Spatial distribution dynamics for Epimedium brevicornum Maxim. from 1970 to 2020.

At different time scales, a species will experience diverse distribution changes. For Epimedium brevicornum Maxim, the phenomenon is obvious, but the understanding of the spatial dynamics of E. brevicornum under distinct time scales is poor. In this study, we modeled the potential distribution for E. brevicornum for five time scales, 1970-1979, 1980-1989, 1990-1999, 2000-2009, and 2010-2019, with different occurrence data, and the Kuenm package was used to optimize the parameter combination. Then, SDM tools and a Venn diagram were utilized to simulate the changes in highly suitable areas and spatial dynamics, respectively. Comprehensive results show that temperature seasonality (BIO4, 37.54%) has the greatest effect on the distribution of E. brevicornum, followed by minimum temperature (TMIN, 21.42%). The areas of distribution for E. brevicornum are 35.06 × 105 km2, 25.7 × 105 km2, 67.64 × 105 km2, 27.29 × 105 km2, and 9.87× 105 km2, which are mainly concentrated in Gansu, Shaanxi, Shanxi, and Henan, respectively. In addition, the largest regions for expansion, stability, and contraction under various time scales are 5.6 × 105 km2, 3.54 × 105 km2, and 3.47 × 105 km2, respectively. These changes indicate that approximately 7.96% of the regions are highly stable, and three critical counties, Wanyuan, Chenggu, and Hechuan, and Xixiang, have become significant areas for migration. Overall, our results indicate that there are different spatial distribution patterns and dynamics for E. brevicornum for different time scales. Given these results, this study also proposes comprehensive strategies for the conservation and management of E. brevicornum, which will further improve the current resource utilization status.

Analysis of factors affecting crash under risk scenarios based on driver homogenous clustering.

Research on road safety has focused on analyzing the factors that affect crashes. However, previous studies have often neglected differences in crash causation among heterogeneous clusters of drivers. In particular, the differences in the combined effect mechanisms of the factors in the risk scenarios have not been completely explained. Therefore, this study used the K-means algorithm to perform multidimensional feature homogeneous clustering for drivers involved in crashes and near-crashes. Structural equation modeling involving mediating effects was introduced to explore the direct and indirect effects of each influencing factor on vehicle crashes under risk scenarios and compare the differences in crash causation among different driver clusters. The results indicate that the drivers who experienced the risk scenarios can be classified into two homogeneous driver clusters. Significant differences exist in the demographic characteristics, intrinsic driving characteristics, and crash rates between them. In the risk scenario, traffic factors, distraction state, crash avoidance reaction, and maneuver judgment directly affect the crash outcomes of the two cluster drivers. Demographic characteristics and environmental factors have fewer direct influence on the crash outcomes of two-cluster drivers, but produce more complex mediating effects. Analysis of the differences in the influence of factors between clusters indicates that the fundamental cause of crashes for cluster 1 drivers includes poor driving skills. In contrast, cluster 2 drivers' crashes were more influenced by traffic conditions and their safety awareness. The analysis method of this study can be used to develop more targeted road safety policies to reduce the occurrence of vehicle crashes.

Semantic Segmentation and Depth Estimation Based on Residual Attention Mechanism.

Semantic segmentation and depth estimation are crucial components in the field of autonomous driving for scene understanding. Jointly learning these tasks can lead to a better understanding of scenarios. However, using task-specific networks to extract global features from task-shared networks can be inadequate. To address this issue, we propose a multi-task residual attention network (MTRAN) that consists of a global shared network and two attention networks dedicated to semantic segmentation and depth estimation. The convolutional block attention module is used to highlight the global feature map, and residual connections are added to prevent network degradation problems. To ensure manageable task loss and prevent specific tasks from dominating the training process, we introduce a random-weighted strategy into the impartial multi-task learning method. We conduct experiments to demonstrate the effectiveness of the proposed method.

Predicting the comprehensive geospatial pattern of two ephedrine-type alkaloids for Ephedra sinica in Inner Mongolia.

