Safety and efficacy of Compound Huangdai Tablets combined with all-trans retinoic acid for treatment of acute promyelocytic leukemia: Clinical evidence and potential mechanisms / 中草药·英文版

Objective: To evaluate the safety and efficacy of Compound Huangdai Tablets (Realgar-Indigo Naturalis formula, RIF) combined with all-trans retinoic acid (ATRA) to treat acute promyelocytic leukemia (APL). Methods: This study was registered in PROSPERO (CRD42018108118). The relevant literatures on RIF treatment of APL were systematically searched in the following databases: China National Knowledge Infrastructure, Wanfang, VIP Medical Information System, Chinese Biomedical Database, EMBASE, Cochrane Library, and PubMed. The quality of the included studies was evaluated and Review Manager 5.3 software and Stata 13.0 software were used to perform the Meta-analysis. In addition, this study used the method of network pharmacology to conduct a preliminary exploration of the mechanism of RIF on APL. Results: The study included 12 studies involving 775 APL patients. The Meta-analysis showed that there was no significant difference (P 0.05) between the RIF group and the arsenic trioxide (ATO) group for primary outcomes, secondary outcomes apart from liver dysfunction. The incidence of liver dysfunction (P = 0.006) in the RIF group were significantly lower than those in the ATO group. In addition, the cost of maintenance therapy in the RIF group was significantly lower (P 0.05) than the ATO group. Besides, the active ingredients in RIF mainly act on targets proteins such as ACHE, NCOA2, RXRA, and then play a role in the treatment of APL through regulating multiple molecular mechanisms, such as TP53 regulates transcription of cell cycle genes, nuclear receptor transcription pathway. Conclusion: There was no significant difference in efficacy of oral RIF combined with ATRA compared with intravenous ATO combined with ATRA for the treatment of APL. The oral RIF exposed patients to less risk, offered more convenience and had lower prices. RIF can treat APL by multi-target and multi-pathway interventions that inducing apoptosis of APL cells and inhibiting the proliferation of APL cells, and so on. Therefore, oral RIF in the treatment of APL is worthy of further research and development.

Effect of Main Component of Realgar Arsenic Disulfide （As2S2） on DNA Methylation of SKM-1 Cells with Myelodysplastic Syndrome / 中国实验方剂学杂志

ObjectiveTo explore the effects of the main component of Realgar arsenic disulfide （As2S2） on DNA methylation of SKM-1 cells with myelodysplastic syndrome. MethodCell Counting Kit-8 （CCK-8） was used to detect the inhibitory effect of As2S2（0， 1， 2， 4， 8， 16 μmol·L-1）on SKM-1 cells. Propidium iodide （PI） staining was applied to detect the effect of As2S2（0， 1， 2， 4 μmol·L-1）on the SKM-1 cell cycle. The effect of As2S2 （0， 4 μmol·L-1） on the methylation of SKM-1 cells on a genome-wide scale was observed by using Human Methylation 850K BeadChip， followed by Kyoto Encyclopedia of Genes and Genomes （KEGG） and gene ontology （GO） analyses. According to the microarray data， the antioncogene TUSC3 was selected， and real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot were adopted to investigate the effect of As2S2 （0， 1， 2， 4 μmol·L-1） on the mRNA and protein expression of TUSC3， respectively. ResultCompared with the conditions in the blank group， As2S2 inhibited SKM-1 cells， increased the proportion of cells in the G0/G1 phase， and decreased the proportion of cells in the S phase（P<0.05）. The 850K microarray showed that 4 μmol·L-1 As2S2 could significantly induce DNA methylation in SKM-1 cells， with 12 710 differentially methylated genes involved （50% hypermethylated and 50% hypomethylated genes）. KEGG and GO analyses showed that differentially methylated genes were involved in many important biological functions and signaling pathways， including purine metabolism， natural killer cell-mediated cytotoxicity， endocytosis， chemokine signaling pathway， and nuclear ubiquitin ligase complex. In terms of downstream gene expression， Real-time PCR and Western blot showed that As2S2 increased the expression of TUSC3， as compared with the conditions in the blank group （P<0.05）. ConclusionAs2S2， the main component of Realgar， has a significant regulatory effect on the methylation of SKM-1 cells， which is presumedly achieved by increasing the expression of TUSC3.

