1.The toxicity of ZnO and CuO nanoparticles on biological wastewater treatment and its detoxification: a review.
Yuran YANG ; Can ZHANG ; Zhenlun LI
Chinese Journal of Biotechnology 2023;39(3):1026-1039
		                        		
		                        			
		                        			The wide use of ZnO and CuO nanoparticles in research, medicine, industry, and other fields has raised concerns about their biosafety. It is therefore unavoidable to be discharged into the sewage treatment system. Due to the unique physical and chemical properties of ZnO NPs and CuO NPs, it may be toxic to the members of the microbial community and their growth and metabolism, which in turn affects the stable operation of sewage nitrogen removal. This study summarizes the toxicity mechanism of two typical metal oxide nanoparticles (ZnO NPs and CuO NPs) to nitrogen removal microorganisms in sewage treatment systems. Furthermore, the factors affecting the cytotoxicity of metal oxide nanoparticles (MONPs) are summarized. This review aims to provide a theoretical basis and support for the future mitigating and emergent treatment of the adverse effects of nanoparticles on sewage treatment systems.
		                        		
		                        		
		                        		
		                        			Wastewater/toxicity*
		                        			;
		                        		
		                        			Sewage/chemistry*
		                        			;
		                        		
		                        			Zinc Oxide/chemistry*
		                        			;
		                        		
		                        			Waste Disposal, Fluid
		                        			;
		                        		
		                        			Nanoparticles/chemistry*
		                        			;
		                        		
		                        			Metal Nanoparticles/chemistry*
		                        			;
		                        		
		                        			Nitrogen/metabolism*
		                        			;
		                        		
		                        			Water Purification
		                        			
		                        		
		                        	
2.Quality evaluation of Galli Gigerii Endothelium Corneum based on HPLC fingerprints and content determination of nucleosides.
Jia FAN ; Xiao-Qian LIU ; Chen-Xiao-Ning MENG ; Sen JIAO ; Wei-Hong FENG ; Li-Hua YAN ; Zhi-Min WANG
China Journal of Chinese Materia Medica 2023;48(1):114-125
		                        		
		                        			
		                        			Galli Gigerii Endothelium Corneum(GGEC), the dried gizzard membrane of Gallus gallus domesticus is a Chinese medicinal material commonly used for digestion. However, due to the particularity of texture and composition, its active ingre-dients have not been clarified so far, and there is also a lack of quality evaluation indicators. In this study, UPLC-Q-TOF-MS was used to analyze the chemical components from the water extract of GGEC, and ten nucleosides were identified for the first time. HPLC fingerprints of the water extracts of GGEC were established and the content of seven nucleosides was determined. The fingerprint similarities of 40 batches of GGEC samples ranged from 0.765 to 0.959, indicating that there were great differences among the GGEC products processed with different methods. In addition, SPSS 22.0 and SIMCA 14.1 were used for hierarchical cluster analysis(HCA) and principal component analysis(PCA) on the 19 common peaks of the HPLC fingerprints of GGEC, and the 40 batches of samples were divided into three categories: raw GGEC, fried GGEC and vinegar-processed GGEC. Eight differential components in GGEC were marked by orthogonal partial least squares discrimination analysis(OPLS-DA), two of which were adenine and thymine. The results of content determination showed that the total content of the seven nucleosides in raw GGEC, fried GGEC and vinegar-processed GGEC were 182.5-416.8, 205.3-368.7, and 194.2-283.0 μg·g~(-1), respectively. There were significant differences in the content of hypoxanthine, thymine and thymidine among the GGEC products processed with different methods(P<0.05), which were graded in the order of fried GGEC>vinegar-processed GGEC>raw GGEC. This suggested that the content of hypoxanthine, thymine and thymidine tended to increase during the frying process, and the variation range might be related to the degree of heat exposure. The established methods in this study were simple and reproducible, and could be used for qualitative and quantitative analysis of GGEC and its processed pro-ducts. This study also provided reference for the establishment of quality standards of GGEC with chemical components as control index.
		                        		
