Enhancement growth, water use efficiency and economic benefit for maize by drip irrigation in Northwest China.

The application of drip irrigation has been paid more and more attention, but there was lack of systematic comparative analysis between drip irrigation and conventional border irrigation method for maize, currently. A 7-year field study from 2015 to 2021 evaluated the effects of drip irrigation (DI, 540 mm) or conventional border irrigation method (BI, 720 mm) on maize growth, water use efficiency (WUE) as well as profitability. The results showed the plant height, leaf area index, yield, WUE and economic benefit of maize with DI had significantly higher than BI. The dry matter translocation, the dry matter transfer efficiency and contribution of dry matter translocation to grain with DI showed significant increase of 27.44%, 13.97% and 7.85% compared to BI, respectively. In comparison to conventional border irrigation, the yield of drip irrigation increased by 14.39%, as well as WUE and irrigation water use efficiency (IWUE) increased by 53.77% and 57.89%. The net return and economic benefit of drip irrigation was 1998.87 and 756.58 USD$ hm-1 higher than that of BI. Drip irrigation increased net return and benefit/cost ratio by 60.90% and 22.88% compared with BI. These results demonstrate that the drip irrigation can effectively improve the growth, yield, WUE and economic benefit of maize in northwest China. Therefore, drip irrigation can be used for maize cultivation to increase crop yield and WUE in northwest China, which has cut down on irrigation water about 180 mm.

Multidimensional spatial monitoring of open pit mine dust dispersion by unmanned aerial vehicle.

Dust pollution is one of the most severe environmental issues in open pit mines, hindering green mining development. Open pit mine dust has characteristics of multiple dust-generating points, is irregular, influenced by climatic conditions, and has a high degree of distribution with a wide dispersion range in three dimensions. Consequently, evaluating the quantity of dust dispersion and controlling environmental pollution are crucial for supporting green mining. In this paper, dust monitoring above the open pit mine was carried out with an unmanned aerial vehicle (UAV) on board. The dust distribution patterns above the open pit mine were studied in different vertical and horizontal directions at different heights. The results show that the temperature changes less in the morning and more at noon in winter. At the same time, the isothermal layer becomes thinner and thinner as the temperature rises, which makes it easy for dust to spread. The horizontal dust is mainly concentrated at 1300 and 1550 elevations. The dust concentration is polarized at 1350-1450 elevation. The most serious exceedance is at 1400 elevation, with TSP (the concentration of total suspended particulate), PM10 (particulates with aerodynamic diameter < 10 µm), and PM2.5 (particulates with aerodynamic diameter < 2.5 µm) accounting for 188.8%, 139.5%, and 113.8%, respectively. The height is 1350-1450 elevation. Dust monitoring technology carried out by UAV can be applied to the study of dust distribution in the mining field, and the research results can provide reference for other open pit mines. It can also provide a basis for law enforcement part to carry out law enforcement, which has expanded and wide practical application value.

The Synergistic Effect of Topographic Factors and Vegetation Indices on the Underground Coal Mine Utilizing Unmanned Aerial Vehicle Remote Sensing.

Understanding the synergistic effect between topography and vegetation in the underground coal mine is of great significance for the ecological restoration and sustainable development of mining areas. This paper took advantage of unmanned aerial vehicle (UAV) remote sensing to obtain high-precision topographic factors (i.e., digital elevation model (DEM), slope, and aspect) in the Shangwan Coal Mine. Then, a normalized difference vegetation index (NDVI) was calculated utilizing Landsat images from 2017 to 2021, and the NDVI with the same spatial resolution as the slope and aspect was acquired by down-sampling. Finally, the synergistic effect of topography and vegetation in the underground mining area was revealed by dividing the topography obtained using high-precision data into 21 types. The results show that: (1) the vegetation cover was dominated by "slightly low-VC", "medium-VC", and "slightly high-VC" in the study area, and there was a positive correlation between the slope and NDVI when the slope was more than 5°. (2) When the slope was slight, the aspect had less influence on the vegetation growth. When the slope was larger, the influence of the aspect increased in the study area. (3) "Rapidly steep-semi-sunny slope" was the most suitable combination for the vegetation growth in the study area. This paper revealed the relationship between the topography and vegetation. In addition, it provided a scientific and effective foundation for decision-making of ecological restoration in the underground coal mine.

