As a targeted drug， monoclonal antibodies have been successful in tumor therapy. Thus， antibodies and related products are the fastest-developing biological agents. There are currently more than 40 monoclonal antibodies in the market that have been approved by the FDA， half of which are used to treat cancer. In recent years， a new generation of antibody drugs， such as human antibody， glyco-engineered antibody， bispecific antibody， antibody-drug conjugates and immune checkpoint blockade antibody， have successfully cured various malignant tumors. In this paper， the history of antibody treatment for cancer， and the development and prospect of anti-tumor antibodies have been reviewed.

Recently， increasing  cancer  researches  focus on  antibody-drug  conjugates （ADCs） which can improve the anti-tumor  potency  with  less  adverse  effect  while  benefiting  patients  in the future. However， safety evaluation of ADCs is a big challenge because of complex components as well as in experience in preclinical studies. In this review， the authors reviewed the mode of action， hazard risks， and toxicity observed in preclinical/clinical studies of ADCs， summarized the preclinical studies of Adcetris（brentuximab vedotin） and Kadcyla（ado-trastuzumab emtansine）， and suggested a better strategy of ADCs in preclinical safety evaluation.

There is an increasing demand for neonatal and juvenile animal toxicity studies during the research and development of new drugs. In this paper， we discussed general evaluation parameters of pediatric non-clinical safety with pediatric drugs， such as growth and development and food intake， and paramenters of other organs and systems, such as the central nervous system， reproductive system，
behavior evaluation in combination with our own experience.  In addition， the characteristics of non-clinical safety evaluation of new traditional Chinese medicine materia medica used for juvenile animals were analyzed. This paper is intended reference for non-clinical safety evaluation of pediatric drugs and to gain some experience related to formulation of new guidelines.

Drug toxicology，as a discipline which studies drug toxicity mechanisms and evaluates comprehensive drug safety is of crucial importance for guiding sensible clinical drug use，reducing adverse drug reactions and reducing failures of new drug development caused by toxicity. This article summarizes the basic situation of Natural Science Foundation of China（NSFC）funded projects in drug toxicology between 2001 and 2015，involving the amount of approved grants，funding rate，funding category and supported faculty，and the change of NSFC guidelines. Research topics，ideas and contents of NSFC funded projects are generalized and characteristics，problems and future trends are also analyzed to provide reference for research on drug toxicology.

There have been increasing reports on liver injury induced by traditional Chinese medicine（TCM）and natural medicine in recent years. The risk of liver injury induced by TCMs has attracted more attention at home and abroad. In this paper，epidemiologic characteristics，risk factors，clinical features of TCMs induced liver injury were discussed. The myth about research and safety evaluation of TCMs induced liver injury was analyzed. Based on the property of TCMs and characteristics of their clinical use，this paper proposed the priorities of basic and clinical safety evaluation of TCMs induced liver toxicity. It is hoped that this study may provide reference for scientific evaluation of liver toxicity of TCMs.

OBJECTIVE Dynamics of serum and urine metabolites in hepatic injury rats induced by ethanolic extract from Rhizoma Dioscoreae Bulbiferae（RDB） was investigated by 1H-NMR-based metabonomic methods in order to discover early biomarkers of liver toxicity induced by RDB. METHODS Rats were ig adminisetred with RDB at a dose of 5 g·kg-1 for 28 d. Rats were sacrificed 3，7，14 and 28 d after RDB administration，as well as after a recovery period，respectively. Blood was taken for routine biochemical analysis by an automatic biochemical analyzer. Liver/body mass indexes were calculated，and liver pathological changes were observed with hematoxylin-eosin staining. Urine samples were collected before and 3，7，14 and 28 d after RDB administration，respectively，as well as after withdrawal. Metabonomic analysis was carried out for serum and urine samples. Principal component analysis and orthogonal partial least squares- discriminant analysis were used for screening and identifiying early biomarkers.RESULTS Compared with the control group，total bilirubin（TB）and total cholesterol（TC）values were increased in 3-28 d in RDB group（P<0.05， P<0.01） . Total bile acid（TBA）was elevated in 7-28 d（P<0.05， P<0.01） . TB， TC and TBA became normal after discontinuation with RDB. There was no significant difference between RBD- treated group and control group in the activity of glutamic- pyruvic
transaminase and glutamic-oxaloacetic transaminase，and the content of glucose also was not different between the two groups. The ratio of liver/body mass was elevated at 3-28 d（P<0.01）but returned to normal after withdraval of RDB. The enlargement and necrosis of hepatocytes were observed 7 d after RDB administration，and lesion degree was aggravated with the extension of RDB delivery time. Metabonomic analysis showed that the serum lipids（low density lipoprotein/very low density lipoprotein（LDL/VLDL），glutamic acid，choline phosphate and glycerolphosphatecholine were increased in the early stage. Pyruvate and N- acetylglutamate were decreased in urine. These metabolites became normal 7 d after discontinuation with RDB. CONCLUSION The serum lipids（LDL/VLDL），glutamic acid，glycerol phosphate choline，as well as urine pyruvic acid salt and N- acetyl glutamate may be used as the early biomarkers for liver toxicity induced by RDB.

Type 2 diabetes mellitus（T2DM）is a complicated endocrine disease caused by bothgenetic and environmental factors，many of which are unknown as far as for the pathogenesis ofT2DM is concerned. In recent years，diabetes researchers have paid more attention to the intestinal microbiota. The discovery , verification and molecular mechanism of the intestinal microbiota related to T2DM are the focus of this article. The role of intestinal microecology in the development of T2DM is also reviewed，involving diet，chronic inflammation，short chain fatty acids，bile acids，the intestinal microbiota and circadian rhythm，and microRNA.