Realgar (α-As4S4) Treats Myelodysplastic Syndromes through Reducing DNA Hypermethylation / 中国结合医学杂志

DNA hypermethylation is an epigenetic modification that plays a critical role in the oncogenesis of myelodysplastic syndromes (MDS). Aberrant DNA methylation represses the transcription of promotors of tumor suppressor genes, inducing gene silencing. Realgar (α-As4S4) is a traditional medicine used for the treatment of various diseases in the ancient time. Realgar was reported to have efficacy for acute promyelocytic leukemia (APL). It has been demonstrated that realgar could efficiently reduce DNA hypermethylation of MDS. This review discusses the mechanisms of realgar on inhibiting DNA hypermethylation of MDS, as well as the species and metabolisms of arsenic in vivo.

In vitro inhibitory effects of realgar nanoparticles on MCF- 7 breast cancer stem cells / 中国药房

OBJECTIVE To study in vitro inhibitory effects of realgar nanoparticles on breast cancer stem cells. METHODS Human breast cancer MCF- 7 parent cells were selected as subjects and cultured by serum-free culture to obtain breast cancer stem cells. Using adriamycin （1 mg/L）as positive control ，same concentration of water-processed realgar as reference ，the effects of realgar nanoparticles on the proliferation of MCF- 7 parent cells and stem cells were detected by CCK- 8 method. The effects of realgar nanoparticles on the formation of mammosphere ，the ability of differentiation ，migration and invasion ，the proportion of CD44+/CD24－ subgroup in breast cancer stem cells were detected by mammosphere formation and differentiation experiment ， scratch experiment ，Transwell invasion experiment and flow cytometry. Western blot assay was used to detect the expression of proteins related to epithelial mesenchymal transformation pathway （E-cadherin and vimentin ） in breast cancer stem cells. RESULTS The survival rates of MCF- 7 parent cells and stem cells （except for breast cancer stem cells in both 1 mg/mL groups ）in 1，5，10，40，60，80 mg/L groups of water-processed realgar and realgar nanoparticles were significantly lower than blank control group（P＜0.01）. The number of mammosphere （＞20 stem cells ）in 1，2.5，5，10 mg/L groups of water-processed realgar and realgar nanoparticles was significantly lower than blank control group （P＜0.01）；the volume of mammosphere decreased and the differentiated adherent cells decreased ；the healing rate of wound ，relative invasion rate （except for water-processed realgar 1 mg/L group）and the proportion of CD 44+/CD24－ subgroup were significantly lower than blank control group （P＜0.01）. The expressions of E-cadherin in 2.5，10 mg/L groups of water-processed realgar and realgar nanoparticles was significantly higher than blank control group ，and the expressions of vimentin was significantly lower than those in blank control group （P＜0.01）. The above effects of realgar nanoparticles were generally better than those of water-processed realgar with the same mass concentration （P＜ 0.01）. CONCLUSIONS Compared with water-processed realgar with the same mass concentration ，realgar nanoparti cles can significantly inhibit the proliferation of breast cancer stem cells， the formulation and differential ability of mammo- sphere，and reduce the proportion of CD 44+/CD24－ subgroup. The effect may be associated with the inhibition of migration and invasion of breast cancer stem cells by inhibiting the expression of proteins related to epithelial mesenchymal transformation pathway.

Effect of Arsenic in Realgar on Rat Ability in Novel Objects Recognition Test / 中国实验方剂学杂志