Metagenomic analysis of the soil microbial composition and salt tolerance mechanism in Yuncheng Salt Lake, Shanxi Province.

The soil in Yuncheng Salt Lake has serious salinization and the biogeographic environment affects the composition and distribution of special halophilic and salt-tolerant microbial communities in this area. Therefore, this study collected soils at distances of 15, 30, and 45 m from the Salt Lake and used non-saline soil (60 m) as a control to explore the microbial composition and salt tolerance mechanisms using metagenomics technology. The results showed that the dominant species and abundance of salt-tolerant microorganisms changed gradually with distance from Salt Lake. The salt-tolerant microorganisms can increase the expression of the Na+/H+ antiporter by upregulating the Na+/H+ antiporter subunit mnhA-G to respond to salt stress, simultaneously upregulating the genes in the betaine/proline transport system to promote the conversion of choline into betaine, while also upregulating the trehalose/maltose transport system encode genes to promote the synthesis of trehalose to resist a high salt environment.

Monitoring of Vegetation Disturbance and Restoration at the Dumping Sites of the Baorixile Open-Pit Mine Based on the LandTrendr Algorithm.

Overstocked dumping sites associated with open-pit coal mining occupy original vegetation areas and cause damage to the environment. The monitoring of vegetation disturbance and restoration at dumping sites is important for the accurate planning of ecological restoration in mining areas. This paper aimed to monitor and assess vegetation disturbance and restoration in the dumping sites of the Baorixile open-pit mine using the LandTrendr algorithm and remote sensing images. Firstly, based on the temporal datasets of Landsat from 1990 to 2021, the boundaries of the dumping sites in the Baorixile open-pit mine in Hulunbuir city were extracted. Secondly, the LandTrendr algorithm was used to identify the initial time and duration of vegetation disturbance and restoration, while the Normalized Difference Vegetation Index (NDVI) was used as the input parameter for the LandTrendr algorithm. Thirdly, the vegetation restoration effect at the dumping sites was monitored and analyzed from both temporal and spatial perspectives. The results showed that the dumping sites of the Baorixile open-pit mine were disturbed sharply by the mining activities. The North dumping site, the South dumping site, and the East dumping site (hereinafter referred to as the North site, the South site, and the East site) were established in 1999, 2006, and 2010, respectively. The restored areas were mainly concentrated in the South site, the East site, and the northwest of the North site. The average restoration intensity in the North site, South site, and East site was 0.515, 0.489, and 0.451, respectively, and the average disturbance intensity was 0.371, 0.398, and 0.320, respectively. The average restoration intensity in the three dumping sites was greater than the average disturbance intensity. This study demonstrates that the combination of temporal remote sensing images and the LandTrendr algorithm can follow the vegetation restoration process of an open-pit mine clearly and can be used to monitor the progress and quality of ecological restoration projects such as vegetation restoration in mining areas. It provides important data and support for accurate ecological restoration in mining areas.

Study of water-conducting fractured zone development law and assessment method in longwall mining of shallow coal seam.

Starting from the source of mining, scientific understanding of surface damage law and assessment method in longwall mining of shallow coal seam is conducive to solving the problems of geological hazards and deterioration of the ecological environment, and promoting the coordinated development of efficient coal mining and environmental protection. Based on numerical simulation and theoretical analysis, the surface damage process and spatiotemporal evolution of fracture field are discussed. The influencing factors and assessment method of surface damage are clarified. The results show that surface damage undergone the immediate roof caving stage, the fracture and instability stage of main roof, the spatial amplification stage of separation layer, the instability stage of surface damage control layer and the mining damage stability stage. Under the critical extraction condition, the cracks above the goaf are divided into the crack area outside the cut, the crack area inside the cut, the re-compaction area in the middle goaf, the crack area behind the longwall face, and the crack area in front of the longwall face. The overburden reaches critical failure ahead of surface critical mining. The sensitivity of loose layer thickness to surface subsidence coefficient is greater than that of mining thickness to surface subsidence coefficient. Surface damage control should be adjusted to local conditions, and finally realize zoning treatment and zoning repair. Through the three-step method of "longwall face rapid advancing method, local grouting reinforcement overburden method and zoning treatment ground fissures method", the surface damage control of 12,401 longwall face is realized. This research provides theoretical guidance and application value for surface ecological restoration in similar mining area.