OBJECTIVE To study the antidepressant effects of ammoxetine（AMX）and the underlying mechanisms. METHODS Two behavioral despair models，the tail suspension test（TST） and the forced swimming test（FST），were used to evaluate the antidepressant-like effects of AMX 2.5-20 mg·kg-1 following oral administration. Monoamine neurotransmitter p-chloro-phenylalanine（p-CPA）and α-methyl-ptyrosine（AMPT）depletion models in mice were used to investigate the effects of AMX on levels of 5-serotomin（5-HT）and norepinephrine（NE）in the brain. RESULLTS The results of behavioral study showed that compared with normal control group，AMX（10 and 20 mg·kg- 1）reduced the immobility time of mice by 51.4% and 80.7% in the TST（P<0.05，P<0.01）or by 48.0% and 66.2% in the FST（P<0.05），respectively. Locomotion activity test indicated that AMX did not increase or decrease the movement distance of mice，demonstrating that AMX had no excitatory or inhibitory actions on the central nervous system. Moreover，AMX（5， 10 and 20 mg·kg-1）exerted antidepressant effects in the p-CPA induced 5-HT depletion model and AMPT induced NE depletion model，as evidenced by the significantly reduced immobility time，ie，63.9%，93.4%，90.5% and 61.9%，77.2%，100% reduction in the TST （P<0.01），respectively，and AMX at the dose of 20 mg·kg-1 significantly increased the concentrations of 5- HT and NE by 144.7% and 57.2% in the mouse brain（P<0.05），respectively. CONCLUSION AMX has strong antidepressant-like effects in behavioral despair models and monoamine neurotransmitter depletion models in mice，which is involved in the increased levels of 5-HT and NE in the brain.

Abstract: Ubiquitin- specific proteases (USPs) regulate the deconjugation of ubiquitin tag from substrate proteins. Dysregulation of USPs has been linked to many diseases. Despite extensive efforts by academia and industry，high quality chemical molecules targeting the USPs family of ubiquitin enzymes are still scarce. In this commentary，I propose the criteria of a high quality chemical probe and the strategies to target USPs.

OBJECTIVE To examine the reversal effect of desipramine (DMI) on resistance to temozolomide(TMZ) in U251/TR cells and explore its mechanism. METHODS U251/TR cells were exposed to DMI (20-80 μmol•L-1) or TMZ (0.5-10 mmol•L-1) for 24 h, cell viability was determined by cell counting kit-8 assay with IC50 calculated. The cytotoxicity of U251/TR cells treated with TMZ (1 or 2 mmol•L-1) in combination with DMI (20, 30 or 40 μmol • L- 1) for 24 h was detected using CCK- 8 assay. Synergism between DMI and TMZ was analyzed by the JIN Zheng-jun method. Apoptosis of U251/TR cells induced by TMZ 1 mmol • L- 1, DMI 30 μmol • L- 1,or their combination was examined by Hoechst33258 stains and caspase 3 activity was detected by luminescence analysis. Expression of C/EBP homologous protein (CHOP) was measured using quantitative real- time PCR and Western blotting. The survival rate of U251/TR cells treated with TMZ 1 mmol•L-1 and/or DMI 30 μmol•L-1 was also assessed after silencing CHOP expression by small interference RNA (siRNA). RESULTS DMI or TMZ alone inhibited the growth of U251/TR cells significantly in a concentration- dependent manner (r 2=0.983,0.982,P<0.05), with the IC50 (33.6± 0.5)μmol•L-1 and (2.5 ± 0.6)mmol•L-1, respectively. The cell viability inhibitory rate of U251/TR cells by TMZ (1 or 2 mmol•L-1) combined with DMI (20, 30, or 40 μmol•L-1) was greater than that by TMZ or DMI alone (P<0.05). The JIN Zheng-jun analysis revealed that combination of DMI and TMZ produced synergistic cytotoxicity (Q>1.15), ie, compared with TMZ alone, TMZ (1 mmol•L-1) com⁃ bined with DMI (30 μmol•L- 1) produced significant nuclear fragmentation and condensation (P< 0.05). In addition, DMI and TMZ in combination activated caspase 3 activity in U251/TR cells (P<0.05). Knock⁃ down of CHOP by specific siRNA attenuated the synergistic effect of DMI in the presence of TMZ, the survival rate of the combined drug group raised from 51.8% to 62.2%(P<0.05). CONCLUSION The results suggest that DMI reverse resistance of U251/TR cells to TMZ through activation of the CHOP-depend⁃ ently apoptosis pathway.

Abstract: Spontaneous rhythmic activity of pacemaker neurons in the central nervous system underlies fundamental neurological processes such as locomotion, cognition and circadian rhythm. Among the wide range of ion channels required for its generation, the Ca2+-activated K+ (KCa) channels play a prominent role in maintaining physiologically-relevant frequency and pattern of pacemaker activity. Much of our understanding of the functions of KCa channels in pacemaker neurons have been derived from pharmacological studies using channel modulators, such as iberiotoxin and apamin. Despite the significant advances made, recent studies have painted an increasingly complex picture of the effects of widely used KCa channel modulators on unintended targets that may confound our under⁃ standing of their functions. In this review, we discussed the utility and shortcomings of the KCa channel modulators, and highlighted the significance of these findings, because the KCa channel modulators have been used in early clinical trials to treat disorders ranging from Parkinson disease to alcoholism.

