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Multi-temporal glacier inventories provide key information about the glaciers, their characteristics, and changes and are inevitable for glacier modelling and investigating geodetic mass changes. However, to date, no consistent multi-temporal glacier inventory for the whole of the Karakoram exists, negatively affecting the monitoring of spatio-temporal variations in glaciers' geometric parameters and their related applications. We used a semi-automatic method combining automatic segmentation and manual correction and produced a multi-temporal Karakoram glacier inventory (KGI) compiled from Landsat TM/ETM+/OLI (Thematic Mapper, Enhanced Thematic Mapper Plus, and Operational Land Imager) images for the 1990s, 2000s, 2010s, and 2020s. Our assessments using independent multiple digitisation of 37 glaciers show that the KGI is sufficiently accurate, with an overall uncertainty of ±3.68 %. We also performed uncertainty evaluation for the contiguous glacier polygons using a buffer of half a pixel, which resulted in an average mapping uncertainty of ±5.21 %. We calculated more than 20 attributes for each glacier, including coordinates, area, supraglacial debris area, date information, and topographic parameters derived from the ASTER GDEM (Advanced Spaceborne Thermal Emission and Reflection Radiometer global digital elevation model). According to KGI-2020s, approximately 10 500 alpine glaciers (>0.01 km2 each) cover an area of 22 510±828 km2 of which 10.18±0.38 % (2290±84 km2) is covered by supraglacial debris. Over the past 3 decades, the glaciers experienced a loss of clean ice and/or snow area but a gain in supraglacial debris. Supraglacial debris cover has increased by 17.63±1.44 % (343.30±27.95 km2), while non-debris-covered glaciers decreased by 1.56±0.24 % (319.85±49.92 km2). The total glacier area was relatively stable and showed only a slight insignificant increase of 23.45±28.85 km2 (0.10±0.13 %). The glacier area has declined by 3.27±0.24 % in the eastern Karakoram, while the glacier area slightly increased in central (0.65±0.10 %) and western Karakoram (1.26±0.11 %). Supraglacial debris has increased over the whole of Karakoram, especially in areas above 4200 m a.s.l. (above sea level), showing an upward shift. The glacier area changes were characterised by strong spatial heterogeneity, influenced by surging and advancing glaciers. However, due to global warming, the glaciers are on average retreating. This is in particular true for small and debris-free glaciers. The multi-temporal KGI data are available at the National Cryosphere Desert Data Center of China: https://doi.org/10.12072/ncdc.glacier.db2386.2022 (F. Xie et al., 2022).

The highly glacierised eastern part of the Tibetan Plateau is the key source region of the five major rivers Yangtze, Yellow, Lancang-Mekong, Nu-Salween and Brahmaputra rivers. These exotic rivers are vital freshwater resources for more than one billion people downstream for their daily life, irrigation, industrial use, and hydropower. However, the glaciers have been receding during the last decades and are projected to further decline which will profoundly impact the water availability of these larger river systems. Although few studies have investigated glacier mass changes in these river basins since the 1970s, they are site and temporal specific and limited by data availability. Hence, knowledge of glacier mass changes is especially lacking for years prior to 2000. We therefore applied digital elevation models (DEMs) derived from large scale topographic maps based on aerial photogrammetry from the 1970s and 1980s and compared them to the SRTM DEM to provide a complete picture of mass change of glaciers in the region. The mass changes are presented on individual glacier bases with a resolution of 30 m and the grided (0.1° and 0.5°). Our database consists of 17444 glaciers with a total area of 17426 ± 523 km2. The annual mean mass loss of glaciers is -0.28 ± 0.15 m w.e. in the whole region. This is larger than the previous site-specific findings, the surface thinning increases on average from west to east along the Himalayas-Hengduan mountains with the largest thinning in the Salween basin. Comparisons between the topographic map-based DEMs and DEMs generated based on Hexagon KH-9 metric camera data for parts in the Himalayas demonstrate that our dataset provides a robust estimation of glacier mass changes. However, the uncertainty is high in high altitudes due to the saturation of aerial photos over low contrast areas like snow surface on a steep terrain. The dataset is well suited for supporting more detailed climatical and hydrological analyses and are available at https://doi.org/10.11888/Cryos.tpdc.272884 (Liu et al., 2022).

