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.

[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.

Aspartate-modified doxorubicin on its N-terminal increases drug accumulation in LAT1-overexpressing tumors.

L-type amino acid transporter 1 (LAT1), overexpressed on the membrane of various tumor cells, is a potential target for tumor-targeting therapy. This study aimed to develop a LAT1-mediated chemotherapeutic agent. We screened doxorubicin modified by seven different large neutral amino acids. The aspartate-modified doxorubicin (Asp-DOX) showed the highest affinity (Km = 41.423 µmol/L) to LAT1. Aspartate was attached to the N-terminal of DOX by the amide bond with a free carboxyl and a free amino group on the α-carbon atom of the Asp residue. The product Asp-DOX was characterized by HPLC/MS. In vitro, Asp-DOX exerted stronger inhibition on the cancer cells overexpressing LAT1 and the uptake of Asp-DOX was approximately 3.5-fold higher than that of DOX in HepG2 cells. Pharmacokinetic data also showed that Asp-DOX was expressed over a longer circulation time (t1/2 = 49.14 min) in the blood compared to DOX alone (t1/2 = 15.12 min). In HepG2 and HCT116 tumor-bearing mice, Asp-DOX achieved 3.1-fold and 6.4-fold accumulation of drugs in tumor tissue, respectively, than those of the unmodified DOX. More importantly, treatment of tumor-bearing mice with Asp-DOX showed a significantly stronger inhibition of tumor growth than mice treated with free DOX in HepG2 tumor models. Furthermore, after Asp modification, Asp-DOX avoided MDR mediated by P-glycoprotein. These results suggested that the Asp-DOX modified drug may provide a new treatment strategy for tumors that overexpress LAT1 and MDR1.

[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.

Gene expression profiles of various cytokines in mesenchymal stem cells derived from umbilical cord tissue and bone marrow following infection with human cytomegalovirus.

Mesenchymal stem cells (MSCs) have both multi-lineage differentiation potential and immunosuppressive properties, making them ideal candidates for regenerative medicine. However, their immunosuppressive properties potentially increase the risk of cancer progression and opportunistic infections. In this study, MSCs isolated from human umbilical cord blood (UCMSCs) and adult bone marrow (BMMSCs) were infected with human cytomegalovirus (HCMV). Cytopathic changes were observed 10 days post infection. PCR products amplified from genomic DNA and cDNA were used to confirm the HCMV infection of the UCMSCs and BMMSCs. Real-time PCR was conducted to quantify the expression of immunomodulatory molecules, including cytokines, chemokines, growth factors, adhesion molecules and cancer-related genes. Our results indicate high upregulation of the majority of these molecules, including many growth factors, tumor necrosis factor alpha, interleukin-8, interleukin-6 and interferon gamma. Adhesion molecules (VCAM-1, TCAM-1 and selectin-E) were downregulated in the infected UCMSCs and BMMSCs. Antibody chip array evaluation of cell culture media indicated that the growth factor secretion by UCMSCs and BMMSCs was greatly influenced (p < 0.001) by HCMV. The stimulation of MSCs with HCMV led to the activation of downstream signaling pathways, including pSTAT3 and Wnt2. Our results show that HCMV can significantly alter the functions of both UCMSCs and BMMSCs, although not in the same way or to the same extent. In both cases, there was an increase in the expression of proangiogenic factors in the microenvironment following HMCV infection. The discrepancy between the two cell types may be explained by their different developmental origin, although further analysis is necessary. Future studies should decipher the underlying mechanism by which HCMV controls MSCs, which may lead to the development of new therapeutic treatments.

Does internal stenting for pancreaticojejunostomy decrease the rate of pancreatic fistula following pancreatic resections? A meta-analysis.