Abstract: Calcium signaling is a critical component of neuronal development, and is mediated by numerous calcium channels, exchangers and calcium- binding proteins. Growth cones use calcium gradient as a tool to pathfind correctly, ultimately directing the appropriate growth of axons. Consequently, dysfunctions of calcium channels could lead to incorrect pathfinding, thus resulting in developmental disorders. It is important to understand the pharmacological modulators of the calcium channels present in the growth cone-some already widely used in other tissue pathologies, like cardiovascular disease. Further, L-type channel blockers such as dihydropyridines have been instrumental in understanding the properties of L-type channels as well as their heterogeneous nature. Investigation of channel property often uses pharmacological inhibition to reveal their functionality. Recent studies show the channel modulators as promising drug targets in the treatment of neurodevelopmental disorders. The goal of this article is to discuss the potential effects of pharmacological regulators of growth cone calcium channels on outgrowth activity.

Abstract: Neurodegenerative diseases refer to incurable conditions that result in progressive degeneration or death of nerve cells. This causes functional movement deficits and cognitive problems such as dementias. Among different types of neurodegenerative diseases, Alzheimer disease (AD) accounts for majority of the cases. The transient receptor potential melastatin member 2 (TRPM2) channel is a Ca2+-permeable non-selective cation channel which has been studied and implicated in the pathological process of AD through different pathways including inflammation. This review summarizes the contribution of TRPM2 in AD pathology and recent advances in pharmacology of TRPM2, with a focus on relationships between β- amyloid, oxidative stress and Ca2 + . We also discuss the potential future research direction for neurodegenerative diseases.

Abstract: Mitochondrial dynamic is very essential biological process in mammalian cells. In healthy cells，mitochondrial dynamic keeps a balance between fission and fusion process. Dynamic related protein 1 (Drp1) plays an essential role in mitochondrial division and is the key point of the distribution and function of mitochondria in axons，dendrites and synapses. Studies in patients with Alzheimer disease suggested that the dysfunction or abnormal expression of Drp1 makes the balance of mitochondrial dynamic fission and fusion in Alzheimer disease moved to the harmful side in hippocampus cells，leading to the imbalance of dynamic changes. Thus，Drp1 may be a valuable and novel target of mitochondrial dynamic changes for treatment of memory loss associated with Alzheimer disease.

Abstract: Drug addiction is a chronic，relapsing brain disorder，which develops，in part，because of aberrant learning and memory. Accumulative studies during recent decades demonstrated that addictive drug hijacks the normal memory circuit in the brain to form a long-lasting drug reward memory，which determines relapse to addictive drug. In this review，we will describe what has been learned about drug reward memory，especially focused on one of the associative drug reward memory models，druginduced conditioned place preference. Drug reward memory is a dynamic process，which consists of several stages，including acquisition，consolidation，maintenance，retrieval，reconsolidation and extinction. Interventions with pharmacological in these memory processes will differentially regulate drug reward memory. Furthermore，the recently developed novel pure behavioral procedure according to the hypothesis of memory processes，e.g. post-retrieval extinction，could erase drug reward memory，which shows more advantages than the pharmacological medications that used in memory studies. Finally， we discussed two major methodological issues in drug reward memory，procedure and timing，which should be carefully considered when designing the related studies and interpreting the results from related studies. So far，it is not sure whether it is feasible to develop a pharmacological medication that only erases drug reward memory without impairing normal memories，we propose that inhibition of drug reward memory would be a good strategy to limit the risk of relapse to addictive drug. Although current findings on drug reward memory benefits little for treatment of drug addiction，the ongoing studies on drug reward memory will provide a promising strategy for reducing the risk of relapse to addictive drug.  

Abstract: There are more than two dozens of peptide hormones that are produced and released from the gastrointestinal (GI) tract. Among them, the incretin hormone glucagon-like peptide 1 (GLP-1) has received the most intensive attention for the past 30 years. Functional studies on GLP-1 and anoth⁃ er gut incretin hormone glucose-dependent insulinotropic peptide (GIP) have led to the development of novel diabetes therapeutic agents known as GLP-1 receptor agonists and DPP-Ⅵ inhibitors. Instead of forming endocrine glands, the gut hormone producing endocrine cells are widely spread throughout the entire GI tract, permitting vital interactions with the ″external″ environment. Here a brief introduction on GLP-1 and how nutritional components regulate its secretion were made, followed by reviewing some key development on how gut environment affects the production and secretion of GLP-1, including the contribution of gut microbiota.

Since entering the 21st century，the trend of high incidence of chronic diseases in the in⁃ ternational community has not been effectively controlled and the pharmaceutical and medical fields are put forward new challenges. Both the development and evaluation of chemical drugs，biological drugs or traditional Chinese medicine are facing with how to significantly improve the efficacy and reduce the side effect of the drugs. With the coming of the accurate medical treatment as well as the big data era，new opportunities and challenges are needed either in the development of new drugs or new use of old drugs. However，as known that the human body consists of the symbiotic microbial groups of human microecological system especially the gut microecology system is more and more important in human health and disease，and even part diseases such as obesity and diabetes have a certain degree of causality with the gut microflora，so we need to reanalyze the core ideas about drug development or even the current medical theory system. Therefore，it is possible to step out of the old way of current drug development in the new period，to make new contributions on the real effective implementation of prevention and control of chronic diseases. According to international human body microecological especially the research progress of gut microecological field，systematic analysis of the problems existing in the current research and development of new drugs，will not only focus on the human body itself，but also the human microecology as well as the balance between human and micro⁃ ecological，which will promote rapid development of research and development of new drugs and prevention and control of chronic disease in order to improve the physical and mental health services in our country.