		                        		
		                        		
		                        			Nucleosides
		                        			;
		                        		
		                        			Drugs, Chinese Herbal/chemistry*
		                        			;
		                        		
		                        			Chromatography, High Pressure Liquid
		                        			;
		                        		
		                        			Acetic Acid
		                        			;
		                        		
		                        			Thymine
		                        			;
		                        		
		                        			Thymidine
		                        			;
		                        		
		                        			Water
		                        			;
		                        		
		                        			Hypoxanthines
		                        			
		                        		
		                        	
3.Establishment of a quantitative method for GC analysis of polyoxyethylene (35) castor oil in microemulsion extracts.
Yan-Jing WANG ; Yi CHENG ; Ze-Min OU ; Yao ZHANG ; Lin YAN ; Yan TONG ; Jin-Yu WANG ; De-Wen LIU
China Journal of Chinese Materia Medica 2023;48(22):6075-6081
		                        		
		                        			
		                        			With the continuous exploration of microemulsions as solvents for traditional Chinese medicine extraction, polyoxyethy-lene(35) castor oil(CrEL), a commonly used surfactant, is being utilized by researchers. However, the problem of detecting residues of this surfactant in microemulsion extracts has greatly hampered the further development of microemulsion solvents. Based on the chemical structures of the components in CrEL and the content determination method of castor oil in the 2020 edition of the Chinese Pharmacopoeia(Vol. Ⅳ), this study employed gas chromatography(GC) and single-factor experiments to optimize the preparation method of methyl ricinoleate from CrEL. The conversion coefficient between the two was validated, and the optimal sample preparation method was used to process microemulsion extracts of Zexie Decoction from three batches. The content of methyl ricinoleate generated was determined, and the content of CrEL in the microemulsion extracts of Zexie Decoction was calculated using the above conversion coefficient. The results showed that the optimal preparation method for CrEL was determined. Specifically, 10 mL of 1 mol·L~(-1) KOH-methanol solution was heated at 60 ℃ for 15 min in a water bath. Subsequently, 10 mL of boron trifluoride etherate-methanol(1∶3) solution was heated at 60 ℃ for 15 min in a water bath, followed by extraction with n-hexane twice. CrEL could stably produce 20.84% methyl ricinoleate. According to this conversion coefficient, the average mass concentration of CrEL in the three batches of Zexie Decoction microemulsion extracts was 11.94 mg·mL~(-1), which was not significantly different from the CrEL mass concentration of 11.57 mg·mL~(-1) during microemulsion formulation, indicating that the established content determination method of this study was highly accurate, sensitive, and repeatable. It can be used for subsequent research on microemulsion extracts of Zexie Decoction and provide a reference for quality control of other drug formulations containing CrEL.
		                        		
		                        		
		                        		
		                        			Polyethylene Glycols/chemistry*
		                        			;
		                        		
		                        			Castor Oil
		                        			;
		                        		
		                        			Methanol
		                        			;
		                        		
		                        			Surface-Active Agents/chemistry*
		                        			;
		                        		
		                        			Solvents
		                        			;
		                        		
		                        			Water/chemistry*
		                        			;
		                        		
		                        			Emulsions/chemistry*
		                        			
		                        		
		                        	
4.Mechanism of Danggui Sini Decoction in improving kidney injury caused by blood stasis syndrome based on metabolomics and network pharmacology.
Lin-Lin FENG ; Si-Qi TANG ; Yun-Yuan NONG ; Ying HE ; Qian-Yi WANG ; Jing-Hua QIN ; Yue GUO ; Zhi-Heng SU
China Journal of Chinese Materia Medica 2023;48(24):6730-6739
		                        		