Absolute bioavailability, dose proportionality, and tissue distribution of rotundic acid in rats based on validated LC-QqQ-MS/MS method.

Rotundic acid (RA), an ursane-type pentacyclic triterpene acid isolated from the dried barks of Ilex rotunda Thunb. (Aquifoliaceae), possesses diverse bioactivities. To further study its pharmacokinetics, a simple and sensitive liquid chromatography with triple quadrupole mass spectrometry (LC-QqQ-MS/MS) method was developed and validated to quantify RA concentration in rat plasma and tissue using etofesalamide as an internal standard (IS). Plasma and tissue samples were subjected to one-step protein precipitation. Chromatographic separation was achieved on a ZORBAX Eclipse XDB-C18 column (4.6 mm × 50 mm, 5 µm) under gradient conditions with eluents of methanol:acetonitrile (1:1, V/V) and 5 mM ammonium formate:methanol (9:1, V/V) at 0.5 mL/min. Multiple reaction monitoring transitions were performed at m/z 487.30 → 437.30 for RA and m/z 256.10 → 227.10 for IS in the negative mode. The developed LC-QqQ-MS/MS method exhibited good linearity (2-500 ng/mL) and was fully validated in accordance with U.S. Food and Drug Administration bioanalytical guidelines. Dose proportionality and bioavailability in rats were determined by comparing pharmacokinetic data after single oral (10, 20, and 40 mg/kg) and intravenous (10 mg/kg) administration of RA. Tissue distribution was studied following oral administration at 20 mg/kg. The results showed that the absolute bioavailability of RA after administration at different doses ranged from 16.1% to 19.4%. RA showed good dose proportionality over a dose range of 10-40 mg/kg. RA was rapidly absorbed in a dose-dependent manner and highly distributed in the liver. In conclusion, this study is the first to systematically elucidate the absorption and distribution characteristics of RA in rats, which can provide additional information for further development and evaluation of RA in drug metabolism and pharmacokinetic studies.

The method of Yiqi Yangyin Tongluo can attenuate the pyroptosis of rheumatoid arthritis chondrocytes through the ASIC1a/NLRP3 signaling pathway.

Background: Based on the ASIC1a/NLRP3 signaling pathway, we explored the specific molecular mechanism of the pyroptosis of rheumatoid arthritis (RA) chondrocytes by the method of nourishing qi, nourishing yin, and dredging collaterals to provide new ideas for the treatment of this disease. Methods: A total of 50 rats were divided into a normal group, model group, methotrexate group, Yiqi Yangyin Tongluo group, and combined group. Except for the normal control group, the other groups used Freund's complete adjuvant (FCA) to make RA rat model. The arthritis index and ankle joint swelling of rats in each group were recorded. HE staining and ELISA were used to assess the pathology of the ankle joint of each group of rats and the content of IL-1ß and IL-18 in rat serum. Furthermore, immunofluorescence and qPCR methods were used to detect the protein and mRNA expression levels of NLRP3, caspase 1, ACS, and ASIC1a in the cartilage tissue of each group of rats. Results: Compared with the normal group, the right hind foot joint of the model group was significantly swollen, the levels of IL-18 and IL-1ß in the serum of rats increased significantly, and the mRNA and protein levels of NLRP3, caspase 1, ACS, and ASIC1a in the chondrocytes also increased significantly. Compared with the model group, the degree of ankle joint swelling and IL-18 and IL-1ß content in rat serum in each medication group was significantly reduced, and the combined group showed the greatest reduction compared with the other groups. After 8 weeks of treatment, compared with the model group, the mRNA and protein levels of NLRP3, caspase 1, ACS, and ASIC1a in the chondrocytes of each medication group were down-regulated. HE staining found that there were large numbers of infiltrating inflammatory cells and pannus in the joint tissue of the model group, while only a small amount of inflammatory cell infiltration and pannus was seen in the joint tissue of the rats in each treatment group. Conclusions: The method of Yiqi Yangyin Tongluo can attenuate the pyroptosis of RA chondrocytes through the ASIC1a/NLRP3 signaling pathway.

Intestinal absorption mechanism of rotundic acid: Involvement of P-gp and OATP2B1.