OBJECTIVE To investigate the effect of amifostine（Amf）on the differentiation of human megakaryocyte cell line-Dami. METHODS Dami cells were treated with Amf 0.01-5.0 mmol•L-1 for 12 d. Dami cells were counted every day for the growth curve：only cells with a diameter >20 μm. The platelet demarcation membrane system was observed by transmission electron microscopy. The expression of CD33，CD34，CD41a and DNA ploidy was detected by flow cytometry. RESULTS Amf 0.1-1.0 mmol•L-1 promoted the differentiation of Dami cells，but inhibited their proliferation at a concentration >1.0 mmol•L-1. When these cells were treated with Amf 1.0 mmol•L- 1 for 12 d，the platelet demarcation membrane system was observed，the percentage of cells with a diameter >20 μm was increased by 24.6%（P< 0.01），the expression of CD41a was increased by 11.9%，while the expression of CD33 was decreased by 13.6%（P<0.05） . Polyploidy cells（16N）were observed，and 4N， 8N and 16N cells were increased to 31.56%，8.83% and 3.43%，respectively（P<0.05） . CONCLUSION Amf 0.1-1.0 mmol•L- 1 can promote the differentiation of Dami cells，but inhibit their proliferation at a high concentration（>1.0 mmol•L-1）.

Abstract：Ambient fine particulate（PM2.5）pollution has become one of the major environmental issues that endanger human health in China，and how to recognize and prevent its health hazards is one of current hot spots for scientific research. Ambient PM2.5 has maintained high levels over recent years，and accurate assessment of personal exposure to PM2.5 is the basis for in-depth investigation. Ambient PM2.5 contains various chemical constituents from multiple pollution sources，and can exert a series of short-term and long-term health hazards via a range of biological mechanisms. Epidemiological and toxicological studies are two major types of design used in the health hazards assessment of PM2.5. In addition，interventional studies on how to mitigate the adverse health effects of PM2.5 are emerging as a new important sphere for future scientific research.

OBJECTIVE To discuss the effect of hepatotoxicity of water extraction of Dioscorea bulbifera L.（WED）and anti-oxidative mechanism of mitochondrial adenosine triphosphate（ATP）and superoxide dismutase（SOD） . METHODS Thirty- six male ICR mice were divided into four groups： normal control（water），and WED 19.6，28.0 and 40.0 g•kg-1（ig）for 14 d，before the activity of serum alkaline phosphatase（ALP）， glutamic-pyruvic transaminase（GPT）and glutamic-oxaloacetic transferase （GOT） was detected. The content of ATP，reactive oxygen species （ROS） and malondialdehyde （MDA）and the activity of total SOD in liver tissue of mice were detected，so were the activities of Na+- K +-ATPase， Ca2 +-Mg2 +-ATPase and Mn2 +-SOD in mitochondria of liver tissue. RESULTS Compared with normal control group，liver indexes of WED 28.0 and 40.0 g•kg-1 groups were increased（P<0.01） . The activity of serum GPT，GOT and ALP in WED 40.0 g•kg-1 group was increased（P<0.05），but the content of ATP in WED 19.6，28.0 and 40.0 g•kg-1 groups was decreased（P<0.01） . The content of MDA and ROS in WED 40.0 g•kg-1 group was increased（P<0.05） . The activity of Na+-K+-ATPase and Ca2+-Mg2+- ATPase in WED 40.0 g•kg- 1 group was decreased in mitochondria of liver tissue（P<0.05），but the activity of Mn2 +- SOD was increased（P<0.05） . CONCLUSION WED may induce liver injury and the mechanism is possibly related to mitochondria injury by weakening ATP synthesis and oxidative damage.

Abstract： OBJECTIVE To explore the possibility that Dichroa alkali salt（DAS）can enhance efficacy and reduce toxicity when combined with artemisinin and its derivatives. METHODS Kunming mice were used in acute toxicity test of DAS. Diarrhea rate，mortality，50% lethal dose（LD50），LD90，50% diarrhea dose（DD50）and DD90 were determined，and the safe dose of DAS was defined. An animal model of malaria was established by inoculating strains of the parasite in an antimalarial in vivo experiment. The model mice were ig given drugs，once a day，for 4 d. The titer was compared between alone use and combined use of DAS with artemisinin and its derivatives. Blood of the caudal vein was dynamically collected for blood smear. Gemsa dying assay was used to observe whether the change of the parasite and its revival under a microscope in each group. Finally，negative conversion ratio，anti- revival ratio，50% effective dose（ED50） and ED90 were calculated. RESULTS The safe dose of DAS was 0.5 mg•kg- 1 in acute toxicity test. While a low dose of DAS（0.25 and 0.5 mg•kg- 1）was alone used， negative conversion ratio was relatively low and anti-revival effect was not obvious. In vivo experiments of anti-mouse malaria indicated that compared with alone use of DAS 0.5 mg•kg- 1 or artemisinin with its derivatives〔artemisinin，dihydroartemisinin（Dih），artemether and artesunate〕，negative conversion ratio and anti-revival ratio were increased significantly（P<0.01， P<0.05）when DAS 0.5 mg•kg-1 and artemisinin（134，192，274，392 and 560 mg•kg-1），Dih（52， 80 and 123 mg•kg-1），artemether（10， 20， 40 and 80 mg•kg-1）and artesunate（32.5， 65.0 and 130.0 mg•kg- 1）were used in combination. When DAS 0.5 mg • kg- 1 and artemisinin with its four derivatives were in combination used，the titer was increased 1.8，1.2，2.0 and 1.6 times compared with alone use of artemisinin and its derivatives， respectively. In addition，diarrhea or death didn′t occur in combined groups. CONCLUSION DAS has a narrow safe dose window. DAS in combination with artemisinin and its derivatives can possibly reduce toxicity while enhancing efficacy.