This study used remote sensing images from 1996 to 2016 as the main data source, and selected the average annual ecosystem type net change rate, ecosystem type transfer matrix, and comprehensive index of land development degree, to analyze the macro change of the ecosystem pattern in Xishuangbanna Dai Autonomous Prefecture in the past 20 years. Quantitative analysis was performed on amplitude, rate, type of transition, and degree of disturbance of human activities. The results reveal the spatial and temporal changes of the Xishuangbanna ecosystem and their regional differentiation. The results showed that (1) from 1996 to 2016, Xishuangbanna as a whole was dominated by forest ecosystems and rubber ecosystems, followed by tea, farmland, built-up area, and water ecosystems. (2) During 1996-2016, the ecosystem in Xishuangbanna accounted for more than 99% of the total area has not changed. From 1996 to 2003, the transfer of ecosystem types in Xishuangbanna was mainly between forest and rubber ecosystem. (3) The extent of land development and utilization in Xishuangbanna in the past 20 years is relatively low, slightly lower than the national average, and the overall level of land use is at a medium level of utilization, and over time, the degree of disturbance of human activities has shown an increasing trend.

The extremely high proliferation rate of tumor cells contributes to pancreatic cancer (PC) progression. Runt-related transcription factor 1(RUNX1), a key factor in hematopoiesis that was correlated with tumor progression. However, the role of RUNX1 in PC proliferation was still unclear. We found that RUNX1 was significantly upregulated in PC tissues and its expression was negatively associated with prognosis of PC patients in a multicenter analysis according to immunohistochemical (IHC). RUNX1 downregulation in PC resulted in a significantly reduced cell proliferation rate, which was consistent with in vivo subcutaneous tumor formation assay results. RNA-seq and ChIP-seq results revealed that a portion of target genes, including HAP1, GPRC5B, PTPN21, VHL and EN2, were regulated by RUNX1, a finding successfully validated by ChIP-qPCR, qRT-PCR and Western blot. Subsequently, IHC and proliferation assays showed these target genes to be dysregulated in PC, affecting tumor growth. Our data suggest that RUNX1 plays an oncogenic role in tumor proliferation and is a potential prognostic biomarker and therapeutic target for PC.

The Indus River Basin (the Indus) is facing the threat of great water shortages due to rapid population growth, expanding of irrigation area and increasing meltwater from snow and ice under the background of global warming. Being a less gauged basin, the effective usage of water resources in the Indus is always challenged by the high variability of the surface and ground water under a warming climate. This study therefore aimed to characterize and uncover the driving force of changes in water storage in the Indus based on GRACE and GRACE-FO solutions. A series of statistical techniques, such as EOF, modified STL, and Mann-Kenddall test, were applied to quantify and attribute the spatiotemporal patterns of the water storage dynamics. Our results demonstrated that (1) terrestrial water storage anomaly (TWSA) of the basin displayed a deficit and the deficit was largely concentrated in the middle and upper Indus plain (MUIP) during 2002 and 2020. (2) A slight decline in TWSA in the upper Indus basin (UIB) might be attributed to the accelerated melting of glacier and snow cover. (3) The excessive withdrawal of groundwater (1.57 mm/month) dominated the decrease of TWSA over the MUIP although weak increase of precipitation happened in the region. Anthropogenic activities imposed approximately 86.9% impact of the decrease in groundwater and this impact will aggravate for a long time if no effective water management schemes are taken. (4) Influenced by favorable meteorological conditions, the precipitation presented positive trend against the weakness of the India Summer Monsoon and the Westerlies, which exerted the positive influence on TWSA.

Water resources are under severe stress in the highly populated Indus River Basin due to the increased consumption of water across different sectors and climate change. Coping with these challenges, requires a clear understanding on hydrological processes and anthropogenic activities, and how these are influencing recharging and spatiotemporal availability of groundwater in the basin. The present study aims to investigate the natural and anthropogenic impact on Terrestrial Water Storage (TWS) over the Indus River Basin by using a series of statistical methods and the observation data from the Gravity Recovery and Climate Experiment (GRACE) and Follow-On (GRACE-FO). Our results show that (i) TWS Anomaly (TWSA) experienced a significant decrease from 2002 to 2020, particularly in the MUIP; (ii) the UIB showed a weak decreasing trend in TWSA as a result of the accelerated glacier melting; (iii) there was significant loss of groundwater (1.57 mm/month) caused by ineffective water management and over-exploitation; and (iv) assisted by favorable meteorological conditions, the precipitation presented a positive trend against the weakness of the Westerlies, which exerted the positive influence on TWSA.