ETHNOPHARMACOLOGICAL RELEVANCE: Ilicis Rotundae Cortex (IRC), the dried barks of Ilex rotunda Thunb. (Aquifoliaceae), has been used for the prevention or treatment of colds, tonsillitis, dysentery, and gastrointestinal diseases in folk medicine due to its antibacterial and anti-inflammatory effects. However, there is no report about the intestinal absorption of major compounds that support traditional usage. AIM OF STUDY: Considering the potential of rotundic acid (RA) - major biologically active pentacyclic triterpenes found in the IRC, this study was purposed to uncover the oral absorption mechanism of RA using in situ single-pass intestinal perfusion (SPIP) model, in vitro cell models (Caco-2, MDCKII-WT, MDCKII-MDR1, MDCKII-BCRP, and HEK293-OATP2B1 cells) and in vivo pharmacokinetics studies in rats. MATERIALS AND METHODS: The molecular properties (solubility, lipophilicity, and chemical stability) and the effects of principal parameters (time, compound concentrations, pH, paracellular pathway, and the different intestinal segments) were analyzed by liquid chromatography-tandem mass spectrometry. The susceptibility of RA to various inhibitors, such as P-gp inhibitor verapamil, BCRP inhibitor Ko143, OATP 2B1 inhibitor rifampicin, and absorption enhancer EGTA were assessed. RESULTS: RA was a compound with low water solubility (12.89 µg/mL) and strong lipophilicity (LogP = 4.1). RA was considered stable in all media during the SPIP and transport studies. The SPIP and cell experiments showed RA was moderate absorbed in the intestines and exhibited time, concentration, pH, and segment-dependent permeability. In addition, results from the cell model, in situ SPIP model as well as the in vivo pharmacokinetics studies consistently showed that verapamil, rifampicin, and EGTA might have significant effect on the intestinal absorption of RA. CONCLUSION: The mechanisms of intestinal absorption of RA might involve multiple transport pathways, including passive diffusion, the participation of efflux (i.e., P-gp) and influx (i.e., OATP2B1) transporters, and paracellular pathways.

Meta-analysis of randomized clinical trials of adjuvant chemotherapy for resected biliary tract cancers.

BACKGROUND: This meta-analysis was performed by analyzing randomized controlled trials (RCTs) to assess the potential prognostic value of adjuvant chemotherapy (ACT) for patients with resected biliary tract cancers (BTCs). METHODS: PubMed, EMBASE, and the Cochrane Library were searched for relevant articles published. Only RCTs affected by tumors of gallbladder, intrahepatic, perihilar, and distal bile ducts were considered. Data were pooled using a random-effects model. The primary endpoint of the study was overall survival (OS). RESULTS: The study identified 1192 patients who met the inclusion and exclusion criteria. ACT had nearly reached a significant better OS (HR, 0.88; 95% CI, 0.77-1.01; P = 0.07) and achieved a significant better RFS (HR, 0.83; 95% CI, 0.69-0.99; P = 0.04). The effectiveness of ACT for OS was significantly modified by fluorouracil-based ACT (HR, 0.83; 95% CI, 0.70-0.99; P = 0.04), but not by gemcitabine-based ACT (HR, 0.91; 95% CI, 0.74-1.12; P = 0.36). The survival benefit was also not modified by primary disease site, resection margin status, and lymph node status. CONCLUSIONS: ACT is correlated with favorable relapse-free survival compared with non-ACT for resected BTCs patients. Fluorouracil-based ACT could be viewed as a standard practice for resected BTCs patients regardless of the primary cancer site, lymph node or margin status.

[Effects of biological agent dripping on cotton Verticillium wilt and rhizosphere soil microorganism]. / ç”Ÿç‰©è¯å‰‚æ»´æ–½å¯¹æ£‰èŠ±é»„èŽç— åŠæ ¹é™ åœŸå£¤å¾®ç”Ÿç‰©æ•°é‡å’Œå¤šæ ·æ€§çš„å½±å“.