Ephedra sinica Stapf. is a shrubby plant widely used in traditional Chinese medicine due to its high level of medicinal value, thus, it is in high demand. Ephedrine (E) and pseudoephedrine (PE) are key medicinal components and quality indicators for E. sinica. These two ephedrine-type alkaloids are basic elements that exert the medicinal effect of E. sinica. Recently, indiscriminate destruction and grassland desertification have caused the quantity and quality of these pharmacological plants to degenerate. Predicting potentially suitable habitat for high-quality E. sinica is essential for its future conservation and domestication. In this study, MaxEnt software was utilized to map suitable habitats for E. sinica in Inner Mongolia based on occurrence data and a set of variables related to climate, soil, topography and human impact. The model parametrization was optimized by evaluating alternative combinations of feature classes and values of the regularization multiplier. Second, a geospatial quality model was fitted to relate E and PE contents to the same environmental variables and to predict their spatial patterns across the study area. Outputs from the two models were finally coupled to map areas predicted to have both suitable conditions for E. sinica and high alkaloid content. Our results indicate that E. sinica with high-quality E content was mainly distributed in the Horqin, Ulan Butong and Wulanchabu grasslands. E. sinica with high-quality PE content was primarily found in the Ordos, Wulanchabu and Ulan Butong grasslands. This study provides scientific information for the protection and sustainable utilization of E. sinica. It can also help to control and prevent desertification in Inner Mongolia.

Modeling for Predicting the Potential Geographical Distribution of Three Ephedra Herbs in China.

Ephedra species are beneficial for environmental protection in desert and grassland ecosystems. They have high ecological, medicinal, and economic value. To strengthen the protection of the sustainable development of Ephedra, we used occurrence records of Ephedra sinica Stapf., Ephedra intermedia Schrenk et C.A. Mey., and Ephedra equisetina Bge., combined with climate, soil, and topographic factors to simulate the suitable habitat of three Ephedra based on ensemble models on the Biomod2 platform. The results of the models were tested using AUC, TSS, and kappa coefficients. The results demonstrated that the ensemble model was able to accurately predict the potential distributions of E. sinica, E. intermedia, and E. equisetina. Eastern and central Inner Mongolia, middle and eastern Gansu, and northeastern Xinjiang were the optimum regions for the growth of E. sinica, E. intermedia, and E. equisetina, respectively. Additionally, several key environmental factors had a significant influence on the suitable habitats of the three Ephedra. The key factors affecting the distribution of E. sinica, E. intermedia, and E. equisetina were annual average precipitation, altitude, and vapor pressure, respectively. In conclusion, the results showed that the suitable ranges of the three Ephedra were mainly in Northwest China and that topography and climate were the primary influencing factors.

Priority planting area planning for cash crops under heavy metal pollution and climate change: A case study of Ligusticum chuanxiong Hort.

Introduction: Soil pollution by heavy metals and climate change pose substantial threats to the habitat suitability of cash crops. Discussing the suitability of cash crops in this context is necessary for the conservation and management of species. We developed a comprehensive evaluation system that is universally applicable to all plants stressed by heavy metal pollution. Methods: The MaxEnt model was used to simulate the spatial distribution of Ligusticum chuanxiong Hort within the study area (Sichuan, Shaanxi, and Chongqing) based on current and future climate conditions (RCP2.6, RCP4.5, RCP6.0, and RCP8.5 scenarios). We established the current Cd pollution status in the study area using kriging interpolation and kernel density. Additionally, the three scenarios were used in prediction models to simulate future Cd pollution conditions based on current Cd pollution data. The current and future priority planting areas for L. chuanxiong were determined by overlay analysis, and two levels of results were obtained. Results: The results revealed that the current first- and secondary-priority planting areas for L. chuanxiong were 2.06 ×103 km2 and 1.64 ×104 km2, respectively. Of these areas, the seven primary and twelve secondary counties for current L. chuanxiong cultivation should be given higher priority; these areas include Meishan, Qionglai, Pujiang, and other regions. Furthermore, all the priority zones based on the current and future scenarios were mainly concentrated on the Chengdu Plain, southeastern Sichuan and northern Chongqing. Future planning results indicated that Renshou, Pingwu, Meishan, Qionglai, Pengshan, and other regions are very important for L. chuanxiong planting, and a pessimistic scenario will negatively impact this potential planting. The spatial dynamics of priority areas in 2050 and 2070 clearly fluctuated under different prediction scenarios and were mainly distributed in northern Sichuan and western Chongqing. Discussion: Given these results, taking reasonable measures to replan and manage these areas is necessary. This study provides. not only a useful reference for the protection and cultivation of L. chuanxiong, but also a framework for analyzing other cash crops.

Analyzing Commute Mode Choice Using the LCNL Model in the Post-COVID-19 Era: Evidence from China.