Previous studies have shown that glacier changes were heterogeneous in the western Karakoram, with the coexistence of retreating, advancing, and surging glaciers. However, it remains unclear that the mechanisms driving these changes. Based on the Shuttle Radar Topography Mission (SRTM) DEM and TerraSAR-X/TanDEM-X images (2014), this study presents glacier surface height changes in the Hunza Basin of the western Karakoram, employing the method of differential synthetic aperture radar interferometry (DInSAR). A slight negative glacier mass balance was observed in the Hunza Basin during 2000–2014. Surge-type glaciers would not have an obvious effect on overall mass balance in regional assessments over long-time scales. Further, glacier surface velocities in the Hunza Basin were estimated from Landsat images for the period of 1990–2018 by utilizing published data sets and Landsat images. Compared to the annual glacier surface velocities, 22 surge events were observed in seven surge-type glaciers in the Hunza Basin. Glacier flow can be attributed to thermally and hydrologically control, and the geomorphological characteristics of different individuals. This study gives us a new insight into the situation of the “Karakoram anomaly” under the background of glacier mass loss in the high mountains of Asia (HMA).

Lake ice, one of the most direct lake physical characteristics affected by climate change, can reflect small-scale environmental changes caused by the atmosphere and hydrology, as well as large-scale climate changes such as global warming. This study uses National Oceanic and Atmospheric Administration, Advanced Very High Resolution Radiometer (NOAA AVHRR), MOD09GQ surface reflectance products, and Landsat surface reflectance Tier 1 products, which comprehensively used RS and GIS technology to study lake ice phenology (LIP) and changes in Qinghai Lake. Over the past 38 years, freeze-up start and freeze-up end dates were gradually delayed by a rate of 0.16 d/a and 0.19 d/a, respectively, with a total delay by 6.08 d and 7.22 d. The dates of break-up start and break-up end showed advancing trends by −0.36 d/a and −0.42 d/a, respectively, which shifted them earlier by 13.68 d and 15.96 d. Overall, ice coverage duration, freeze duration, and complete freeze duration showed decreasing trends of −0.58 d/a, −0.60 d/a, and −0.52 d/a, respectively, and overall decreased by 22.04 d, 22.81 d, and 9.76 d between 1980 and 2018. The spatial pattern in the freeze–thaw of Qinghai Lake can be divided into two areas; the west of the lake area has similar spatial patterns of freezing and ablation, while, in the east of the lake area, freezing and ablation patterns are opposite. Climate factors were closely related to LIP, especially the accumulated freezing degree-day (AFDD) from October to April of the following year. Furthermore, freeze-up start time was more sensitive to changes in wind speed and precipitation.

Xishuangbanna in the southwestern border of China is located in the upper reaches of the Mekong River between the Indo-China Peninsula and the East Asian continent. It is the largest tropical rain forest and monsoon forest in China. They have an irreplaceable ecological service function and are an important barrier to maintaining the ecological security of the Mekong River transboundary basin of the Lancang River. Based on three sets of remote sensing data (1996, 2003, 2010 and 2016), this study has made an exploration on spatio-temporal changes of Xishuangbanna, and also made an assessment on different ecosystem values of six types of ecosystem in Xishuangbanna based on related theories of ecological economics. Results showed that the values of Xishuangbanna ecosystem services in 1996, 2003, 2010, and 2016 were 70.4014billion yuan, 70.2115 billion yuan, 68.5129 billion yuan and 63.6098 billion yuan, respectively. The total value shows a continuous decreasing trend, reflecting the continuous decline in the ability of the Xishuangbanna ecosystem to provide services to humans. If divided by ecosystem types, forest and rubber were two types that have the greatest proportion of service values. In the perspective of service types, soil formation and protection account for the largest proportion, followed by gas regulation and biodiversity protection. Studying Xishuangbanna’s ecosystem service values is helpful to explore the sustainable development of resources and economy.HighlightThis paper used coefficient of sensitivity to determine the dependence of ESV on the change of ecosystem value coefficient over time.The total amount of ESV shows a continuous downward trend, indicating that the ecological environment of Xishuangbanna is still severe.The temporal-spatial change of ESV was analyzed for two decades.