The seminar on novel ideas and methods in pharmacological researches，and new drug research and development of traditional Chinese medicine（TCM），organized by the Professional Committee of Pharmacology on Traditional Chinese Medicine and Natural Medicine， Chinese Pharmacology Society，was held in Tengzhou，Shandong Province，on August 5，2016. Professor ZHANG Yong-xiang，chair of the committee，presided over the seminar. Professor LIU Jian-xun and LI Lin delivered keynote speeches. More than 30 members of the committee from all over the country attended the seminar. The participants had a broad and in-depth discussion on issues concerning phar⁃ macological researches and new drug research and development of TCM. The ideas and proposals by some committee members were summarized，hoping to provide reference in the pharmacological researches and new drug research and development of TCM.

OBJECTIVE To investigate the effect of clopidogrel（Clog），a platelet aggregation inhibitor，on the development of colitis-associated colon cancer（CAC）and its possible mechanism. METHODS To establish a CAC model，male BALB/c mice were treated with single azoxymethane（AOM） 10 mg·kg- 1 by ip. One week later，the mice drank 2.5% dextran sulfate sodium（DSS）for one week and water for two weeks，which lasted three cycles. From the first day mice received 2.5% DSS water， Clog 12.5，25.0 and 50.0 mg·kg-1 was ig administered once a day. Body mass，clinical symptoms，the number of colon tumor and tumor size in colon tissue were recorded. Hyperplasia of tumors was analyzed by HE staining. In the early inflammatory phase of the CAC model，the length of colons was measured， histological structure and epithelium cell proliferation of colon tissues were evaluated by HE staining and Ki67 staining，respectively. In the tumorigenesis and progression phase of the CAC model，epithe⁃ lium cell proliferation of colon tissues was evaluated by Ki67 staining. The mRNA expression of tumor necrosis factor-α（TNF-α）was detected by real-time quantitative PCR. The expression of chemokine（C-X-C motif）ligand 2（CXCL2）and its receptor 2（CXCR2）in colon tissues was detected by PCR and immu⁃ nohistochemistry. RESULTS Compared with model group，clinical symptoms of mice in Clog 12.5 mg·kg-1 group were alleviated，the size of colon tumors was decreased（P<0.05），and hyperplasia of tumors was reduced（P<0.05） . During the inflammatory phase，the clinical symptoms of mice in Clog 12.5 mg·kg-1 group were significantly alleviated（P<0.05），the decrease of body mass was reduced（P<0.01），the colon shrinkage was ameliorated（P<0.01）， the inflammatory injury and epithelium cell proliferation in colon tissues were reduced（P<0.05） . During the tumorigenesis and progression phase，epithelium cell prolif⁃ eration in colon tissues in Clog 12.5 mg·kg-1 group was reduced（P<0.01），and the mRNA and protein expression of TNF- α，CXCL2 and CXCR2 of colon tissues was decreased（P<0.05） . CONCLUSION Clog can alleviate inflammation during the CAC early inflammatory phase and inhibit the formation of CAC. The antitumor effect of Clog may be related to the decrease in expression of CXCL2 and CXCR2.

Most of the toxic herbs have never been subjected to pharmacokinetic or toxicokinetic studies so far, and content testing has never become mandatory. To make matters worse, there is no reference substance available for these toxic components. Therefore, we should encourage and strengthen basic scientific research and guide the establishment and validation of the methodology for pharmacokinetics or toxicokinetics of the toxic herbs so as to improve the risk management of the traditional Chinese materia medica compound preparations of toxic herbs. This paper is intended to introduce the concept of toxic herbs and their toxic ingredients and effects, focusing on the toxicokinetics of the traditional Chinese materia medica compound preparations including toxic herbs.

Alzheimer disease (AD), the most common dementia, is a chronic, progressive and neuro-degenerative disorder. With an increasing prevalence, AD has been the third cause of death after cardiovascular diseases and cancer in the elderly population. However, the pathogenesis of AD remains unclear, which has led to a fairly slow development of drugs for AD and a dim view of future treatments of AD. It has been a hot spot and a big challenge to develop effective, therapeutic drugs for AD. Recently, this topic was discussed via WeChat by experts from the Neuropsychiatric WeChat Group, which consists of 300 Chinese-origin neuroscientists and neuropsychiatrists in China or overseas. The experts pointed out the problems that might have misled researches on drug discovery, such as the misleading but dominating AD pathology hypotheses and problems with the platforms for drug screening. Therefore, it is important to review the pathology of AD and the treatment strategies from big data and the overall view of the disease, which may shed new light on AD therapy to develop drugs for multiple targets, leading to omni-directional, comprehensive treatments of AD. The development of AD can be further classified into different stages based on the upstream factors of AD pathology. Interestingly, it has been found that the AD brain has mitochondria damage and dysfunction; long-term exposure to low doses of ionizing radiation can also cause AD-like pathological changes. These provide novel views and ideas in terms of the pathological process and preventive and therapeutic strategies for AD.