Objective:To determine the contents of inorganic arsenic（iAs），monomethylarsonic acid（MMA） and dimethylarsinic acid（DMA） in brain tissues and blood by using hydride generation-cold trap-atomic absorptionspectrometry（HG-CT-AAS）， and to explore the toxic effects of Realgar on central nervous system of rats. Method:The 96 Wistar rats were randomly divided into 4 groups：normal control group，0.3，0.9 and 2.7 g·kg^-1 Realgar groups. They then received intragastric administration for 14，28 and 42 days respectively， so a total of 12 groups were formed， with 8 animals in each group. The normal group was given the same dose of sodium carboxymethyl cellulose （CMC-Na） by gavage. The contents of iAs，MMA and DMA in blood and brain tissues were determined by HG-CT-AAS. The novel object recognition test was conducted to observe the learning and memory ability of rats. The changes of hippocampal neuron ultrastructure were observed by transmission electron microscopy. Result:There was no difference in the growth，weight and hippocampal coefficient of the experimental animals. The method of HG-CT-AAS showed a good linearity，precision，accuracy and recovery in content determination of arsenic （at various forms） in rat brain and blood. MMA and DMA were detected in the brain of realgar groups at time-dose-effect relationship. iAs，MMA and DMA were detected in the blood of Realgar groups. The nuclear membrane， mitochondria and endoplasmic reticulum in hippocampus neurons of rats were gradually damaged with the increase of Rhubarb exposure dose and time. After 14 days of exposure to Realgar，compared with the normal control group，there was no significant difference in the novel object recognition index among Realgar groups. After 28 days of exposure，only 2.7 g·kg^-1 Realgar group showed statistically significant difference with the control group （<italic>P</italic><0.05）. After 42 days of exposure， the novel object recognition index of 0.9 and 2.7 g·kg^-1 Realgar groups was significantly lower than that in normal control group（<italic>P</italic><0.05）. Conclusion:The metabolites of Realgar in rats are iAs，MMA and DMA. MMA and DMA can be accumulated in the brain tissue through the blood-brain barrier，causing the decline of the ability of learning and memory and leading to damage of hippocampal neurons.

Arsenic-Containing Qinghuang Powder () is an Alternative Treatment for Elderly Acute Myeloid Leukemia Patients Refusing Low-Intensity Chemotherapy / 中国结合医学杂志

OBJECTIVE@#To analyze the overall survival (OS) of elderly acute myeloid leukemia (AML) patients treated with oral arsenic-containing Qinghuang Powder (, QHP) or low-intensity chemotherapy (LIC).@*METHODS@#Forty-two elderly AML patients treated with intravenous or subcutaneous LIC (1 month for each course, at least 3 courses) or oral QHP (3 months for each course, at least 2 courses) were retrospectively analyzed from January 2015 to December 2017. The main endpoints of analysis were OS and 1-, 2-, 3-year OS rates of patients, respectively. And the adverse reactions induding bone marrow suppression, digestive tract discomfort and myocardia injury were observed.@*RESULTS@#Out of 42 elderly AML patients, 22 received LIC treatment and 20 received QHP treatment, according to patients' preference. There was no significant difference on OS between LIC and QHP patients (13.0 months vs. 13.5 months, >0.05). There was no significant difference on OS rates between LIC and QHP groups at 1 year (59.1% vs. 70.0%), 2 years (13.6% vs. 15%), and 3 years (4.6% vs. 5.0%, all >0.05). Furthermore, there was no significant difference of OS on prognosis stratification of performance status > 2 (12 months vs. 12 months), age> 75 year-old (12.0 months vs. 12.5 months), hematopoietic stem cell transplant comorbidity index >2 (12 months vs. 13 months), poor cytogenetics (12 months vs. 8 months), and diagnosis of secondary AML (10 months vs. 14 months) between LIC and QHP patients (>0.05).@*CONCLUSION@#QHP may be an alternative treatment for elderly AML patients refusing LIC therapy.

Effect of Metabolic Reprogramming on Toxin Eliminating Therapeutics in Targeting Lung Cancer Stem Cells / 中国实验方剂学杂志