The purpose of this paper is to gain an insight into commuting and travel mode choices in the post-COVID-19 era. The surveys are divided into two waves in Qingdao, China: the first-wave questionnaires were collected under the background of a three-month zero growth of cases; the second wave was implemented after the new confirmed cases of COVID-19. The latent class nested logit (LCNL) model is applied to capture heterogeneous characteristics among the various classes. The results indicate that age, income, household composition, and the frequency of use of travel modes are latent factors that impact users' attitudes toward mass transit and the private car nests when undergoing the shock of the COVID-19 pandemic. Individuals' trepidation regarding health risks began to fade, but this is still a vital consideration in terms of mode choice and the purchase of vehicles. Moreover, economic reinvigoration, the increase in car ownership, and an increase in the desire to purchase a car may result in great challenges for urban traffic networks.

[Planning of ecologically suitable areas for Ligusticum chuanxiong under background of soil cadmium pollution].

Chuanxiong Rhizoma is a traditional Chinese medicinal material mainly produced in Sichuan and Chongqing of China. In recent years, the cadmium content in Chuanxiong Rhizoma produced in most of the genuine producing areas has exceeded the standard, which makes Chuanxiong Rhizoma difficult to be exported. To solve the problem of excessive cadmium content in soil, this study employed the MaxEnt model to simulate the potential geographic distribution of Ligusticum chuanxiong and evaluate important environmental factors, and re-plan its ecologically suitable areas based on the mineral distribution characteristics and soil cadmium pollution status. The results showed that the places suitable for L. chuanxiong growing covered an area of 335 523.69 km~2, mainly in central and eastern Sichuan, southern Shaanxi and most parts of Chongqing. Among them, the highly suitable areas of L. chuanxiong were mainly concentrated in Chengdu, Ya'an, Deyang, and Mianyang. Solar radiation, annual precipitation, and annual range of temperature were evaluated as important variables affecting the distribution of L. chuanxiong, with the contribution rates of 62.3%, 13.3%, and 6.8%, respectively. In addition, Qionglai county, Chongqing county, Mianyang city(Youxian district and Fucheng district), Qingchuan county, and Xinjin county were classified into the first-class ecologically suitable zone, covering a total area of 2 768.87 km~2. The se-cond-class ecologically suitable zone was even wider, involving such counties as Tongjiang county, Renshou county, Jianyang county, and Nanjiang county, and the total area reached 43 616.92 km~2. The re-planning of the ecologically suitable areas for L. chuanxiong has provided strong data support for the cultivation and resource development of L. chuanxiong and also new ideas for solving the problem of excessive cadmium content in L. chuanxiong.

Glucopyranose from Pleurotus geesteranus prevent alcoholic liver diseases by regulating Nrf2/HO-1-TLR4/NF-κB signalling pathways and gut microbiota.

This study investigated the effects of PGPs (Pleurotus geesteranus polysaccharides), a glucopyranose isolated from the mycelium of Pleurotus geesteranus and characterized with the main chain of →4)-α-D-Glcp-(1→, on the prevention against alcohol liver diseases (ALD), with the aim of providing a theoretical basis for the application of P. geesteranus as prebiotic agents in preventing and treating gut dysbiosis and alcohol-related metabolic disorders in individuals with ALD. The results showed that PGP treatment reduced oxidative stress by up-regulating the Nrf2/HO-1 signalling pathways, and decreased the pro-inflammatory factors by down-regulating TLR4/NF-κB signalling pathways. Furthermore, we validated effects of PGPs on balancing the gut-liver axis by maintaining the integrity of the intestinal epithelial barrier of decreasing intestinal permeability, increasing intestinal tight-junction protein and mucin expression and elevating the abundance of short-chain fatty acids (SCFAs) producers in the intestine by regulating the microbiota composition.

Predicting the suitable habitats of parasitic desert species based on a niche model with Haloxylon ammodendron and Cistanche deserticola as examples.