BACKGROUND/AIMS: The aim of this study is to evaluate the effectiveness of using an internal stent for pancreaticojejunostomy (PJ) on pancreatic fistula (PF) formation, as well as on the overall outcome for patients undergoing pancreatic resections. METHODOLOGY: Articles published until the end of February 2012 comparing internal stenting and no stenting for PI were included. The primary outcome of interest was PF. The secondary outcome of interest included operative time, intra-operative blood loss, overall morbidity, hospital mortality and postoperative length of hospital stay. RESULTS: Five articles were identified for inclusion. The meta-analysis revealed that internal stenting for PJ was not associated with a statistically significant reduction in PF rate (OR 1.03; 95% CI=0.70 to 1.51; p=0.88). Patients with soft pancreas had higher PF rate in stenting group, but the difference was not significant (OR=1.71; 95% CI=0.95 to 3.10; p=0.08). There was no significant difference between the two groups in operative time, intra-operative blood loss, overall morbidity, hospital mortality, and postoperative length of hospital stay. CONCLUSIONS: The current literature suggests that internal stenting for PJ following pancreatic resections does not decrease the rate of pancreatic fistula or alter overall patient's outcome.

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.

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.

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.

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.

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.

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.

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.

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.

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.

The protective mechanism of ligustrazine against renal ischemia/reperfusion injury.

BACKGROUND: Ischemia/reperfusion (I/R) injury is unavoidable in renal transplantation, and represents an additional risk factor for the late renal allograft failure. Our study focused on the effects of ligustrazine on oxidative stress, apoptosis, neutrophils recruitment, the expression of proinflammatory mediators and adhesion molecules caused by renal I/R injury. MATERIALS AND METHODS: Renal warm I/R was induced in male C57BL/6 mice by clamping the left renal artery and vein non-traumatically. Group I was sham-operated animals; group II, nontreated animals; and group III, ligustrazine-treated animals (80 mg/kg, i.p. 30 min before I/R). Mice were sacrificed 4 and 24h post reperfusion. The effects of ligustrazine on oxidative stress, neutrophils recruitment, proinflammatory mediators, and adhesion molecules caused by renal I/R injury were assayed. RESULTS: Ligustrazine pretreatment attenuated dramatically the injuries in mice kidneys caused by warm I/R (histological scores of untreated versus treated, 4.2 ± 0.4 versus 0.9 ± 0.3; P<0.01). Administration of ligustrazine significantly reduced myeloperoxidase (MPO) activity by 38.6% and decreased malondialdehye (MDA) level by 19.2%, while superoxide dismutase (SOD) activity increased by 39.6% (P<0.01), suggesting an effective reduction of oxidative stress following ligustrazine treatment. Moreover, ligustrazine also inhibited cell apoptosis, abrogated neutrophils recruitment, and suppressed the over expression of TNF-α and ICAM-1. CONCLUSIONS: In conclusion, ligustrazine protects murine kidney from warm ischemia/reperfusion injury, probably via reducing oxidative stress, inhibiting cell apoptosis, decreasing neutrophils infiltration, and suppressing the overexpression of TNF-α and ICAM-1 levels.

Epidermal growth factor receptor as a target for anti-proliferative treatment of proliferative cholangitis in hepatolithiasis.

BACKGROUND: In recent years, with a deeper understanding of pathologic changes in hepatolithiasis, more and more attention has been paid to the relationship of postoperative remnant proliferative cholangitis (PC) with stone recurrence and biliary restenosis, but effective management strategies have not yet been developed. Thus, the aim of this study was to determine whether epidermal growth factor receptor inhibitor (AG-1478) could inhibit hyperplasia and lithogenic potentiality of PC. METHODS: The PC animal model was established via retrograde insertion of a 5-0 nylon thread into the common bile duct through Vater's papilla. The common bile duct in the therapeutic group received a single intraluminal administration of AG-1478, followed by weekly intraperitoneal injections of AG-1478. Subsequently, influence of EGFR inhibitor on hyperplasia, apoptosis, and lithogenic potential of PC were evaluated via histology, expression changes of EGFR, BrdU, Ki-67, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), Fas, mucin 5 AC, and collagen I. RESULTS: EGFR inhibitor AG-1478 was effective not only in inhibiting the mRNA and protein expression of EGFR, BrdU, and Ki-67, but also in increasing Fas mRNA expression and TUNEL-positive cells, as a result leading to the inhibition of hyperplasia of the biliary epithelium, submucosal gland, and collagen fibers in the diseased bile duct. Additionally, collagen I expression and fibrous thickness of the bile duct wall was significantly reduced, thereby reducing the incidence of biliary tract stricture secondary to PC. Also of note, treatment with AG-1478 could efficiently decrease the lithogenic potential of PC via inhibition of mucin 5AC expression and mucoglycoprotein secretion, hereby facilitating prevention of stone recurrence. CONCLUSION: EGFR antagonist AG-1478 had a potent anti-proliferative and anti-fibrotic effectiveness on PC and, therefore, holds promise as a candidate of PC treatment.