Background: Due to the limitations of strategies for its early diagnosis and treatment, pancreatic cancer (PC) remains a substantial human health threat. We previously discovered a methylation-mediated lncRNA, LINC00261, which is downregulated in PC tissues. However, the underlying role of LINC00261 in PC remains largely unknown. Methods: Quantitative real-time PCR and in situ hybridization were performed to evaluate the expression levels of LINC00261 in PC, adjacent nontumor and normal pancreas tissues. The clinical significance of LINC00261 was assessed in multicenter PC samples. The functions of LINC00261 in PC were investigated by gain- and loss-of-function assays in vitro and in vivo. Potential downstream pathways and mechanisms were explored via RNA sequencing and bioinformatic analyses. RNA immunoprecipitation and chromatin immunoprecipitation assays were used to validate the underlying mechanisms. Pyrosequencing and targeted demethylation of the LINC00261 promoter were performed to explore the upstream epigenetic mechanisms and therapeutic potential. Results: LINC00261 was significantly downregulated in PC tissues, and its expression was positively associated with the prognosis of PC patients. Phenotypic studies indicated that LINC00261 overexpression significantly suppressed PC cell proliferation, migration and metastasis in vitro and in vivo. c-Myc was identified as a downstream target of LINC00261. LINC00261 repressed c-Myc transcription by physically interacting and binding with the bromo domain of p300/CBP, preventing the recruitment of p300/CBP to the promoter region of c-Myc and decreasing the H3K27Ac level. Moreover, the methylation level of the LINC00261 promoter was high in PC tissues and was correlated with poor prognosis. Targeted demethylation of the LINC00261 promoter inhibited PC progression both in vitro and in vivo. Conclusions: Our findings indicate that methylation-mediated LINC00261 suppresses PC progression by epigenetically repressing c-Myc expression. These findings expand the therapeutic potential of LINC00261, possibly providing evidence to support the development of epigenetic drugs or therapeutic strategies. This research adds further insights into the etiology of PC and indicates that LINC00261 may be a prognostic and therapeutic target in PC.

Water supply contributed by glacier and snow melt is of particular significance and play a key role for social stability in the Karakoram region of Upper Indus Basin (UIBKK). Based on the SRTM DEM, TerraSAR-X/TanDEM-X images and ZY3-02 stereo images, the spatiotemporal pattern of glacier mass budgets in the UIBKK have been estimated for the early 21st century (2000–2019) period in this study. A comparison of the total area in 2000 with that in 2019 shows that glacier area remained stable in the early 21st century, with a slight expansion of 56.3 km2 (0.4%). Glaciers in the UIBKK experienced a balanced or slight negative mass budget of −0.08 ± 0.07 m w.e. a−1 in the early 21st century. The spatial variability of mass budget in the UIBKK showed that glacier mass budgets shifted from negative to positive from West to East, respectively, along longitude. Glacier mass loss decelerated from −0.12 ± 0.16 m w.e. a−1 in 2000–2013 to −0.03 ± 0.21 m w.e. a−1 in 2013–2019. No significant differences were observed in thinning rates on debris-covered glaciers (≥19% area covered by debris) and clean-ice glaciers (

Although surging glaciers in the Geladandong Mountain region represent a small percentage of the Qinghai-Tibet Plateau's glacier population, they can provide valuable insight into the glacier processes, flow instabilities, and fast glacier flow. Here we identify 11 surge-type glaciers and reported the observations of surging behaviour in this region by mapping changes in the terminus position, surface velocity, and elevation of glaciers, using a combination of Landsat and ASTER satellite imagery from 1986 to 2020, TerraSAR-X/TanDEM-X, NASADEM, and topographic maps. Among the 11 surge-type glaciers, nine glaciers started to surge in the 1980s and 1990s, and two glaciers began to surge after 2000. In total, we present 22 surface velocity data sets, which show that the active phases usually last 10–19 years and peak surge velocities are between 30 ± 1.26 m a−1 and 100 ± 1.26 m a−1. The acceleration phases at the beginning can last for several years, and the deceleration phases at the end last for a maximum of 2 years, and the flow velocity in summer is greater than that in winter. Under the background of global climate warming, no single process is responsible for glacier surges in the Geladandong Mountain region. The dynamic evolution of glacier surges does not fit neatly within either of the classically cited thermal or hydrological models of surging. We hypothesize that the recent glacier surges may be caused by multiple factors.