OBJECTIVE  To make a comparison of sensitivity and specificity between serum miR-122 and traditional biomarkers of drug-induced liver injury. METHODS Acetaminophen (APAP, 1250 mg•kg^-1,ig), α- naphthylisothiocyanate (ANIT, 150 mg • kg^-1, ig), methionine-choline deficient diet (MCDD, free feeding) and carbon tetrachloride (CCl₄, 50%, ip) were used to establish hepatocellular injury, cholestasis,steatosis and fibrosis models, respectively, which were used to evaluate the sensitivity of serum miR-122 as a biomarker of drug-induced liver injury, when compared with the traditional biomarkers. Isoprenaline hydrochloride (IH) and gentamicin (GM) were used to establish myocardial and renal injury models, respectively, which were used to evaluate the specificity of serum miR-122, when compared with the traditional biomarkers. Serum and liver tissues were collected at different time points during the study. Traditional biomarkers such as glutamic- pyruvic transaminase (GPT), glutamic-oxaloacetic transaminase (GOT) and total bilirubin (TBIL) were measured with an automatic biochemistry analyzer. MiR-122 was detected with real- time quantitative PCR. Histopathological examination with HE staining was performed for liver tissues. RESULTS Serum miR-122 increased significantly earlier [miR-122 was significantly increased (>2 fold) at 1.5 h, 3 h, 2 weeks and 4 weeks after treatment in the four models respectively, while no significant increase (<2 fold) was observed for GPT, GOT and TBIL at 6 h, 12 h, 3 weeks and 6 weeks after treatment in the four models respectively] and more signficantly (the maximum fold change for miR-122 was 235.8, 202.7, 73.8 and 600.3 for the four models, respectively. For the GPT, the maximum fold change was 9.5, 3.9, 2.5 and 6.6, respectively; 6.0, 2.4, 1.4 and 3.5 respectively for GOT; 2.6, 2.3, 1.2 and 1.8 respectively for TBIL) than that of traditional biomarkers in hepatocellular injury, cholestasis, steatosis and fibrosis models, when compared with the control group. In the myocardial injury model, a significant increase of GOT was observed after IH treatment (2.1 fold), while no change was observed for serum miR-122. In the renal injury model, no false positive results were observed for serum miR-122. CONCLUSION  Serum miR-122 can be used as a biomarker of drug-induced liver injury and serum miR-122 is more sensitive and specific than traditional biomarkers (such as GPT, GOT and TBIL).

The active ingredients in some Tibetan medicinal herbs are toxic components as well, and we need to have a clear understanding of their mechanism and metabolic pathways in use. The endogenous toxic components of highly toxic Tibetan herbal medicines are mainly alkaloids, such as aconitum alkaloids, methyllycaconitine, tropane alkaloids, brucine, strychnine, papaverine and swainsonine. The majority of endogenous toxic alkaloids in Tibetan medicine herbs exist in roots, fruits and seeds of plants, exerting neurotoxicity or cardiotoxicity as highly toxic inherent chemicals. Most alkaloids are metabolized in phaseⅠvia de-alkylation, hydroxylation, hydrolysis and other reactions, as well as in phaseⅡ via glucuronic acid and sulfonic acid conjugation. They form various metabolites with high polarities and reduced toxicities so as to be easily excreted. The closeness between the therapeutic dose and toxic dose of alkaloids components in Tibetan medicinal herbs leads to their attenuated preparation via frying, dairy, highland barley wine soaking, or in combination with Terminalia Chebula to decrease toxicity, as is cited classic books on in Tebitan medicine. Focused on twelve alkaloids of five classes including aconitine, tropane and brucine, we have reviewed the characteristics of their metabolism and transformation, as well as their toxicity attenuation and safety evaluation.

2019年是中华人民共和国建国70 周年，为总结展示建国以来我国药理学研究及创新药物研发的成果，及时交流新成就和新经验，并不断提高药理学研究、新药研发和临床合理用药的水平，向中华人民共和国成立70周年献礼，经中国药理学会第十一届常务理事会研究决定，于2019年11月10-13日在北京市召开“中国药理学会第十五次全国学术大会”。本次会议邀请国际药理学联合会（IUPHAR）领导及国内外知名专家、院士和国家有关部门领导到会做大会特邀报告，并组织专题学术论坛和青年英文报告。专题交流主要包括临床药理学、心血管药理学、药物代谢、网络药理学、海洋药物药理学、生殖药理学、教学与科普、肾脏药理学、民族药物生化与分子机制研究及综合交叉药理学等。

2019年是中华人民共和国建国70 周年，为总结展示建国以来我国药理学研究及创新药物研发的成果，及时交流新成就和新经验，并不断提高药理学研究、新药研发和临床合理用药的水平，向中华人民共和国成立70周年献礼，经中国药理学会第十一届常务理事会研究决定，于2019年11月10-13日在北京市召开“中国药理学会第十五次全国学术大会”。本次会议邀请国际药理学联合会（IUPHAR）领导及国内外知名专家、院士和国家有关部门领导到会做大会特邀报告，并组织专题学术论坛和青年英文报告。专题交流主要包括临床药理学、心血管药理学、药物代谢、网络药理学、海洋药物药理学、生殖药理学、教学与科普、肾脏药理学、民族药物生化与分子机制研究及综合交叉药理学等。