Biological agent is an important green pathway to control Verticillium wilt. A field experiment was performed to examine the effects of Bacillus subtilis agent (15, 30 and 45 kg·hm-2), Trichoderma humatum agent (15, 18 and 24 kg·hm-2), 'Yufeng 99' agent (15, 22.5 and 30 kg·hm-2) and Zhongnonglyukang agent (30, 45 and 60 kg·hm-2) on cotton Verticillium wilt and soil microbial community. The results showed that all of the four biological agents could reduce the incidence and index of cotton Verticillium wilt, with control effects of 50.0%-77.4% in the whole growth period of cotton. The control effects of B. subtilis, 'Yufeng 99' and Zhongnonglyukang agent were positively correlated with the application doses, while that of T. humatum agent of 18 kg·hm-2 was significantly higher than 15 and 24 kg·hm-2. The relative abundance of V. dahliae was significantly reduced, which was negatively correlated with the control effect. The quantity and species richness of soil bacteria were significantly increased with the increases of application doses. The quantity and species abundance of actinomycete were significantly increased, while the quantity of actinomycete varied greatly among different application doses. The quantity and species richness of fungi increased with the increased application doses of T. humatum agent, but was decreased with the increased application doses of other three biological agents, respectively. The control effect was positively correlated with the quantity of bacteria and actinomycetes, but was negatively correlated with that of fungi. Meanwhile, the control effect was significantly and positively correlated with the abundance of Actinomycetes, Nitrospirae, Ascomycota, Chytridiomycota, but was significantly and negatively correlated with that of Deuteromycota. The application doses of 'Yufeng 99', zhongnonglyukang, B. subtilis and T. humatum agents were suggested to be 30, 60, 45 and 18 kg·hm-2, respectively.

Effectiveness of additional resection of the invasive cancer-positive proximal bile duct margin in cases of hilar cholangiocarcinoma.

BACKGROUND: The survival benefits of additional resection of the positive proximal ductal margin (PM) in hilar cholangiocarcinoma (HCCA) remains controversial. This retrospective study investigated the effectiveness of additional resection of the invasive cancer PM under different levels of preoperative carbohydrate antigen 19-9 (CA19-9). METHODS: Patients who underwent hepatectomy for HCCA from 2000 to 2017 were analyzed. Surgical variables, resection margin status, length of the PM (LPM), prognostic factors, and survival were evaluated. RESULTS: A total of 228 patients were enrolled: 175 PM(-) without additional resection patients (group A), 21 PM(-) after additional resection (group B), 16 PM(+) without additional resection (group C), and 16 PM(+) after additional resection (group D). The median survival of group B (20.99 months) was similar to that of group A (23.00 months; P=0.16), and both were significantly better than those of group C (11.60 months) and D (9.50 months), especially when preoperative CA19-9>150 U/mL (P<0.05). The survival of patients with an LPM >10 mm was significantly better compared with those with an LPM ≤10 mm, especially when preoperative CA19-9 was >150 U/mL (P<0.05). Only in the LPM >10 mm group, the survival of group B was comparable with that of group A (P>0.05). CONCLUSIONS: HCCA patients could get a survival benefit from a negative PM resulting from additional resection. Survival could be comparable with that of negative PM without additional resection among HCCA patients. An LPM >10 mm is possibly more associated with better survival compared with whether additional resection of the positive PM is performed under different levels of preoperative CA19-9.

Chinese Guideline on allergen immunotherapy for allergic rhinitis.

The present document is based on a consensus reached by a panel of experts from Chinese Society of Allergy (CSA) and Chinese Allergic Rhinitis Collaborative Research Group (C2AR2G). Allergen immunotherapy (AIT), has increasingly been used as a treatment for allergic rhinitis (AR) globally, as it has been shown to provide a long-term effect in improving nasal and ocular symptoms, reducing medication need, and improving quality of life. AIT is currently the only curative intervention that can potentially modify the immune system in individuals suffering from AR and prevent the development of new sensitization and the progression of disease from AR to asthma. Although the use of AIT is becoming more acceptable in China, to date no AR immunotherapy guideline from China is available for use by the international community. This document has thus been produced and covers the main aspects of AIT undertaken in China; including selection of patients for AIT, the allergen extracts available on the Chinese market, schedules and doses of allergen employed in different routes of AIT, assessment of effect and safety, patients' administration and follow-up, and management of adverse reactions. The Chinese guideline for AR immunotherapy will thus serve as a reference point by doctors, healthcare professionals and organizations involved in the AIT of AR in China. Moreover, this guideline will serve as a source of information for the international community on AIT treatment strategies employed in China.

Primary malignant melanoma of the gallbladder with multiple metastases: A case report.