Objective::To observe the effect of realgar nanoparticles (a representative drug in toxin eliminating therapeutics) targeting hypoxia-inducible factors (HIF), which act as effector molecules on metabolic reprogramming of lung cancer stem cells, and to explore the effect mechanism of lung cancer stem cells and metabolic reprogramming in the process of lung cancer metastasis, so as to verify the effectiveness of toxin eliminating therapeutics in the prevention and treatment of lung cancer metastasis. Method::Lung cancer A549 cells were cultured in vitro, and lung cancer stem cells were then identified and selected. The stem cells were divided into blank control group, cisplatin group (5 mg·L-1), realgar nanoparticles low, medium and high dose groups (100, 200, 400 mg·L-1). After intervention, glucose oxidase method was used to detect the effect of realgar nanoparticles on the glucose metabolism of lung cancer stem cells, real-time polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression levels of hypoxia-inducible factors-1α (HIF-1α), C-myc and p53, while Western blot was used to detect the expression of related proteins HIF-1α, phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and mammalian target of rapamycin (mTOR), and enzyme linked immunosorbent assay (ELISA) was used to detect the glucose transporter 1 (GLUT1), pyruvate dehydrogenase kinase 1 (PDK1), pyruvate kinase M (PKM), phosphofructokinase(PFK), pyruvate dehydrogenase (PDH) and lactic dehydrogenase (LDH) expression. Result::As compared with the blank control group, realgar nanoparticles can reduce the glucose consumption of lung cancer stem cells, and the glucose consumption was reduced with the increase of dose in a time-and dose-dependent manner (P<0.01). Realgar nanoparticles can inhibit the mRNA expression of HIF-1α, a key factor in metabolic reprogramming of lung cancer stem cells (P<0.05, P<0.01), down-regulated C-myc mRNA and up-regulated the p53 mRNA expression (P<0.05, P<0.01), down-regulated protein expressions of PI3K, Akt, mTOR(P<0.05, P<0.01), and inhibited the expression of related enzymes GLUT1, PDK1, PFK, PKM, PDH, and LDH levels (P<0.05, P<0.01). With the increase of dose, the regulation and control ability of realgar nanoparticles gradually increased. Conclusion::Toxin eliminating therapeutics can drive the metabolic reprogramming of lung cancer stem cells by targeting HIF effector molecule, and then inhibit the invasion and metastasis of lung cancer.

Mechanism of Liushenwan and Realgar in Inducing Apoptosis and DNA Damage in Human Endometrial Cancer JEC Cells / 中国实验方剂学杂志

Objective:Compare the anti-tumor effect and mechanism of Liushenwan and realgar (As4S4) on human endometrial cancer cells JEC. Method:The release of As in Liushenwan and As4S4 was measured by atomic absorption spectrometry. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used for cell proliferation, and cell migration was measured by Transwell assay. Flow cytometry and Western Blot were used to determine apoptosis and DNA damage. Result:The dissolution of As in Liushenwan was 17.4%, and that of As4S4 was only 1.6% according to atomic absorption assay. With the same content of As, compared with the As4S4 group, the cell viability in the 3，10 mg·L-1 Liushenwan groups was decreased (P<0.05), the early apoptosis rate was significantly increased in 0.25，0.5，1 mg·L-1 Liushenwan groups (P<0.05), the rate of cell migration was decreased in 1 mg·L-1 Liushenwan group (P<0.05), the expressions of cleaved cysteinyl aspartate-specific proteinase-3 (cleaved Caspase-3), cleaved cysteinyl aspartate-specific proteinase-7 (cleaved Caspase-7), cleaved poly ADP-ribose polymerase (cleaved PARP), phosphorylated histone (p-H2AX), phosphorylation of checkpoint kinase 2 (p-CHK2) and ataxia-telangiectasia mutated (ATM) were increased in 1 mg·L-1 Liushenwan group (P<0.05), while the expression of phosphorylation of ataxia-telangiectasia mutated rad3-related (ATR) was decreased in 1 mg·L-1 Liushenwan group (P<0.05), with no significant changes in the expressions of cysteinyl aspartate-specific proteinase-3 (Caspase-3) and cysteinyl aspartate-specific proteinase-7 (Caspase-7). Conclusion:With the same content of As, both Liushenwan and As4S4 could inhibit JEC cell proliferation and migration, and induce cell apoptosis and DNA damage. Liushenwan has a stronger effect than As4S4. It is suggested that there are other components in Liushenwan with an anti-tumor effect in cooperation with As.