		                        			
		                        			This article analyzed the mechanism of Danggui Sini Decoction(DSD) in improving kidney injury caused by blood stasis syndrome(BSS) in rats. Firstly, 32 female SD rats were randomly divided into the following four groups: a normal group and a BSS group, both receiving an equal amount of distilled water by gavage; a normal+DSD group and a BSS+DSD group, both receiving 5.103 g·kg~(-1) DSD orally for a total of 14 days. Daily cold water bath was given to establish the BSS model, and on the 14th day, BSS rats were subcutaneously injected with 0.8 mg·kg~(-1) adrenaline. Normal rats were subjected to the water bath at 37 ℃ and injected with an equal volume of distilled water. After the experiment, 24-hour urine, serum, and kidney samples were collected for metabolomic analysis, biochemical measurements, and hematoxylin-eosin(HE) staining. The study then employed ~1H-NMR metabolomic technology to reveal the metabolic network regulated by DSD in improving BSS-induced kidney injury and used network pharmacology to preliminarily elucidate the key targets of the effectiveness of DSD. Pathological and biochemical analysis showed that DSD intervention significantly reduced inflammation and abnormal levels of blood creatinine, blood urea nitrogen, and urine protein in the kidneys. Metabolomic analysis indicated that DSD attenuated BSS-induced kidney injury primarily by regulating 10 differential metabolites and three major metabolic pathways(taurine and hypotaurine metabolism, citrate cycle, and acetaldehyde and dicarboxylic acid metabolism). Network pharmacology analysis suggested that the protective effect of DSD against BSS-induced kidney injury might be related to two key genes, ATP citrate lyase(ACLY) and nitric oxide synthase 2(NOS2), and two main metabolic pathways, i.e., arginine biosynthesis, and arginine and proline metabolism. This study, from the perspective of network regulation, provides initial insights and evidence into the mechanism of DSD in improving kidney injury induced by BSS, offering a basis for further investigation into the molecular mechanisms underlying its efficacy.
		                        		
		                        		
		                        		
		                        			Rats
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Rats, Sprague-Dawley
		                        			;
		                        		
		                        			Network Pharmacology
		                        			;
		                        		
		                        			Drugs, Chinese Herbal/chemistry*
		                        			;
		                        		
		                        			Metabolomics
		                        			;
		                        		
		                        			Kidney
		                        			;
		                        		
		                        			Arginine
		                        			;
		                        		
		                        			Water
		                        			
		                        		
		                        	
5.Recent advances in the structure and function of microbial community in anaerobic granular sludge.
Changjie GUO ; Weigang WANG ; Yayi WANG
Chinese Journal of Biotechnology 2023;39(11):4517-4533
		                        		
		                        			
		                        			Anaerobic granular sludge (AnGS), a self-immobilized aggregate containing various functional microorganisms, is considered as a promising green process for wastewater treatment. AnGS has the advantages of high volume loading rate, simple process and low excess sludge generation, thus shows great technological and economical potentials. This review systematically summarizes the recent advances of the microbial community structure and function of anaerobic granular sludge, and discusses the factors affecting the formation and stability of anaerobic granular sludge from the perspective of microbiology. Moreover, future research directions of AnGS are prospected. This review is expected to facilitate the research and engineering application of AnGS.
		                        		
		                        		
		                        		
		                        			Sewage/chemistry*
		                        			;
		                        		
		                        			Waste Disposal, Fluid
		                        			;
		                        		
		                        			Anaerobiosis
		                        			;
		                        		
		                        			Microbiota
		                        			;
		                        		
		                        			Water Purification
		                        			;
		                        		
		                        			Bioreactors/microbiology*
		                        			
		                        		
		                        	
6.Discovery, isolation and structural identification of alkaloid glycosides in six traditional Chinese medicine such as Coptis chinensis.
Ru WANG ; Liang-Jun GUAN ; Liang-Mian CHEN ; Rui PENG ; Jing-Jing ZHU ; Xiao-Qian LIU ; Hui-Min GAO ; Zhi-Min WANG
China Journal of Chinese Materia Medica 2023;48(17):4598-4609
		                        		
		                        			
		                        			Alkaloids are important active ingredients occurring in many traditional Chinese medicines, and alkaloid glycosides are one of their existence forms. The introduction of saccharide units improves the water solubility of alkaloid glycosides thus presenting better biological activity.Because of the low content in plants, alkaloid glycosides have been not comprehensively studied. In this study, ultrahigh performance liquid chromatography-quadrupole time of flight-tandem mass spectrometry(UPLC-QTOF-MS/MS) was employed to identify and analyze the alkaloid glycosides in Coptis chinensis, Phellodendron chinense, Menispermum dauricum, Sinomenium acutum, Tinospora sagittata and Stephania tetrandra. The results showed that except Tinospora sagittata, the other five herbal medicines contained alkaloid glycosides. Furthermore, the alkaloid glycosides in each herbal medicine were identified based on UV absorption spectra, quasimolecular ion peaks in MS, fragment ions information in the MS/MS, and previous literature reports. A total of 42 alkaloid glycosides were identified. More alkaloid glycosides were identified in C. chinensis and Menispermum dauricum, and eleven in C. chinensis were potential new compounds. Furthermore, the alkaloid glycosides in the water extract of C. chinensis were coarsely se-parated by macroporous adsorption resin, purified by column chromatography with D151 cation exchange resin, ODS and MCI, combined with semi-preparative high performance liquid chromatography. Two new alkaloid glycosides were obtained, and their structures were identified by mass spectrometry and NMR data as(S)-7-hydroxy-1-(p-hydroxybenzyl)-2,2-N,N-dimethyl-1,2,3,4-tetrahydroisoquinoline-6-O-β-D-glucopyranoside and(S)-N-methyltetrahydropalmatubine-9-O-β-D-glucopyranoside, respectively. This study is of great significance for enriching the information about the chemical composition and the in-depth development of C. chinensis. Meanwhile, it can provide a reference for rapid identification and isolation of alkaloid glycosides from other Chinese herbal medicines.
		                        		
