samples from the vector-injected cows had molecular weight similar to that of the natural hLYZ in human colostrums. Twenty days after the primary injection, the quarters were re-injected with the same vector by quarter acupuncture and even higher concentrations of rhLYZ could be detected. Indirect competitive ELISA of milk samples showed that the vector injection did not induce detectable humoral immune response against hLYZ. Clinical studies showed that twice acupuncture of quarters with the p215C3LYZ vector had overt therapeutic effect on clinical and subclinical mastitis previously treated with antibiotics, including disappearance of clinical symptoms and relatively high microbiological cure rates. These data provide a solid rationale for using the vector to develop gene therapy for treating bovine mastitis.