Haloxylon ammodendron, an excellent tree species for sand fixation and afforestation in the desert areas of western China, is threatened by climate change and anthropogenic activities. The suitable habitat of this species is shrinking at a remarkable rate, although conservation measures have been implemented. Cistanche deserticola is an entirely parasitic herb that occurs in deserts, is a source of "desert ginseng" worldwide, and has extremely high medicinal value. Little is known about using niche models to simulate habitat suitability and evaluate important environmental variables related to parasitic species. In this study, we modeled the current suitable habitat of H. ammodendron and C. deserticola by MaxEnt based on occurrence record data of the distributions of these two species in China. We grouped H. ammodendron and C. deserticola into three groups according to the characteristics of parasitic species and modeled them with environmental factors. The results showed that bioclimate was the most important environmental parameter affecting the H. ammodendron and C. deserticola distribution. Precipitations, such as annual precipitation, precipitation seasonality, and precipitation in the driest quarter, were identified as the most critical parameters. The slope, diurnal temperature range, water vapor pressure, ground-frost frequency, and solar radiation also substantially contributed to the distribution of the two species. The proportions of the most suitable areas for Groups 1, 2, and 3 were 1.2%, 1.3%, and 1.7%, respectively, in China. When combined with cultural geography, five hot spot conservation areas were determined within the distribution of H. ammodendron and C. deserticola. The comprehensive analysis indicated that by using MaxEnt to model the suitable habitat of parasitic species, we further improved the accuracy of the prediction and coupled the error of the distribution of a single species. This study provides a useful reference for the protection of H. ammodendron forests and the management of C. deserticola plantations.

Identification of Ligustici Rhizoma et Radix and its adulterants based on their chemical constituents by UHPLC-Q/TOF-MS combined with data mining.

Ligustici Rhizoma et Radix (LR), known as Gaoben (GB) in Chinese, has been used in traditional Chinese medicine for more than 2000 years. However, the varieties of LR are not well characterized at present, and traditional recognition methods have encountered increasing difficulties. This research attempts to distinguish different varieties of LR and its adulterants based on their chemical composition. A total of 61 representative samples were collected, and their varieties were confirmed by combining expert opinion with DNA molecular technology. All of the samples were characterized by an UHPLC-Q/TOF-MS method. The marker components that may contribute to variety discrimination were discerned by a data mining method, and they were then hypothesized, analyzed, and identified. A cluster analysis was performed by partial least squares (PLS) based on their MS signals. Furthermore, a feature extraction to find out the characteristic components, and a correspondence analysis to illustrate the corresponding relationship between the varieties and their components, were developed. As a result, 71 components were identified, of which 27 components were unambiguously identified by comparison with standards. The cluster analysis shows that varieties of LR and its adulterant samples exhibited a certain classification trend, with butylphthalide, senkyunolide I, senkyunolide A, ferulic acid, (Z)-ligustilide, bergapten, levistilide A, vanillic acid, isochlorogenic acid C and isochlorogenic acid A as characteristic chemical components. The varieties and their components showed a modest correlation. In conclusion, our study verified the possibility of discriminating the varieties of LR according to their chemical compositions. This research provides a new reference for the recognition of LR and its adulterants.

A new species of Hypodematium (Hypodematiaceae) from China.

Hypodematium confertivillosum J.X.Li, F.Q.Zhou & X.J.Li, sp. nov., a new species of Hypodematium from Shandong, China, is described and illustrated. It is similar to H. crenatum (Forssk.) Kuhn & Decken and H. glanduloso-pilosum (Tagawa) Ohwi, but differs greatly from them by its abaxial fronds sparsely covered with rod-shaped glandular hairs, its adaxial fronds without rod-shaped glandular hairs and spore reniform, with verrucate processes, surface with distinct finely lamellar rugae ornamentation. The description, photographs and a key to H. confertivillosum as well as their notes are provided.

Geographical classification of Epimedium based on HPLC fingerprint analysis combined with multi-ingredients quantitative analysis.

A reliable and comprehensive method for identifying the origin and assessing the quality of Epimedium has been developed. The method is based on analysis of HPLC fingerprints, combined with similarity analysis, hierarchical cluster analysis (HCA), principal component analysis (PCA) and multi-ingredient quantitative analysis. Nineteen batches of Epimedium, collected from different areas in the western regions of China, were used to establish the fingerprints and 18 peaks were selected for the analysis. Similarity analysis, HCA and PCA all classified the 19 areas into three groups. Simultaneous quantification of the five major bioactive ingredients in the Epimedium samples was also carried out to confirm the consistency of the quality tests. These methods were successfully used to identify the geographical origin of the Epimedium samples and to evaluate their quality.

RNA interference targeting CYP76AH1 in hairy roots of Salvia miltiorrhiza reveals its key role in the biosynthetic pathway of tanshinones.