		                        		
		                        		
		                        			Glycosides/chemistry*
		                        			;
		                        		
		                        			Medicine, Chinese Traditional
		                        			;
		                        		
		                        			Tandem Mass Spectrometry/methods*
		                        			;
		                        		
		                        			Coptis chinensis
		                        			;
		                        		
		                        			Drugs, Chinese Herbal/chemistry*
		                        			;
		                        		
		                        			Alkaloids/analysis*
		                        			;
		                        		
		                        			Plant Extracts/chemistry*
		                        			;
		                        		
		                        			Antineoplastic Agents
		                        			;
		                        		
		                        			Plants, Medicinal/chemistry*
		                        			;
		                        		
		                        			Water
		                        			;
		                        		
		                        			Chromatography, High Pressure Liquid/methods*
		                        			;
		                        		
		                        			Coptis/chemistry*
		                        			
		                        		
		                        	
7.The crucial toxic components of ambient fine particles promoted the maturation and differentiation of megakaryocytes.
Li Ting XU ; Ze ZHANG ; Hai Yi YU ; Xiao Ting JIN ; Yu Xin ZHENG
Chinese Journal of Preventive Medicine 2022;56(9):1314-1322
		                        		
		                        			
		                        			Objective: To reveal the crucial toxic components of ambient fine particles (PM2.5) that affect the maturation and differentiation of megakaryocytes. Methods: Human megakaryocytes were exposed to the organic fractions, metallic fractions and water-soluble fractions of PM2.5 at two exposure doses (i.e. actual air proportion concentration or the same concentration), respectively. The cell viability was performed to screen the non-cytotoxic levels of toxic components of PM2.5 using the CCK-8 assay. CellTiter-Blue assay, morphological observation, flow cytometry analysis and WGA staining assay were used to evaluate the cell morphological changes, occurrence of DNA ploidy, alteration in the expressions of biomarkers and platelet formation, which were key indicators of the maturation and differentiation of megakaryocytes. Results: Compared to the control group, both metallic and organic components of PM2.5 resulted in a lag in megakaryocytes with an increase in cell volume and the onset of DNA ploidy. Flow cytometry analysis showed that CD33 (the marker of myeloid-specific) decreased and CD41a (a megakaryocyte maturation-associated antigen) increased in metallic and organic components of PM2.5 treatment groups. Moreover, compared to the control group, budding protrusions increased in metallic and organic components of PM2.5 treatment groups. The water-soluble components had no effect on the maturation and differentiation of macrophages. Conclusion: Metallic and organic components of PM2.5 are the crucial toxic components that promote the maturation and differentiation of megakaryocytes.
		                        		
		                        		
		                        		
		                        			Biomarkers
		                        			;
		                        		
		                        			DNA/pharmacology*
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Megakaryocytes/chemistry*
		                        			;
		                        		
		                        			Particulate Matter/toxicity*
		                        			;
		                        		
		                        			Sincalide/pharmacology*
		                        			;
		                        		
		                        			Water/pharmacology*
		                        			
		                        		
		                        	
8.Quality control of substance benchmarks for Danggui Sini Decoction.
Chen-Na LU ; Xiao YE ; Xiao-Qian LIU ; Wei-Hong FENG ; Yao-Hua LIANG ; Chun LI ; Zhi-Min WANG
China Journal of Chinese Materia Medica 2022;47(15):4007-4014
		                        		