Analysis of Effect of Realgar with Different Compatibility Proportions on Transitional Constituents of Indigo Naturalis in Rat Serum Based on UPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective:To investigate the effect of different dose of Realgar compatible with Indigo Naturalis on the transitional constituents of Indigo Naturalis in rat serum based on the compatibility of Qinghuangsan.Method:Indigo Naturalis test solution, the drug-containing serum of three different proportions of Qinghuangsan (10 g of Indigo Naturalis compatible with 52.5, 105, 210 mg of Realgar for group A, B and C, respectively) and blank serum were detected by UPLC-Q-TOF-MS/MS, in combination with the chemical components identified in Indigo Naturalis test solution, the differences of transitional constituents of Indigo Naturalis in rat serum from the group A, B and C were analyzed. HL-60 cells (human leukemia cells) were treated with the three groups of Qinghuangsan drug-containing serum and the effect of drug-containing serum on the activity of HL-60 cells was detected by cell counting kit-8 (CCK-8) assay.Result:A total of 19, 22, 25 of transitional constituents were detected in Qinghuangsan drug-containing serum from group A, B and C, respectively. The three groups of drug-containing serum all contained 5 prototype components from Indigo Naturalis test solution, including tryptanthrin, indigo, indirubin, 2-aminobenzoic acid and N-phenyl-2-naphthylamine, respectively. The results of CCK-8 assay showed that Qinghuangsan drug-containing serum of group C had the strongest inhibitory effect on HL-60 cells.Conclusion:After fixed Indigo Naturalis dose, with the increase of Realgar dose, the transitional constituents in rat serum increase and the inhibitory effect on HL-60 cells also gradually enhances, which indicates that Realgar may promote the absorption of active components in Indigo Naturali in vivo, thus enhance the efficacy, further explains the compatibility law and pharmacodynamic material basis of different proportions of Realgar and Indigo Naturalis.

Research of Anticancer Realgar Preparations from Perspective of Patent / 中国实验方剂学杂志

To review the research progress of anticancer realgar preparations from the perspective of patent，in order to provide a reference for the research of new antitumor realgar drugs. IncoPat database was retrieved with keywords plus IPC classification number. Original data and 72 patents of anti-cancer realgar preparations were obtained and included after artificial denoising. The quantitative analysis was made on the information about application trends，application types and patentee. The technical points of representative patents were summarized. ① The patent types of anticancer realgar preparations are mainly product patents. Its technological innovation has undergone the development of realgar compound agents，arsenic sulfide single agents，nanometer products of realgar agents，bioleaching of realgar preparations，compound realgar extract preparations，compound nano realgar preparations and arsenic sulfide polymorphic crystalline structure. ② Nano realgar agents， realgar bioleaching preparations， new crystalline type realgar preparations and realgar compound preparations. ③ The following studies should be strengthened in the future，involving the comparison of biological effects and toxicity of nano realgar preparations of different preparation processes，the selection of optimal particle size，the druggability of realgar bioleaching preparations，the discovery of new As4S4 crystalline forms and the secondary development of anticancer realgar preparations.

Bioleaching of realgar nanoparticles using the extremophilic bacterium Acidithiobacillus ferrooxidans DLC-5

BACKGROUND: This paper presents micro- and nano-fabrication techniques for leachable realgar using the extremophilic bacterium Acidithiobacillus ferrooxidans (A. ferrooxidans) DLC-5. RESULTS: Realgar nanoparticles of size ranging from 120 nm to 200 nm were successfully prepared using the highenergy ball mill instrument. A. ferrooxidans DLC-5 was then used to bioleach the particles. The arsenic concentration in the bioleaching system was found to be increased significantly when compared with that in the sterile control. Furthermore, in the comparison with the bioleaching of raw realgar, nanoparticles could achieve the same effect with only one fifth of the consumption. CONCLUSION: Emphasis was placed on improving the dissolvability of arsenic because of the great potential of leachable realgar drug delivery in both laboratory and industrial settings

Improvement of glycyrrhetinic acid on damage of synaptic ultrastructures of hippocampus induced by realgar in mice / 吉林大学学报(医学版)