Climate warming has a significant impact on the water cycle, especially in regions where the water supply is dominated by snow and glacier meltwater. In this study, spatiotemporal changes of snowpack in the Yarkant River Basin (YRB) and the west of Upper Indus Basin (WUIB), associated with the Karakoram Mountains, were examined using Moderate-resolution Imaging Spectroradiometer (MODIS) daily cloud-free snow cover product. A sensitivity analysis was carried out to explore the climatic effects on snow cover changes. From 2002 to 2018, overall slight (but non-significant) decreasing and increasing trends in snow cover extent were observed in the basins, which occupy the northern and southern slopes of the central and western Karakoram Mountains, respectively. In both basins, the low-altitude areas experienced a slight decrease in snow cover extent, while the high-altitude areas experienced an increase. Significant elevation dependences were found for the snow cover percentage (SCP), snow covered days (SCD) and snow cover melting dates (SCMD), with an inverse elevation dependence for snow cover onset dates (SCOD). Most of the study area showed a decreasing trend in SCD, which was delayed in SCOD, and an advanced trend in SCMD by a rate of

BACKGROUND A large number of studies have shown that KIT mutations are closely related to the prognosis of gastrointestinal stromal tumors (GISTs). At the same time, sunitinib (SU) has become the second-line recommended drug for GISTs because of its efficacy. We initiated a systematic review to compare the efficacy of SU after failure of Imatinib (IM) in different KIT mutations. METHODS We searched for SU-treated patients with advanced GISTs after failed IM treatment by using databases such as PubMed, EMBASE, and the Cochrane Library, up to March 2018. We conducted statistical analyses to calculate the odds ratio (OR), hazard ratio (HR), and 95% confidence interval (CI) using fixed-effects and random-effects models by Review Manager 5.3 software. RESULTS We included a total of 474 patients from 3 retrospective studies and 2 cohort studies. Patients with exon 9 mutations had higher clinical benefit (OR = 2.61, 95% CIs = 1.32-5.18, P = .006) rates and longer progression-free survival (progressive disease, HR = 0.51, 95% CIs = 0.36-0.72, P = .0001) compared with exon 11, but there was no statistically significant difference in overall survival (OS, HR = 0.93, 95% CIs = 0.34-2.55, P = .89) and there was greater heterogeneity (Tau = 0.72, Chi = 21.45, df = 3, P

The preclinical bioanalytical process with animal models begins with sampling biological fluids and tissue. The goal is to understand oral absorption kinetics, distribution, metabolism, excretion, blood brain barrier penetration, drug-drug interactions, and the influences on biomarkers, hematology, electrophysiology, cardiology, blood pressure and behavior. An overview is obtained by periodic blood sampling of 8 - 12 samples over a total time span of 10 - 24h. Urine, feces, bile and microdialysates can augment the information available from whole blood. In today’s preclinical environment, the majority of samples are processed by LC/MSMS augmented by robotic sample preparation tools. These tools save labor and improve precision for smaller volume/lower concentration samples. Our laboratories have been engaged in a project that is focused on improving both the quality and throughput for laboratory animal studies, while providing for reduced numbers of animals and enhanced animal comfort. We have implemented a robotic system that can accomplish most of the above goals for laboratory rats, dogs and primates. Studies with mice are at an earlier stage, but feasibility has been demonstrated. This presentation is a progress report on this evolving research program in cooperation with multiple pharmaceutical and drug development companies. We will illustrate results and discuss future directions.