Plant cytochrome P450s (CYPs) are well known as the largest family of enzymes that contribute to both primary metabolism and the chemical diversity of plant secondary metabolites. It is important to elucidate the in vivo role of CYPs in secondary metabolism, in order to apply them in the production of valuable metabolites in medicinal plants via metabolic engineering. CYP76AH1 has been suggested to catalyze the conversion of the carbon skeleton miltiradiene into the intermediate ferruginol, which is involved in the biosynthesis of tanshinones, the chief bioactive ingredients of Salvia miltiorrhiza. However, its role in planta remains to be elucidated. In this work, we constructed a CYP76AH1 RNAi system for hairy roots. Metabolic analysis of RNAi-AH1 hairy root lines showed a significantly increased accumulation of miltiradiene compared to the control lines. At the same time, the concentration of ferruginol decreased revealing the in vivo catalytic activity of CYP76AH1. The content of tanshinones decreased significantly after silencing of CYP76AH1, which verified its key role in the biosynthesis of tanshinones, and indicated that it could be used as a target for metabolic engineering.

Deep Sequencing of the Scutellaria baicalensis Georgi Transcriptome Reveals Flavonoid Biosynthetic Profiling and Organ-Specific Gene Expression.

Scutellaria baicalensis Georgi has long been used in traditional medicine to treat various such widely varying diseases and has been listed in the Chinese Pharmacopeia, the Japanese Pharmacopeia, the Korean Pharmacopoeia and the European Pharmacopoeia. Flavonoids, especially wogonin, wogonoside, baicalin, and baicalein, are its main functional ingredients with various pharmacological activities. Although pharmaological studies for these flavonoid components have been well conducted, the molecular mechanism of their biosynthesis remains unclear in S. baicalensis. In this study, Illumina/Solexa deep sequencing generated more than 91 million paired-end reads and 49,507 unigenes from S. baicalensis roots, stems, leaves and flowers. More than 70% unigenes were annotated in at least one of the five public databases and 13,627 unigenes were assigned to 3,810 KEGG genes involved in 579 different pathways. 54 unigenes that encode 12 key enzymes involved in the pathway of flavonoid biosynthesis were discovered. One baicalinase and three baicalein 7-O-glucuronosyltransferases genes potentially involved in the transformation between baicalin/wogonoside and baicalein/wogonin were identified. Four candidate 6-hydroxylase genes for the formation of baicalin/baicalein and one candidate 8-O-methyltransferase gene for the biosynthesis of wogonoside/wogonin were also recognized. Our results further support the conclusion that, in S. baicalensis, 3,5,7-trihydroxyflavone was the precursor of the four above compounds. Then, the differential expression models and simple sequence repeats associated with these genes were carefully analyzed. All of these results not only enrich the gene resource but also benefit research into the molecular genetics and functional genomics in S. baicalensis.

Cellular and Subcellular Immunohistochemical Localization and Quantification of Cadmium Ions in Wheat (Triticum aestivum).

The distribution of metallic ions in plant tissues is associated with their toxicity and is important for understanding mechanisms of toxicity tolerance. A quantitative histochemical method can help advance knowledge of cellular and subcellular localization and distribution of heavy metals in plant tissues. An immunohistochemical (IHC) imaging method for cadmium ions (Cd2+) was developed for the first time for the wheat Triticum aestivum grown in Cd2+-fortified soils. Also, 1-(4-Isothiocyanobenzyl)-ethylenediamine-N,N,N,N-tetraacetic acid (ITCB-EDTA) was used to chelate the mobile Cd2+. The ITCB-EDTA/Cd2+ complex was fixed with proteins in situ via the isothiocyano group. A new Cd2+-EDTA specific monoclonal antibody, 4F3B6D9A1, was used to locate the Cd2+-EDTA protein complex. After staining, the fluorescence intensities of sections of Cd2+-positive roots were compared with those of Cd2+-negative roots under a laser confocal scanning microscope, and the location of colloidal gold particles was determined with a transmission electron microscope. The results enable quantification of the Cd2+ content in plant tissues and illustrate Cd2+ translocation and cellular and subcellular responses of T. aestivum to Cd2+ stress. Compared to the conventional metal-S coprecipitation histochemical method, this new IHC method is quantitative, more specific and has less background interference. The subcellular location of Cd2+ was also confirmed with energy-dispersive X-ray microanalysis. The IHC method is suitable for locating and quantifying Cd2+ in plant tissues and can be extended to other heavy metallic ions.

Studies of the microbial metabolism of flavonoids extracted from the leaves of Diospyros kaki by intestinal bacteria.