Objective: To explore the improvement of glycyrrhetinic acid (G A) on the damage of synaptic ultrastructures of hippocampus induced by realgar in the mice, and to clarify the related mechanisms. Methods: Sixty ICR mice were randomly divided into three groups with twenty mice in each group: control group (intragastrically treated with 0. 5 % CMC-Na), realgar group (intragatrically treated with realgar 1. 35 g • kg- 1), and GA intervention group (intragastrically administered with GA 48 mg • kgx and realgar 1. 35 g • kg1). The mice were administrated once a day for eight consecutive weeks. The cognitive and memory abilities were tested using object recognition task (ORT). The levels of glutathione (GSH) in the hippocampus in the mice in various groups were detected. The changes of the ultrastructures of synapse in hippocampal CA1 region, the width of synaptic cleft, the length of synaptic active zone, the thickness of post synaptic density (PSD) and the curvature of synaptic interface were observed by transmission electron microscope. Results: Compared with control group, the preferential index (PI) for the novel object and the level of GSH in the hippocampus tissue of the mice in realgar group were significantly decreased (P 0 . 05), and the level of GSH in the hippocampus tissue of the mice in GA intervention group was significantly increased (P 0 . 05). Conclusion: Realgar can change the synaptic structural parameters and cause deficits in cognitive and memory abilities. GA can alleviate the abnormal ultrastructural changes in the hippocampal synapses of the mice.

Modern Pharmacological Toxicology and Clinical Application of Antitumor Nanometer Realgar / 中国实验方剂学杂志

Realgar is toxic and belongs to drug of poison attack, with anti-cancer, anti-pest, dry wet and expectorant effects. Ancient doctors often used realgar to treat carbuncle, boil, abdominal pain and other diseases. Modern doctors use it to treat malignant tumors and blood diseases. The toxicity of realgar results in a small range of safety in its drug use. Modern scholars combine traditional Chinese medicine with nano-technology means to grind realgar into nano-realgar. Compared with realgar, nano-realgar has an improved bioavailability and reduced toxicity in vivo. Modern pharmacological researches use nano-realgar to interven lung cancer cells, skin cancer cells, cervical cancer cells, ovarian cancer cells and leukemia cells in experiments, which confirmed that nano-realgar has effects in inhibiting cell proliferation, inducing apoptosis and cell differentiation, inhibiting nucleic acid synthesis and angiogenesis, but with antiviral, sterilization of analgesic effects. Modern toxicological studies have been conducted in mice through intragastric treatment with different concentrations of nano-realgar preparation. The symptoms and signs of mice and various auxiliary examination indexes were recorded at different time periods. It was concluded that nano-realgar has regular effects on blood biochemical indexes at safe doses. In clinical trials, nano-realgar was applied to treat damaged wound surface of malignant tumor. It was found that nano-realgar can promote healing of wound surface of tumor, alleviate pain, relieve clinical symptoms, such as bleeding, purulence and odor, and improve quality of life of patients. In addition, with a simple usage and good patient compliance, administration with nano-realgar requires no hospitalization, can save medical resources, and is worthy of clinical promotion and application.

Research progress in mineral Chinese medicine realgar / 中国中药杂志

Realgar is a mineral traditional medicine with definite efficacy. The function of realgar is detoxicating, insecticiding, eliminating dampness and phlegm, etc. It is widely applied in clinical practice by compatibility medicines. However, the safety and scientificalness of clinical application are questioned because of the toxic effect caused by arsenic compounds. At present, there are still many problems in the research of realgar, which are mainly manifested in three areas: the expression of main components and effective substances are inconsistent; the anti-tumor mechanism is difficult to explain at the molecular level; the mechanism of compatibility is not clear. As a result, realgar and realgar-containing Chinese patent medicines are frequently prohibited from entering the international market, and the reputation of traditional Chinese medicine is also damaged. This paper would analyze the research status of realgar at home and abroad as well as its problems from its main components, effective substances, anti-tumor mechanism and compatibility mechanism. In view of these difficulties, quantum chemical calculation method is proposed to solve them, so as to make up for the shortcomings and limitations of experimental technology and experimental conditions, reduce the cost of realgar research and improve research efficiency. Moreover, it provides inspiration for research of other mineral medicine.