2019年是中华人民共和国建国70 周年，为总结展示建国以来我国药理学研究及创新药物研发的成果，及时交流新成就和新经验，并不断提高药理学研究、新药研发和临床合理用药的水平，向中华人民共和国成立70周年献礼，经中国药理学会第十一届常务理事会研究决定，于2019年11月10-13日在北京市召开“中国药理学会第十五次全国学术大会”。本次会议邀请国际药理学联合会（IUPHAR）领导及国内外知名专家、院士和国家有关部门领导到会做大会特邀报告，并组织专题学术论坛和青年英文报告。专题交流主要包括临床药理学、心血管药理学、药物代谢、网络药理学、海洋药物药理学、生殖药理学、教学与科普、肾脏药理学、民族药物生化与分子机制研究及综合交叉药理学等。

OBJECTIVE  To construct lentiviral vectors overexpressing wild type and enzymatic activity deficiency heparanase (HPA) in order to find out more about the enzymatic effects and non-enzymatic effects of HPA. METHODS  Primer sequences for seamless cloning were designed according to the vector sequence and gene sequence (wild type and enzymatic activity mutant: E225/343A). Then, linearized vector and genes amplified by PCR were recombinated. The highly purified recombinant plasmids were obtained by transformation, PCR identification, sequencing identification and plasmid extraction. The constructed expressing plasmids and packaging plasmids were co-transfected into HEK-293T cells, and cell culture supernates were collected and concentrated to obtain lentiviral infection solutions. HEL cells were infected with the virus and the expression of HPA was detected by Western blotting. The enzymatic activity of overexpressed HPA (wild type and enzymatic activity mutant) was validated by detecting the amounts of heparan sulfate (HS) on the cell surface using flow cytometry, while the amounts of syndecan-1 in cell culture supernates were detected using ELISA. RESULTS  The sequencing results of recombinant plasmids were completely the same as expectated, and the overexpression of HPA was validated by Western blotting. As for enzymatic activity, flow cytometry results revealed that the Geometry mean fluorescence intensity of cell surface HS in HEL cells overexpressing wild type HPA (160.0±8.0) was significantly lower than that of control cells (345.0±15.2), whereas its syndecan-1 levels in cell culture supernates (59.0 ±3.8) ng·L-1 were higher than those of control cells (32.2±3.9) ng·L-1. However, there was no significant difference in the amounts of HS (353.0±14.0 vs 345.0±15.2) or syndencan-1 ［(30.9±2.9) ng·L-1 vs (32.2±3.9) ng·L-1)］ between HEL cells overexpressing E225/343A mutated HPA and control cells. CONCLUSION  Lentiviral vectors overexpressing wild type and enzymatic activity mutant HPA have been constructed and validated in HEL cell line, providing a novel HPA gene-editing tool for studying the physiological and pathological effects of HPA.

OBJECTIVE  To establish a preparation system for megakaryocytes (MKs) derived from human embryonic stem cells (hESCs) and MK microparticles (MKMPs), and to assess the pro-angiogenic efficiency of these microparticles. METHODS  ① hESCs were induced to mesodermal progenitor cells via monolayer culture with the first-stage induction medium for 2 days before the cells were induced to hemogenic endothelial/hematopoietic progenitor cells by culturing with the second-stage 
induction medium for another 3 days. Then, the cells were dissociated into single cells, seeded into the third-stage induction medium, and cultured using the suspension method for 8 days to obtain MKs. The specific characters of differentiated cells were identified through morphological observation and flow cytometry before stage-specific marker proteins in different periods were analyzed [hESCs: TRA-1-60, sialyl glycolipid stage-specific embryonic antigen4 (SSEA4)];mesodermal progenitor cells: brachyury; hemogenic endothelial/hematopoietic progenitor cells: CD34, CD43; MKs:CD41a, CD42b), and immunofluorescence staining [β1-tubulin, von Willebrand factor (VWF)], [friend leukemia integration 1 (FLI1), CD42]. ② MKMP collection and verification: MKMPs were collected via differential centrifugation. The concentration and size of these MKMPs were determined by nanoparticle tracking analysis (NTA), and both the morphology and ultrastructure were examined by transmission electron microscopy (TEM). Besides, the MKMPs-specific proteins [CD41, tumor susceptibility gene 101 (TSG101) and CD9] were detected by Western blotting analysis. ③ Biological function of MKMPs: MKMPs were stained with CD41a-PE antibodies and co-cultured with human umbilical veinvascular endothelial cells (HUVECs) labeled by CD34-APC for 3 h. Live-cell immunofluorescence was employed to find out whether HUVECs could absorb MKMPs. To find out whether MKMPs could affect the role of HUVECs in angiogenesis and cell migration, platelet microvesicles (PMPs) were used as positive controls. The experimental groups were added with different concentrations of microparticles (1, 5, 10 and 20 mg·L-1) while the control group was given no microparticles (0 mg·L^-1). The number of nodes that formed the lumen after 5 h of incubation in Matrigel was counted, and the size of healing of the scratch area was analyzed after 6 h. To elucidate the mechanism through which MKMPs impacted angiogenesis, ELISA was used out to quantitatively detect the concentration of proteins in microparticles. RESULTS  ① A three-stage differentiation cultural system was established to develop hESCs into MKs. Flow cytometry revealed progressive loss of pluripotency markers SSEA4 and TRA-1-60, while the mesodermal 
progenitor marker brachyury peaked at d 2. Subsequently, hemogenic endothelial/hematopoietic progenitor markers CD34 and CD43 emerged at d 5, followed by megakaryocytic markers CD41a and CD42b at d 13. Immunofluorescent images further demonstrated that MKs expressed specific proteins CD42, β1-tubulin, von VWF and FLI1 at d 13. ② Microparticles were collected via differential centrifugation. Transmission electron microscopy revealed that their substructure exhibited a typical double-layered membrane. Nanoparticle tracking analysis indicated that the size was (164.3±14.0) nm. The result of WB demonstrated that the microparticles expressed specific markers, including TSG101, CD9 and CD41. ③ MKMPs were absorbed after being co-cultured with HUVECs for 3 h and enhanced the ability of HUVECs to form tubes and migrate. Notably, the treatment of 5 mg·L-1 MKMPs was more effective than 5 mg·L-1 PMPs treatment. The results of ELISA showed that the content of VEGF from MKMPs was higher than from PMPs, which may be the key factor in regulating endothelial biological function. CONCLUSION  MKs derived from hESCs can generate functional microparticles which can promote angiogenesis.