		                        			
		                        			A comprehensive quality control method was established to provide references for quality control and evaluation of substance benchmarks of Danggui Sini Decoction(DSD). The HPLC separation was performed on a Kromasil 100 C-8 column(4.6 mm×250 mm, 5 μm) with acetonitrile(A)-0.05% phosphoric acid in water(B) as mobile phase in a gradient elution mode at the flow rate of 1 mL·min~(-1). The column temperature was 25 ℃ and the detection wavelength was set at 275 nm. Under these conditions, the content of seven components, including paeoniflorin, liquiritin, cinnamic acid, cinnamaldehyde, ammonium glycyrrhetate, ligustilide, and asarinin was simultaneously determined. Under the same chromatographic conditions, the HPLC fingerprint method for analysis of 15 batches of DSD was established. The content determination of aristolochic acid I, using the same test solution as the content determination item, was performed on an ACQUITY UPLC BEH C_(18) column(2.1 mm×50 mm, 1.7 μm) with methanol(A)-water(including 0.1% formic acid and 5 mmol·L~(-1) ammonium formate)(B) as the mobile phase in a gradient elution mode at the flow rate of 0.4 mL·min~(-1) and the column temperature of 40 ℃ by LC-MS/MS. The MS conditions included electrospray ionization(ESI) as an ion source, positive ion ionization, selective reaction monitoring(SRM), the parent ion of 359.3, and the daughter ion of 297.8. The results of the methodological investigation all met the requirements of content determination/fingerprint determination. As a result, the content ranges of paeoniflorin, liquiritin, cinnamic acid, cinnamaldehyde, ammonium glycyrrhetate, ligustilide, and asarinin were 5.419 8-11.267 3, 1.023-3.669 8, 0.145 6-0.444 1, 0.099 1-0.321 9, 3.159 1-7.731 9, 0.146 4-0.471 7, and 0.237 3-0.401 0 mg·g~(-1), respectively. Twenty-two common peaks were selected and 10 of them were identified by the comparison with the reference substances. The fingerprint similarity of 15 batches of DSD was in the range of 0.91-0.996 and the content of aristolochic acid I in DSD was 300.03-638.13 ng·g~(-1). The method established in this study is reliable and easy to operate and has great practical value, which can be used for overall quality control of substance benchmarks for DSD.
		                        		
		                        		
		                        		
		                        			Ammonium Compounds
		                        			;
		                        		
		                        			Benchmarking
		                        			;
		                        		
		                        			Chromatography, High Pressure Liquid/methods*
		                        			;
		                        		
		                        			Chromatography, Liquid
		                        			;
		                        		
		                        			Drugs, Chinese Herbal/chemistry*
		                        			;
		                        		
		                        			Quality Control
		                        			;
		                        		
		                        			Tandem Mass Spectrometry/methods*
		                        			;
		                        		
		                        			Water
		                        			
		                        		
		                        	
9.Pharmacodynamic material basis and anti-inflammatory mechanism of Chrysanthemum morifolium cv. Fubaiju based on UPLC-Q-TOF-MS/MS combined with network pharmacology.
Shi-Qin WANG ; Dan LIU ; Xiao-Chuan YE ; Qian-Qian ZHU ; Dan-Dan ZHANG ; Bo WANG ; Jing NIE
China Journal of Chinese Materia Medica 2022;47(15):4190-4201
		                        		
		                        			
		                        			The chemical components in rats after oral administration of the water extract of Chrysanthemum morifolium cv. Fubaiju(CMF) were analyzed by UPLC-Q-TOF-MS/MS technique. Forty-four compounds were identified from the water extract of CMF and 11 components were identified from the rat serum. A total of 264 potential anti-inflammatory targets were identified by network pharmacology based on serum components. The "component-target" network and protein-protein interaction(PPI) network were constructed, and GO function enrichment and KEGG pathway enrichment analyses were performed. The molecular docking was carried out to validate the results of network pharmacology. The results showed that CMF might act on AKT1, TNF, TP53, IL6, INS, and other core targets through apigenin, luteolin, acacetin, diosmetin, 3,4-O-dicaffeoylquinic acid, and other active components, and exert anti-inflammatory effects by regulating PI3 K-AKT signaling pathway, FoxO signaling pathway, cAMP signaling pathway, Ras signaling pathway, and other pathways. The pharmacodynamic materials basis of CMF was identified by UPLC-Q-TOF-MS/MS technology, and the core anti-inflammatory targets and the underlying mechanism of action were analyzed by network pharmacology and molecular docking, which provided a reference for comprehensively clarifying the pharmacodynamic materials basis and quality control of CMF.
		                        		