Flavonoid glycosides are metabolized by intestinal bacteria, giving rise to a wide range of phenolic acids that may exert systemic effects in the body. The microbial metabolism of flavonoids extracted from the leaves of Diospyros kaki (FLDK) by intestinal bacteria was investigated in vitro. High-performance liquid chromatography/linear trap quadrupole orbitrap mass spectrometry was performed to analyze the metabolites of flavonoids in vivo using Xcalibur2.1 software. The results showed that the levels of flavonoid glycosides and flavonoid aglycones decreased rapidly in the process of microbial metabolism by intestinal bacteria in vitro, and the metabolic rate may be related to the concentration of intestinal bacteria in the culture solution. In vivo metabolites of FLDK were detected in rat plasma and urine after oral administration of FLDK. Eight flavonoids were identified in the urine, and three were identified in the plasma; however, flavonoid aglycones were not found in the plasma.

Mucoadhesive microparticles for gastroretentive delivery: preparation, biodistribution and targeting evaluation.

The aim of this research was to prepare and characterize alginate-chitosan mucoadhesive microparticles containing puerarin. The microparticles were prepared by an emulsification-internal gelatin method using a combination of chitosan and Ca2+ as cationic components and alginate as anions. Surface morphology, particle size, drug loading, encapsulation efficiency and swelling ratio, in vitro drug released, in vitro evaluation of mucoadhesiveness and Fluorescence imaging of the gastrointestinal tract were determined. After optimization of the formulation, the encapsulation efficiency was dramatically increased from 70.3% to 99.2%, and a highly swelling ratio was achieved with a change in particle size from 50.3 ± 11.2 µm to 124.7 ± 25.6 µm. In ethanol induced gastric ulcers, administration of puerarin mucoadhesive microparticles at doses of 150 mg/kg, 300 mg/kg, 450 mg/kg and 600 mg/kg body weight prior to ethanol ingestion significantly protected the stomach ulceration. Consequently, significant changes were observed in inflammatory cytokines, such as prostaglandin E2 (PGE2), tumor necrosis factor (TNF-α), interleukin 6 (IL-6), and interleukin1ß (IL-1ß), in stomach tissues compared with the ethanol control group. In conclusion, core-shell type pH-sensitive mucoadhesive microparticles loaded with puerarin could enhance puerarin bioavailability and have the potential to alleviate ethanol-mediated gastric ulcers.

Using a thermoluminescent dosimeter to evaluate the location reliability of the highest-skin dose area detected by treatment planning in radiotherapy for breast cancer.

Acute skin reaction during adjuvant radiotherapy for breast cancer is an inevitable process, and its severity is related to the skin dose. A high-skin dose area can be speculated based on the isodose distribution shown on a treatment planning. To determine whether treatment planning can reflect high-skin dose location, 80 patients were collected and their skin doses in different areas were measured using a thermoluminescent dosimeter to locate the highest-skin dose area in each patient. We determined whether the skin dose is consistent with the highest-dose area estimated by the treatment planning of the same patient. The χ(2) and Fisher exact tests revealed that these 2 methods yielded more consistent results when the highest-dose spots were located in the axillary and breast areas but not in the inframammary area. We suggest that skin doses shown on the treatment planning might be a reliable and simple alternative method for estimating the highest skin doses in some areas.

Biochemical and histopathological effects of subchronic oral exposure of rats to a mixture of five toxic elements.

Arsenic, cadmium, lead, chromium and mercury are toxic elements that occur with high frequency in the soil and thus in agricultural products in China. To explore their combined effects, thirty male and thirty female Sprague-Dawley rats were orally exposed to a mixture of these elements at high dose (2.11, 6.47, 72.82, 2.26 and 2.71 mg/kg bw/day), medium dose (1/10 of the high dose) and low dose (1/100 of the high dose) via their drinking water for 90 days. The results revealed that BUN increased in all rats given the high dose, and serum creatinine increased in female rats given the medium dose. Decreased ALP levels in all treated male rats accompanied decreased RBC counts. In the high dose group, serum albumin, globulin and total protein content decreased in male rats, whereas serum ALT levels increased in female rats. Pathological changes were observed in the hepatocytes and renal tubules of all rats given the high dose, and the numbers of spermatogenic cells and sperm significantly decreased in both the epididymides and testes of the rats given the high dose. Abnormalities were observed earlier and more clearly in the males than in the females under combined exposure.