RATIONAL: Primary malignant melanoma of the gallbladder is an extremely rare tumor, with fewer than 40 cases reported in the literature worldwide. The majority of patients presented as a solitary lesion in the gallbladder. To our knowledge, only one case of primary malignant melanoma of the gallbladder with multiple metastases has been reported, which involved the stomach, duodenum, pancreas, jejunum and a mesenteric lymph node. PATIENT CONCERNS: We report a case of primary malignant melanoma of the gallbladder with metastases to the duodenal bulb, right adrenal and a celiac lymph node. DIAGNOSES: Primary malignant melanoma of the gallbladder with multiple metastases. INTERVENTIONS: Gastrojejunostomy, cholecystectomy, and biopsy of the three metastatic lesions were performed. Histopathologic examination revealed melanin pigments were within the tumor cells of the four lesions, however, junctional activity was noted only in the gallbladder, supporting that the gallbladder was the primary site. No pigmented lesions were detected on the skin or eyes. The postoperative recovery was uneventful, and subsequently, chemotherapy with paclitaxel and carboplatin was administered. OUTCOMES: The patient survived for 16 months due to tumor. progression. LESSONS: The current case was unique due to the adrenal involvement. For patients with multiple metastases of malignant melanoma, gallbladder origin should be considered in the differential diagnosis from cutaneous origin.

[Simultaneous determination of salidroside and tyrosol in Beagle dog plasma using UHPLC-MS/MS after pre-column dansyl chloride derivatization].

A pre-column derivatization method combined with UHPLC-MS/MS was developed for the simultaneous determination of salidroside and tyrosol in Beagle dog plasma. After protein precipitation by acetonitrile, the liquid supernatant was treated with dansyl chloride under dark conditions at 60 ℃ for 30 min, and then, the sample solution was extracted using methyl tertiary butyl ether. The multiple reaction monitoring in positive ion mode was used for MS detection of the tested analytes with the specific ion transitions of m/z 534.2→372.0 for salidroside derivative, m/z 372.0→171.0 for tyrosol derivative and m/z 506.0→171.0 for arbutin derivative. The chromatograph separation was achieved on an ACQUITY UPLC® BEH C18 column (100 mm × 2.1 mm, 1.7 µm) with a gradient mobile phase consisting of acetonitrile (0.1% formic acid)-water (10% acetonitrile, 0.1% formic acid) for 9 min. The assay showed a good linearity over the range of 0.02/0.1 − 20/10 µmol·L−1 with a lower limit of quantitation of 0.02 and 0.1 µmol·L−1 for salidroside and tyrosol in dog plasma, respectively. The intra- and inter-day precisions were all less than 8.68%, and the accuracy was within ±11.4%. The established method with a high sensitivity, good specificity and reliability was appropriate for simultaneous determination of salidroside and tyrosol in dog plasma and successfully applied to a pharmacokinetic study after intragastric administration of salidroside to Beagle dogs.

First case report of retroperitoneal metastasis of fascioliasis after surgery.

RATIONALE: Fascioliasis is a rare cause of liver abscesses, and its clinical course consists of hepatic phase and biliary phase. PATIENT CONCERNS: We describe a 58-year-old female patient who presented with a 2-month history of intermittent fever and abdominal pain. An abdominal computed tomography (CT) revealed confluent low-density lesions in the liver. Complete surgical resection of these abscesses was performed, and postoperative pathological examination and serological tests confirmed a diagnosis of fascioliasis. However, 4 months after the surgery, follow-up CT revealed a lesion in the retroperitoneal area. Meanwhile, ultrasonography-guided percutaneous needle biopsy of the retroperitoneal lesion was performed, and a parasitic infection was suspected. DIAGNOSES: Retroperitoneal metastasis of hepatic phase fascioliasis. INTERVENTIONS: The patient received parasitic resistance treatment with triclabendazole at a dose of 10 mg/kg/d for 2 consecutive days. OUTCOMES: After 2 courses of triclabendazole therapy, the retroperitoneal metastasis regressed to a minor lesion. LESSONS: To the best of our knowledge, this is the first case report of retroperitoneal metastasis of fascioliasis, aimed at helping recognize the clinical features and treatment options of this rare disease.

Expression levels of cytokines and chemokines increase in human peripheral blood mononuclear cells stimulated by activation of the Toll-like receptor 5 pathway.

