Toxic effects of Tripterygium glycoside tablets on the reproductive system of male rats by metabolomics, cytotoxicity, and molecular docking.

BACKGROUND: Tripterygium glycoside tablets (TGT) is the most common preparation from Tripterygium wilfordii Hook F, which is widely used in clinical for treating rheumatoid arthritis (RA) and other autoimmune diseases. However, its serious reproductive toxicity limits its application. PURPOSE: This study aimed to elucidate the toxic effects of TGT on the reproductive system of male RA rats and its potential toxic components and mechanism. METHODS: Collagen-induced arthritis (CIA) rat model was established, and TGT suspension was given at low, medium, and high doses. Gonadal index, pathological changes, and the number of spermatogenic cells were used to evaluate the toxic effects of TGT on the reproductive system. Non-targeted metabolomics of testicular tissue was conducted by UHPLC-QTOF/MS. Combined with network toxicology, the key targets of TGT-induced reproductive toxicity were screened and RT-qPCR was used to validation. In vitro toxicity of 19 components of TGT was evaluated using TM3 and TM4 cell lines. Molecular docking was used to predict the interaction between toxic components and key targets. RESULTS: TGT reduced testicular and epididymis weight. Pathology analysis showed a lot of deformed and atrophic spermatogenic tubules. The number of spermatogenic cells decreased significantly (P<0.0001). A total of 58 different metabolites including platelet-activating factor (PAF), lysophosphatidylcholine (Lyso PC), phosphatidylinositol (PI), glutathione (GSH), and adenosine monophosphate (AMP) were identified by testicular metabolomics. Glycerophospholipid metabolism, ether lipid metabolism, and glutathione metabolism were key pathways responsible for the reproductive toxicity of TGT. Ten key reproductive toxicity targets were screened by network toxicology. The cytotoxicity test showed that triptolide, triptonide, celastrol, and demethylzeylasteral could significantly reduce the viability of TM3 and TM4 cells. Alkaloids had no apparent toxic effects. Molecular docking showed that the four toxic components had a good affinity with 10 key targets. All binding energies were less than -7 kcal/mol. The RT-qPCR results showed the Cyp19a1 level was significantly up-regulated. Pik3ca and Pik3cg levels were significantly down-regulated. CONCLUSION: Through testicular metabolomics, we found that TGT may cause reproductive toxicity through CYP19A1, PIK3CA, and PIK3CG three target, which was preliminarily revealed. This study laid the foundation for elucidating the toxicity mechanism of TGT and evaluating its safety and quality.

[Soil physical and chemical properties and vegetation characteristics of different types of grassland in Qilian Mountains, China]. / 祁连山不同类型草地的土壤理化性质与植被特征.

Grasslands in Qilian Mountains plays an important role in maintaining the ecological security of western China. To understand soil physical and chemical properties and the distribution characteristics of vegetation, as well as their correlation in different types of grasslands in Qilian Mountains, we measured soil moisture, nutrient content, bulk density, particle composition, and vegetation characteristics in seven types of grassland in Qilian Mountains. The fractal dimension of soil particles, soil organic carbon, total nitrogen and total phosphorus storages in 0-40 cm soil layer, and plant diversity index were calculated. The results showed that there were significant differences in soil physical and chemical properties and vegetation characteristics among different grassland types. Compared with other types of grassland, alpine meadow had higher soil water, nutrient and clay content, but lower bulk density and sand content. Soil organic carbon, total nitrogen and total phosphorus storages in 0-40 cm layer ranged from 3084 to 45247, 164 to 2358 and 100 to 319 g·m-2, respectively, with high contents of organic carbon and total nitrogen and low content of total phosphorus. There was a significant positive correlation between soil total phosphorus storage and plant diversity index, indicating that soil total phosphorus content was the key factor affec-ting grassland plant diversity in Qilian Mountains. Compared with other grassland types, alpine meadow in Qilian Mountains had better vegetation status, soil moisture, and nutrient conditions.

Potential hepatoxicity risk of the shell of Herpetospermum caudigerum Wall in rats based on 1H-NMR metabonomics.

The Herpetospermum caudigerum Wall (HCW) is a traditional Tibetan medicine and is widely used in clinical practice. However, the shell of the HCW (SHCW) has rarely been studied, and some researchers have suggested that the SHCW may be toxic. Therefore, in this study, SHCW was administered to rats at two doses (0.1 and 0.33 g/kg) once a day for 21 days. The hepatic stimuli induced by SHCW in rats were investigated for the first time by 1H-NMR-based metabolomics combined with histopathological observation and biochemical detection. Histopathological sections showed a certain degree of hepatocyte edema and hepatic sinus congestion in the liver tissue of the rats in the drug-administered group. Serum biochemical indicators revealed a significant increase in ALT, AST, and MDA, and a significant decrease in SOD. Metabolomic results showed that the metabolites in rats were changed after gavage administration of extracts from SHCW. By multivariate statistical analysis and univariate analysis, it was found that SHCW could cause the disorder of energy metabolism, oxidative stress and amino acid metabolism in rats, leading to liver damage. This comprehensive metabolomics approach demonstrates its ability to describe the global metabolic state of an organism and provides a powerful and viable tool for exploring drug-induced toxicity or side effects.