Daidzin, a soy-derived biologically active natural product, has been reported to inhibit mitochondrial aldehyde dehydrogenase and suppress ethanol intake. This paper describes a method for the determination of daidzin in rat blood. After administration of daidzin, blood samples were periodically collected from awake, freely moving animals by a Culex automated blood sampler. Daidzin was extracted from 50 microl of diluted blood (blood and saline at a ratio of 1:1) with ethyl acetate. Chromatographic separation was achieved within 12 min using a microbore C(18) (100 x 1.0 mm) 3 microm column with a mobile phase containing 20 mM sodium acetate, 0.25 mM EDTA, pH 4.3, 4% methanol and 11% acetonitrile at a flow-rate of 90 microl/min. Detection was attained using a four-channel electrochemical detector with glassy carbon electrodes using oxidation potentials of +1100, 950, 850, 750 mV vs. Ag/AgCl. The limit of detection for daidzin in rat plasma was 5 ng/ml at a signal-to-noise ratio of 3:1. The extraction recovery of daidzin from rat plasma was over 74%. Linearity was obtained for the range of 25-1000 ng/ml. The intra- and inter-assay precisions were in the ranges of 2.7-6.6 and 1.9-3.7%, respectively. This method is suitable to routine in vivo monitoring of daidzin in rat plasma.

A simple gradient reverse-phase liquid chromatographic method has been developed for the separation of daidzin, genistin, daidzein, and genistein in various natural sources and dietary supplements. Peak assignment at various retention times was confirmed by comparison of the simultaneous response of sample peaks to standards, at glassy carbon electrodes held at different oxidation potentials, using a multi-channel electrochemical detector. Identification was substantiated by ESI MS/MS fragmentation patterns. The analytes of interest were extracted by sonicating raw materials in cold 80% methanol/water for 10 minutes. The concentration of the named isoflavones in various dry food and nutritional supplements was determined using the present LCEC procedure.

Due to the lack of mode selection capability, single whispering-gallery-mode (WGM) lasing is a challenge to achieve. In bottle microresonators, the highly nondegenerated WGMs are spatially well-separated along the long-axis direction and provide mode selection according to their axial mode numbers. In this work, we use a loss-engineering approach to suppress the higher-order WGMs and demonstrate single-mode lasing emission in small polymer bottle microresonators. The fiber tapers are not only used to couple pump light into the bottle microresonators to excite the WGMs but also to bring optical losses that are induced from the diameter mismatch between fiber tapers and microresonators. By adjusting the coupling positions, the diameters of fiber tapers, and the coupling angles, single fundamental-mode lasing is efficiently generated with side-mode suppression factors over 15 dB. Our loss-engineering approach is convenient just by moving the fiber taper and may find promising applications in miniature tunable single-mode lasers and sensors.

Single-mode lasing in whispering-gallery mode (WGM) microresonators is challenging to achieve. In bottle microresonators, the highly non-degenerated WGMs are spatially well-separated along the long-axis direction and provide mode-selection capability. In this work, by engineering the pump intensity to modify the spatial gain profiles of bottle microresonators, we demonstrate a simple and general approach to realizing single-mode WGM lasing in polymer bottle microresonators. The pump intensity is engineered into an interference distribution on the bottle microresonator surface. By tuning the spacing between axial positions of the interference pump patterns, the mode intensity profiles of single-bottle WGMs can be spatially overlapped with the interference stripes, intrinsically enabling single-mode lasing and selection. Attractive advantages of the system, including high side-mode suppression factors >20 dB, large spectral tunability >8 nm, low-lasing threshold and reversible control, are presented. Our demonstrated approach may have a variety of promising applications, ranging from tunable single-mode lasing and sensing to nonlinear optics. By engineering the spatial profile of a pump beam, scientists in China have made a bottle-shaped microresonator lase in a single mode. Whispering-gallery-mode microresonators, which rely on light circulating in a closed path around a disk, sphere, toroid or bottle, are popular due to their small size and high quality factor. However, they tend to be multimodal when lasing due to the lack of a mode-selection element. Fuxing Gu at the University of Shanghai for Science and Technology and co-workers overcame this limitation by using a pump beam with a striped interference pattern to excite a single transverse mode of a small polymer bottle microresonator. The result was a miniature single-mode laser with a low lasing threshold and high suppression of unwanted side modes.