OBJECTIVE  To investigate the mechanisms of adrenal dysfunction among adult offspring rats caused by prenatal ethanol exposure (PEE) aggravated by high-fat diet (HFD) and unpredictable chronic stress (UCS). METHODS  Pregnant rats were randomly divided into the control group (saline, ig, once) and PEE group (4 g·kg^-1, ig, once) from gestational day (GD) 11 until delivery. At postnatal week 4 (PW4), offspring rats from the control group and PEE group were randomly assigned to three subgroups: the normal diet (ND) group, HFD group, and HFD-UCS group (n=20 per subgroup, at an equal male-to-female ratio). The rats in each group were given a corresponding diet until PW24. The HFD-UCS group received HFD until PW24, with additional UCS treatment initiated at PW21 and continued for 3 weeks. Serum adrenocorticotropic hormone (ACTH) levels were measured by radioimmunoassay, and corticosterone (CORT) levels were quantified via enzyme-linked immunosorbent assay (ELISA). Adrenal mRNA expressions of steroidogenic factor 1 (Sf1), steroidogenic acute regulatory protein (Star ), cytochrome P450 cholesterol side chain cleavage(P450scc), 3β-hydroxysteroid dehydrogenase (3β-Hsd), steroid 21-hydroxylase (P450c21), steroid 11β-hydroxylase (P450c11), 11β-hydroxysteroid dehydrogenase type 1 (11β-Hsd1), 11β-Hsd 2, mineralocorticoid receptor (Mr ), glucocorticoid receptor (Gr ), insulin-like growth factor 1 (Igf1), IGF1 receptor (Igf1r), and serine/threonine kinase 1 (Akt1) were determined using real-time quantitative PCR. Protein expressions of Akt1 and phosphorylated Akt1 (p-Akt1) were analyzed via immunohistochemistry. RESULTS  In male offspring, HFD/PEE significantly reduced serum ACTH and CORT levels, downregulated Star, P450scc, 3β-Hsd and P450c11 mRNA, decreased 
11β-Hsd1 mRNA expressions and 11β-Hsd1/11β-Hsd2 ratio, but increased Igf1, Igf1r mRNA, and p-Akt1 protein. Conversely, HFD-UCS/PEE elevated ACTH, CORT, P450scc, 3β-Hsd and P450c11 mRNA, increased 11β-Hsd1 and 11β-Hsd1/11β-Hsd2 ratio while suppressing Igf1 mRNA, and p-Akt1 protein. In females, HFD/PEE decreased ACTH but upregulated Igf1, Akt1 mRNA, and p-Akt1 protein. HFD-UCS/PEE increased ACTH, CORT, Sf1, Star, P450scc and P450c11 mRNA, and 11β-Hsd1/11β-Hsd2 ratio, but reduced lgf1 and Igf1r mRNA. CONCLUSION  HFD/UCS can aggravate PEE-induced adrenal dysfunction in adult offspring. The adrenal "GC-IGF1 axis" is an endocrine negative feedback one, and its decompensation may increase the susceptibility of a wide range of GC-related adult chronic diseases, such as diabetes.

Commonly used anesthetic sedatives (opioids, benzodiazepines, ketamine, propofol, etc.) share the risk of inducing respiratory depression, and their multi-target mechanism of action presents significant heterogeneity. Opioids inhibit the rhythmic activity of the respiratory center of the medulla bulbar (such as the PreBötzinger complex and parbrachial nucleus) by activating both the μ-opioid receptor and the G-protein-gated inwardly-rectifying potassium channel and β-arrestin signaling pathway, resulting in decreased respiratory frequency and amplitude. Benzodiazepines enhance inhibitory neurotransmission mediated by γ-aminobutyric acid receptors, reduce the sensitivity of chemoreceptors to PaCO₂ and PaO₂, and lead to a decreased tidal volume and upper airway obstruction. Ketamine inhibits respiratory drive and respiratory muscle function by blocking N-methyl-D-aspartic acid  receptors and indirectly affecting the μ-opioid receptor. In addition, propofol inhibits pre-expiratory neuronal activity and relaxes upper airway muscles by activating the GABA_A receptor β₃ subunit. Currently, specific antagonists (naloxone/flumazenil) and respiratory stimulants (doxapram) are clinically used to treat respiratory depression, but they have defects such as short duration of action and insufficient specificity. The development of novel stimulants targeting μ-opioid receptor agonists and the D-serine release pathway of astrocytes, as well as broad-spectrum antidotes based on "molecular cage" technology, has become a new sphere of research that aims at precisely reversing respiratory depression while preserving analgesic and sedative effects. This article reviews the biological mechanisms of respiratory depression caused by sedative hypnotic anesthetic drugs, explores the advantages and disadvantages of treatments currently availabe, and proposes new strategies for improving respiratory depression in the future.