Association of Gene Mutations with Response to Arsenic-Containing Compound Qinghuang Powder () in Patients with Myelodysplastic Syndromes / 中国结合医学杂志

OBJECTIVES@#To investigate the relationship between gene mutations and response to Compound Qinghuang Powder (, CQHP) in patients with myelodysplastic syndrome (MDS).@*METHODS@#Forty-three MDS patients were genotyped by ultra-deep targeted sequencing and the clinical data of patients were collected and the relationship between them was analyzed.@*RESULTS@#Up to 41.86% of patients harbored genet mutations, in most cases with more than one mutation. The most common mutations were in SF3B1, U2AF1, ASXL1, and DNMT3A. After treatment with CQHP, about 88.00% of patients no longer required blood transfusion, or needed half of prior transfusions.@*CONCLUSIONS@#CQHP is an effective treatment for patients with MDS, especially those with gene mutations in SF3B1, DNMT3A, U2AF1, and/or ASXL1.

Relation of Blood Arsenic Concentration with Effect and Safety of Arsenic-Containing Qinghuang Powder () in Patients with Myelodysplastic Syndrome / 中国结合医学杂志

OBJECTIVE@#To investigate the relation of blood arsenic concentration (BAC) with clinical effect and safety of arsenic-containing Qinghuang Powder (, QHP) in patients with myelodysplastic syndrome (MDS).@*METHODS@#Totally 163 patients with MDS were orally treated with QHP for 2 courses of treatment, 3 months as 1 course. The BACs of patients were detected by atomic fluorescence spectrophotometry at 1, 3, and 6 months during the treatment, and the effective rate, hematological improvement and safety in patients after treatment with QHP were analyzed.@*RESULTS@#After 2 courses of treatment, the total effective rate was 89.6% (146/163), with 31.3% (51/163) of hematological improvement and 58.3% (95/163) of stable disease. The hemoglobin increased from 73.48 ± 19.30 g/L to 80.39 ± 26.56 g/L (P0.05). Among 46 patients previously depended on blood transfusion, 28.3% (13/46) completely got rid of blood transfusion and 21.7% (10/46) reduced the volume of blood transfusion by more than 50% after treatment. The BACs were significantly increased in patients treated for 1 month with 32.17 ± 18.04 μ g/L (P0.05). The adverse reactions of digestive tract during the treatment were mild abdominal pain and diarrhea in 14 cases (8.6%), and no patients discontinued the treatment. The BACs of patients with gastrointestinal adverse reactions were significantly lower than those without gastrointestinal adverse reactions (22.39 ± 10.38 vs. 37.89 ± 11.84, μ g/L, P<0.05). The BACs of patients with clinical effect were significantly higher than those failed to treatment (40.41 ± 11.69 vs. 23.84 ± 12.03, μ g/L, P<0.05).@*CONCLUSION@#QHP was effective and safe in the treatment of patients with MDS and the effect was associated with BACs of patients.

Determination of arsenic species in serum, liver, kidney, and spleen of realgar nanoparticles treated rats by HPLC-ICP-MS / 中草药

Objective Quantitative analysis of four arsenic species As (III), As (V), monomethyl arsenate (MMA), dimethyl arsenate (DMA) in rat serum, liver, kidney, and spleen was performed to compare their differences between realgar and realgar nanoparticles (NPs) groups. Methods SD rats were ig treated with blank solvent, realgar, and realgar NPs (800 mg/kg) respectively. After 28 d of continuous administration, serum and tissues were collected and four arsenic species were determined by high performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS). Results Four arsenic species were detected in serum and kidney of rats, three were detected in the liver and two in the spleen. The content of arsenic species in the realgar NPs group was significantly higher than that in the realgar group. Conclusion Nanotechnology enhanced the bioavailability of realgar, and more arsenic was absorbed into the body and underwent metabolic transformation, which might lead to increased toxicity of realgar NPs.

Quality Control of Realgar in Compound Huanglian Ointment / 中国中医药信息杂志

Objective To control the quality of realgar in compound Huanglian Ointment.Methods As2O3which is the toxic component of realgar was carried limit test by Gutzeit's test; Potassium sulfate, ammonium sulfate and sulfuric acid were used to digest, and then titration method was used to determine the content of As2S2in compound Huanglian Ointment. Results The content of the soluble As in compound Huanglian Ointment was no higher than 15.6 μg/g. The content of As2S2in compound Huanglian Ointment was no less than 2.21 mg/g. Conclusion The method is simple and easy,which can be applied as the quantity control method of compound Huanglian Ointment.