Liquid chromatography/tandem mass spectrometry (LC/MS/MS) has been coupled to in vivo microdialysis for on-line monitoring of melatonin in a freely moving rat for a period of 15 hours. A microdialysis probe was surgically implanted into the jugular vein of the rat, and deionized water was used as the perfusion medium at a flow rate of 1.0 microL/min. Microdialysis samples were collected in an on-line injector with sample injection every 30 minutes. Melatonin was dosed by intraperitoneal (i.p.) injection and then monitored by microdialysis/LC/MS/MS. The whole experiment, including the microdialysis sampling and sample injection into the LC/MS system, was fully automated. Metabolites of melatonin were identified off-line by LC/MSn experiments. Two metabolites were identified as 6-hydroxymelatonin and cyclic 2-hydroxymelatonin, consistent with ones found previously in the literature.

Simple and reliable methods for liquid chromatography/electrochemistry (LCEC) and LC/MS/MS with atmospheric pressure chemical ionization (APCI) have been developed for the determination of melatonin in commercially available products. The dissolution of melatonin from the melatonin-containing formulations was simply obtained by dissolving the tablets and capsules in 20% acetonitrile in 0.1 N perchloric acid with ultrasonication. The filtrate of the resultant solution was separated on a 1 × 150 mm C

Tetrahydrobiopterin and seven of its analogues (neopterin, xanthopterin, biopterin, pterin, tetrahydropterin, 6-methyltetrahydropterin, and 6-methylpterin) were separated on a 1 × 150 mm C18 microbore column. These analytes were detected by dual-electrode amperometry and UV absorption. Both a conventional glassy carbon electrode and an interdigitated array microelectrode were used. Low fmol amounts of xanthopterin, tetrahydropterin, tetrahydrobiopterin and 6-methyltetrahydropterin could be determined by electrochemical detection in the oxidative mode, but pmol amounts of the other analogues were determined by electrochemical detection in the reductive mode and with UV detection. Catecholamines and their metabolites do not interfere with the determination of tetrahydrobiopterin and its analogues in biological samples. The developed method was explored for the determination of tetrahydrobiopterin in samples of human urine and rat tissue (brain, liver, and kidney).

Cyclopentenyl cytosine 5′-triphosphate (CPEC-TP) is the active metabolite of the investigational drug cyclopentenyl cytosine (CPEC), a nucleoside analogue which exhibits noteworthy antineoplastic activity against several murine and human tumors in tissue culture, and which is now undergoing Phase I clinical trials. This study describes a method to measure the intracellular CPEC-TP levels in peripheral blood mononuclear cells (PBM cells) from patients treated with CPEC, without using radiolabeled drug. The method utilizes on-line multidimensional high-performance liquid chromatography (HPLC) with two columns of different retention mechanisms connected via an automated programmable switching valve. The elution fraction containing CPEC-TP is initially separated from cellular components using a gel sizing column (TSK-G2000-SW) and then rechromatographed by means of a reversed-phase column operated in an ion-pairing mode (YMC-A-3 12-ODS). The limit of quantitation of CPEC-TP by this method is 2.5 pmol per injection. When CPEC-TP levels were measured in PBM cells from 12 cancer patients after 20 h continuous infusion of CPEC at doses ranging from 3.5 to 5.9 mg/m 2 /h, the levels attained ranged from 1.4 to 13.4 μM (3.6 to 33.5 pmol/10 7 cells). However, wide variability in the concentrations of CPEC-TP achieved were evident at each dose and did not appear to correlate either with the CPEC dose or with CPEC plasma levels. This method was validated by a comparison of the quantitation of CPEC-TP in cultured PBM cells and Molt-4 cells (a human T-cell line adapted for growth in tissue culture) after incubation with both unlabeled and radiolabeled CPEC.

The purpose of the present study was to prepare multivesicular liposomes (MVL) with a high drug loading capacity for intramuscular sustained release and to investigate their potential applicability towards tramadol, and to improve the stability of liposomes by coating PEG. The basic physiochemical properties of tramadol MVLs and PEG-coated MVLs were studied. The average particle sizes of optimum preparation were 18.2 microm and 31.3 microm. The entrapment efficiency was up to 80%. The encapsulation efficiency of tramadol MVLs and PEG-coated MVLs was measured. The results confirmed the possibility of multivesicular liposomes as a sustained-release delivery system. Tramadol was continuously released from MVL formulations in PBS (pH 6.8) in vitro, and reached a maximum of 80% within 72 h. The results show that tramadol PEG-coated MVLs could provide sustained release according to the first order kinetic equation.