Rapamycin in combination with donor-specific CD4+CD25+Treg cells amplified in vitro might be realize the immune tolerance in clinical organ transplantation.

It is an urgent need to induce and keep the donor-specific immune tolerance without affecting the function of normal immune defense and immune surveillance in clinical organ transplantation. Large number of studies showed that both the establishment of donor-recipient chimerism and the application of antibodies or drugs could obtain the donor-specific immune tolerance in animal transplantation model. However, the former as treatment of clinical practice has a poor feasibility, the latter has a very low success rate in clinical organ transplantation. There is a group of naturally occurring CD4(+)CD25(+) regulatory T cells (Tregs) that mediate immune tolerance by suppressing alloreactive T cells in vivo. These cells are unable to curb the occurrence of allograft rejection owing their low content. And donor-specific Tregs amplified in vitro alone can not induce donor-specific immune tolerance for recipient. Rapamycin (RPM) as a proliferation signal inhibitor, studies have shown it can effectively inhibit allograft rejection and maybe contribute to induction of immune tolerance. But there exist still many dose-dependent adverse reactions which could prevent the establishment of immune tolerance and reduce the life quality of recipients in the clinical application of RPM. Therefore, we speculate a small amount of RPM combined with donor-specific Tregs amplified in vitro may be not only induce the achievement of donor-specific tolerance, but also reduce or eliminate the side effects of RPM in clinical organ transplantation.

Combination of rapamycin and IL-2 do not affect antigen presentation ability of rat B cell and could promote Tregs proliferation and inhibitory activity.

Rapamycin (RPM), a powerful agent used clinically in transplant recipients, induces CD4(+)CD25(+) regulatory T cells (Tregs) which play an important role in induction of immune tolerance. However, long-term use of RPM has negative side effects. In this report, we found that combination with the low dose RPM and high dose IL-2 did not affect antigen presentation of rat B cells to Tregs, and could efficiently promote Tregs proliferation and enhance their inhibitory activities in vitro. In addition, the combination of low dose RPM and high dose IL-2 enhanced mRNA expression of Foxp3, TGF-beta1 and Pim-2 in Tregs but not in CD4(+)CD25(-) T effector cells (Teffs). The Tregs inhibitory activity is positively associated with mRNA expressions of TGF-beta1 and Pim-2 while unrelated to the Foxp3 mRNA expression. Our present study offers one approach to expand functional Tregs in vitro, which maybe used for clinical immune tolerance induction.

Liquid chromatographic/mass spectrometry assay of bromotetrandrine in rat plasma and its application to pharmacokinetic study.

A rapid and sensitive liquid chromatography-tandem mass spectrometric method (LC-MS/MS) for the determination of bromotetrandrine in rat plasma has been developed and applied to pharmacokinetic study in Sprague-Dawley (SD) rats after a single oral administration. Sample preparation involves a liquid-liquid extraction with n-hexane-dichlormethane (65:35, containing 1% 2-propanol isopropyl alcohol, v/v). Bromotetrandrine and brodimoprim (internal standard, IS) were well separated by LC with a Dikma C18 column using methanol-ammonium formate aqueous solution (20 mm) containing 0.5% formic acid (60:40, v/v) as mobile phase. Detection was performed on a triple quadrupole mass spectrometer in multiple reaction monitoring mode. The ionization was optimized using ESI+ and selectivity was achieved using MS/MS analysis, m/z 703.0 --> 461.0 and m/z 339.0 --> 281.0 for bromotetrandrine and IS, respectively. The present method exhibited good linearity over the concentration range of 20-5000 ng/mL for bromotetrandrine in rat plasma with a lower limit of quantification of 20 ng/mL. The intra- and inter-day precisions were 2.8-7.5% and 3.2-8.1%, and the intra- and inter-day accuracy ranged from -4.8 to 8.2% and -5.6 to 6.2%, respectively. The method was successfully applied to a pharmacokinetic study after a single oral administration to SD rats with bromotetrandrine of 50 mg/kg.