Synergistic effect and different toxicities of adjuvant components of Realgar–Indigo Naturalis formula / 中草药·英文版

Objective: Realgar–Indigo Naturalis formula (RIF) is a well-known arsenic-containing preparation that is used to treat acute promyelocytic leukemia (APL) in China. In recent multicenter clinical trials, complete remission rates in APL patients have ranged from 96.08% to 100%. RIF has a satisfactory therapeutic effect, but its safety is a widespread concern, since the preparation contains arsenic, a wide-ranging and naturally occurring toxicant. In this study, in order to determine the toxic potential of RIF, acute toxicity and sub chronic toxicity assays were performed to evaluate the toxic potential of Realgar and the adjuvant components of RIF in addition to Realgar's synergy with these adjuvant components. Methods: KM mice and Wistar rats were selected for these experiments. To evaluate acute toxicity, the toxic effects of a single dose of a gradient of concentrations of Realgar were firstly determined. Then, the toxic effects of combinations of gradient doses of Realgar and fixed doses of Indigo naturalis and Salvia miltiorrhiza were evaluated. Results: The results showed that when Realgar was used alone, the LD50 was 2756.73 mg/kg (equivalent to 23.6 mg/kg As2O3). However, the LD50 dropped to 936.90 mg/kg when Realgarwas used with I. naturalis. By contrast, the LD50 increased to 7538.86 mg/kg when Realgar was used with S. miltiorrhiza. Hence, I. naturalis strengthened the toxicity of Realgar, whereas S. miltiorrhiza displayed the opposite effect. The sub chronic toxicity assessment results revealed a trend that was consistent with acute toxicity. Changes in the levels of different valence states of arsenic were also taken into account. The test results of the effects of in vitro combinations of Realgar and adjuvant components on soluble arsenic dissolution showed that I. naturalis increased the level of soluble arsenic in Realgar extracts and I. naturalis suspensions when theRealgar/I. naturalis ratio was 2, 1.5, and 1.0. However, S. miltiorrhiza did not affect it. Conclusion: Based on the collective experimental results presented here, it can be concluded that the toxicity of RIF is the result of the soluble arsenic in Realgar and that the I. naturalis and S. miltiorrhiza in the RIF exert completely opposite effects on the toxicity of Realgar. This maybe an intelligent explanation for the compatibility of this formula, and this RIF study may therefore be viewed as a classic case of traditional Chinese medicine research on compatibility.

Arsenic tolerance and bioleaching from realgar based on response surface methodology by Acidithiobacillus ferrooxidans isolated from Wudalianchi volcanic lake, northeast China

Background: Traditional methods of obtaining arsenic have disadvantages such as high cost and high energy consumption. Realgar is one of the most abundant arsenic sulphide minerals and usually treated as waste in industry. The aim of the present study was to screen an arsenic tolerant bacterium used for bioleaching arsenic from realgar. Results: An acidophilic iron-oxidizing bacterium BYQ-12 was isolated from Wudalianchi volcanic lake in northeast China. BYQ-12 was a motile, rod-shaped gram-negative bacterium with an optimum growth at 30°C and pH 2.5. 16S rDNA phylogeny showed that BYQ-12 was a new strain of Acidithiobacillus ferrooxidans. The inhibitory concentrations (ICs) of arsenite and arsenate were 32 and 64 mM, respectively. A significant second-order model was established using a Box­Behnken design of response surface methodology (BBD-RSM) and it estimated that a maximum arsenic bioleaching rate (73.97%) could be obtained when the pulp concentration, pH and initial ferrous ion concentration were set at optimized values of 0.95% w/v, 1.74 and 3.68 g/L, respectively. SEM, EDS and XRD analyses also revealed that there was direct bioleaching besides indirect electrochemical leaching in the arsenic bioleaching system. Conclusion: From this work we were successful in isolating an acidophilic, arsenic tolerant ferrous iron-oxidizing bacterium. The BBD-RSM analysis showed that maximum arsenic bioleaching rate obtained under optimum conditions, and the most effective factor for arsenic leaching was initial ferrous ion concentration. These revealed that BYQ-12 could be used for bioleaching of arsenic from arsenical minerals.