The separation and characterization of the metabolic products of lappaconitine in rat urine by high-performance liquid chromatography with electrochemical and ultraviolet detection are described. Urine samples from rats intravenously administered lappaconitine hydrobromide were extracted with chloroform and then purified on a Sep-Pak C18 cartridge. The subsequent resolution into individual compounds was achieved by high-performance liquid chromatography. Identification of these compounds was based on comparisons of the chromatographic behaviour and the detector response with those of authentic samples. Changes in the ratio of lappaconitine to its metabolites in rat urine with time after dosing led to a proposal for one of the probable metabolic pathways of lappaconitine in the rat.

The metabolites of lappaconitine in the urine of humans having been previously administered intramuscularly with lappaconitine hydrobromide were studied using high performance liquid chromatography with electrochemical and ultraviolet detection. The urine was extracted by means of liquid- and solid-phase extractions. Each of the metabolites of lappaconitine was purified by high performance liquid chromatography on a reversed phase column and identified on the basis of the chromatographic behaviour and the detector response. It was proved that lappaconitine, N-deacetyl-16-O-demethyllappaconitine and N-deacetyllappaconitine were excreted in urine from humans receiving lappaconitine.

Cyclopentenyl cytosine (CPE-C) is an investigational drug that is active against human solid tumor xenografts. The 5′-triphosphate of CPE-C inhibits CTP synthase, and depletes CTP and dCTP pools. We conducted a phase I clinical trial of CPE-C given as a 24-h continuous i. v. infusion every 3 weeks in 26 adults with solid tumors. The starting dose rate, 1 mg/m2 per h, was selected on the basis of both preclinical studies and pharmacokinetic data from two patients obtained after a test dose of 24 mg/m2 CPE-C as an i. v. bolus. Dose escalation was guided by clinical toxicity. A total of 87 cycles were given, and ten patients received four or more cycles. The mean CPE-C steady-state plasma levels (Cpss) increased linearly from 0.4 μM to 3.1 μM at dose levels ranging from 1 to 5.9 mg/m2 per h (actual body weight); the mean total body clearance was 146±38 ml/min per m2. CPE-C was eliminated by both renal excretion of intact drug and deamination to cyclopentenyl uracil in an apparent 2∶1 ratio. CTP synthase activity in intact bone marrow mononuclear cells was inhibited by 58% to 100% at 22 h compared to matched pretreatment samples at all CPE-C dose levels. When all data were combined, flux through CTP synthase was decreased by 89.6%±3.1% at 22h (mean ± SE,n=16), and remained inhibited by 67.6%±7.7% (n=10) for at least 24 h post-CPE-C infusion. Granulocyte and platelet toxicities were dose-dependent, and dose-limiting myelosuppression occurred during the initial cycle in two of three patients treated with 5.9 mg/m2 per h. Four of 11 patients (4 of 20 cycles) who received 4.7 mg/m2 per h CPE-C experienced hypotension 24–48 h after completion of the CPE-C infusion during their first (n=2), third (n=1) and sixth cycles (n=1), respectively. Two of these patients died with refractory hypotension despite aggressive hydration and cardiopulmonary resuscitation. One of 12 patients (28 total cycles) treated with 3.5 mg/m2 per h CPE-C experienced orthostatic hypotension during cycle 1, and this patient had a second episode of orthostatic hypotension at a lower dose (3.0 mg/m2per h). Hypotension was not seen in patients receiving ≤2.5 mg/m2 per h CPE-C. The occurrence of hypotension did not directly correlate with either CPE-C Cpss, CPE-U plasma levels, pretreatment cytidine plasma levels, baseline CTP synthase activity, or with the degree of enzyme inhibition during treatment. While the hypotension appeared to be dose-related, its unpredictable occurrence and the uncertainty concerning the mechanism preclude a recommendation of a tolerable dose for future studies.