Recognition of pathogen-associated molecular patterns by Toll-like receptors (TLRs) activates innate and adaptive immune responses. Among the 11 members of the human TLR family, TLR-5 is known to play an important role in the defense against bacterial invasion by binding to flagellin, a conserved component of bacteria. Previous studies have demonstrated that the activation of TLR-5 induces the expression of interleukin (IL)-10, IL-12 and interferon-ß. However, the aim of the present study was to analyze the expression of a wider range of immune-related molecules upon stimulation with a TLR-5 agonist. Following isolation from healthy volunteers, peripheral blood mononuclear cells (PBMCs) were stimulated with flagellin, a TLR-5 agonist. At 4 h after stimulation, quantitative polymerase chain reaction (PCR) and an antibody chip array were conducted to determine the mRNA expression levels of immune molecules and the protein secretion of immune molecules in the supernatant, respectively. The PCR results revealed that activation of TLR-5 significantly influenced the expression of a number of important molecules. In addition, the antibody chip array demonstrated the induction (IL-8) and inhibition [monocyte chemoattractant protein (MCP)-1, MCP-3 and macrophage inflammatory protein-1α) of protein secretion following TLR-5 stimulation. Therefore, the present study demonstrated the importance of TLR-5 in regulating the biological function of PBMCs. In the future, research should focus on the roles of the candidate molecules in TLR-5-mediating functions.

Development of an LC-MS/MS method for the quantitation of deoxyglycychloxazol in rat plasma and its application in pharmacokinetic study.

Deoxyglycychloxazol (TY501) is a glycyrrhetinic acid derivative which exhibits high anti-inflammatory activity and reduced pseudoaldosteronism compared to glycyrrhetinic acid. In this study, a sensitive and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established for the quantitation of TY501 in rat plasma. Plasma samples were treated by precipitating protein with methanol and supernatants were separated by a Symmetry C8 column with the mobile phase consisting of methanol and 10 mM ammonium formate (containing 0.1% of formic acid) (90:10, v/v). The selected reaction monitoring (SRM) transitions were performed at m/z 647.4→191.2 for TY501 and m/z 473.3→143.3 for astragaloside aglycone (IS) in the positive ion mode with atmospheric pressure chemical ionization (APCI) source. Calibration curve was linear over the concentration range of 5-5000 ng/mL. The lower limit of quantification was 5 ng/mL. The mean recovery was over 88%. The intra- and inter-day precisions were lower than 6.0% and 12.8%, respectively, and the accuracy was within ±1.3%. TY501 was stable under usual storage conditions and handling procedure. The validated method has been successfully applied to a pharmacokinetic study after oral administration of TY501 to rats at a dosage of 10 mg/kg.

[The Study of Altered Immune Status of UCMSC upon the Activation of TLR1/2 Pathway].

OBJECTIVE: To study whether the activation of Toll like receptor 1/2 (TLR1/2) has influence on the immune status of umbilical cord mesenchymal stem cell (UCMSC). METHODS: TLR1/2 agonist Pam3CSK4 was conducted in UCMSC-peripheral blood mononuclear cell (PBMC) co-culture system. Flow cytometry and lactate dehydrogenase (LDH) detection were used to measure the proliferation of PBMC and immune attack of PBMC to UCMSC, respectively. The detection of expressions of co-stimulator and stem cell markers of UCMSC upon TLR1/2 activation was completed by flow cytometry. Real-time PCR was introduced to assay the expression of many immune-related molecules in UCMSC. Cell differentiation staining was conducted to study the change of differentiation ability of UCMSC. RESULTS: Activation of TLR1/2 pathway in UCMSC increased the proliferation of PBMC and attack of PBMC to UCMSC, but without influenced the expressions of co-stimulator and stem cell markers. The results of real-time PCR showed that many immune-related molecules were dramatically induced in UCMSC upon TLR1/2 activation, such as Cinterleukin (IL)-2, IL-6, IL-10, IL-12, interferon-γ (IFN-γ), nuclear factor-κB (NF-κB), tumor necrosis factor-α (TNF-α)]. However, the activation of TLR1/2 had no influence on the differentiation ability of UCMSC. CONCLUSION: Activation of TLR1/2 altered the immune status of UCMSC, including increase the immune attack to UCMSC.