		                        		
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Anti-Inflammatory Agents/pharmacology*
		                        			;
		                        		
		                        			Chrysanthemum
		                        			;
		                        		
		                        			Drugs, Chinese Herbal/chemistry*
		                        			;
		                        		
		                        			Molecular Docking Simulation
		                        			;
		                        		
		                        			Network Pharmacology
		                        			;
		                        		
		                        			Rats
		                        			;
		                        		
		                        			Tandem Mass Spectrometry/methods*
		                        			;
		                        		
		                        			Water
		                        			
		                        		
		                        	
10.Study on drug properties of Arisaematis Rhizoma and Arisaema Cum Bile based on substance and energy metabolism in normal and cold/heat syndrome model rats.
Fa-Zhi SU ; Chen-Xi BAI ; Wen-Sen ZHANG ; Jing YANG ; Jian-Ping HU ; Yan-Ping SUN ; Bing-You YANG ; Hai-Xue KUANG ; Qiu-Hong WANG
China Journal of Chinese Materia Medica 2022;47(17):4682-4690
		                        		
		                        			
		                        			This paper clarified the scientific connotation of the changes in cold and heat properties of Arisaematis Rhizoma and Arisaema Cum Bile through investigating the changes of substance and energy metabolism after drug intervention in the rats with normal and cold/heat syndrome, so as to improve the method of evaluating the drug properties of Chinese medicine. After one week of adaptive feeding, healthy male SD rats were randomly divided into three parts: normal rats, heat syndrome rat models, and cold syndrome rat models. Through ice water bath and oral euthyrox(120 μg·kg~(-1)), the models of cold syndrome and heat syndrome were induced, respectively. The models were made at 9:00 am. and administrated by gavage at 3:00 pm. every day. All administration groups were administrated with Arisaematis Rhizoma and Arisaema Cum Bile decoction, respectively, and the blank group was given the same dose of normal saline. After continuous administration for 15 d, the rats were anesthetized by chloral hydrate, blood was taken from abdominal aorta, and the hearts and livers were removed and stored at-80 ℃. The changes in the body weight and anal temperature of rats during administration were detected, and the liver coefficient of rats was detected after removing the liver. Enzyme-linked immunosorbent assay(ELISA) was adopted to detect the expression level of the indexes related to substance and energy metabolism in liver and heart of rat, and Western blot was used to detect the expression of key proteins in AMPK/mTOR signaling pathway for further verification. The results showed that Arisaematis Rhizoma enhanced the expression level of enzymes related to substance and energy metabolism in the normal and cold and heat syndrome rat models, and increased anal temperature, which exhibited warm(hot) drug property. Arisaema Cum Bile inhibited the level of substance and energy metabolism in rats, and reduced anal temperature, which showed cold(cool) drug property. Chinese Pharmacopoeia has recorded "Arisaematis Rhizoma has warm property and Arisaema Cum Bile has cool property", which is consistent with the phenomenon in this study. Therefore, it is feasible to evaluate the drug properties of Chinese medicine based on the substance and energy metabolism of normal and cold/heat syndrome model rats, which completes the method of evaluating drug properties of Chinese medicine.
		                        		
		                        		
		                        		
		                        			AMP-Activated Protein Kinases
		                        			;
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Arisaema/chemistry*
		                        			;
		                        		
		                        			Bile
		                        			;
		                        		
		                        			Chloral Hydrate
		                        			;
		                        		
		                        			Cold-Shock Response/drug effects*
		                        			;
		                        		
		                        			Drugs, Chinese Herbal/therapeutic use*
		                        			;
		                        		
		                        			Energy Metabolism
		                        			;
		                        		
		                        			Heat Stroke/therapy*
		                        			;
		                        		
		                        			Hot Temperature
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Rats
		                        			;
		                        		
		                        			Rats, Sprague-Dawley
		                        			;
		                        		
		                        			Saline Solution
		                        			;
		                        		
		                        			Syndrome
		                        			;
		                        		
		                        			TOR Serine-Threonine Kinases
		                        			;
		                        		
		                        			Thyroxine
		                        			;
		                        		
		                        			Water
		                        			
		                        		
		                        	
            
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