Hepatoprotection of Herpetospermum caudigerum Wall. against CCl4-induced liver fibrosis on rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Herpetospermum caudigerum Wall. (HCW) is a traditional Tibetan medicine, which has been used to ameliorate liver injuries in the folk. AIM OF THE STUDY: Liver fibrosis has been recognized as a major lesion of the liver that leads to liver cirrhosis/hepatocarcinoma and even to death in the end. This study aims to demonstrate the protective effect of HCW against CCl4-induced liver injury in rats and to explore the underlying mechanisms. MATERIALS AND METHODS: Hepatic fibrosis was induced by intraperitoneal injection of CCl4. Liver function markers, fibrosis markers, serum anti-oxidation enzymes as well as elements levels were determined. Serum and liver tissues were subjected to NMR-based metabolomics and multivariate statistical analysis. RESULTS: HCW could significantly reduce the elevated levels of fibrosis markers such as hyaluronidase, laminin, Type III procollagen and Type IV collagen in the serum, improve the activities of the antioxidant enzymes, and effectively reverse the abnormal levels of elements in liver fibrosis rats. Correlation network analysis revealed that HCW could treat liver fibrosis by ameliorating oxidative stress, repairing the impaired energy metabolisms and reversing the disturbed amino acids and nucleic acids metabolisms. CONCLUSION: This integrated metabolomics approach confirmed the validity of the traditional use of HCW in the treatment of liber fibrosis, providing new insights into the underlying mechanisms.

Hepatoprotective effect of Herpetospermum caudigerum Wall. on carbon tetrachloride-induced hepatic fibrosis in rats.

Tibetan medicine Herpetospermum caudigerum Wall. (HCW) has long been employed to treat hepatitis, inflammatory diseases and jaundice according to the records of "The Four Medical Tantras" in China. This study was investigated to explore the protective effects of HCW on hepatic fibrosis and the possible mechanism in a rat model. Hepatic fibrosis was established by intragastric administration of 3 ml/kg carbon tetrachloride (CCl4 ) twice a week for 6 weeks. CCl4 -treated rats were received HCW (1 and 3 g/kg/d) and silymarin (0.1 g/kg/d) from 3 to 6 weeks. The results showed that HCW could significantly decrease the levels of AST, ALT, HA, LN, PCIII, Col IV, TNF-α, IL-1ß and IL-6. Moreover, HCW could effectively inhibit collagen deposition and reduce the pathological damage. Analysis experiments finally exhibited that HCW was able to markedly inhibit hepatic fibrosis by modulating the expressions of NF-κB p65, IκBα, Samd3 and TGF-ß1 proteins. Therefore, our results suggest that HCW has hepatoprotective activity against CCl4 -induced hepatic fibrosis in rats by regulating the inflammatory responses.

A label-free assay for T4 polynucleotide kinase/phosphatase activity and its inhibitors based on poly(thymine)-templated copper nanoparticles.

DNA 3'-phosphatase takes an important role in DNA damage repair, replication and recombination. Here, we present a novel label-free fluorescent assay for T4 polynucleotide kinase/phosphatase (T4 PNKP) activity and its inhibitor screening by using poly(thymine)-templated fluorescent copper nanoparticles (CuNPs) as a fluorescent indicator. In this assay, we designed a simple T-rich hairpin primer with a 3'-phosphoryl end, which can serve as both the substrate for T4 PNKP and DNA template for the formation of fluorescent CuNPs. Once the phosphorylated hairpin primer was hydrolyzed by T4 PNKP, the resulting hairpin primer with a 3'-hydroxyl end was immediately elongated to form a long double-strand product by DNA polymerase, which prohibited the formation of fluorescent CuNPs due to the lack of poly T single-stranded DNA template. This new strategy provides a sensitive, selective, and cost-effective manner for T4 PNKP analysis, which holds a great potential in the study of DNA damage repair mechanisms.

[Study on methodology for evaluating clinical efficacy of traditional Tibetan medicine].

The evaluation on clinical efficacy of traditional Tibetan medicine (TTM) is an important scientific subject during the development of TTM. Firstly, the authors introduced the current situations and problems in evaluation on clinical efficacy of traditional Tibetan medicine both at home and abroad in this study. Secondly, they compared the similarities and differences between TTM and traditional Chinese medicine (TCM) in evaluation on clinical efficacy, define their differences in details but not in nature, and proposed that TTM could selectively learn TCM's experiences in clinical research and build a specific methodology system for evaluation on clinical efficacy according to its own characteristics. Thirdly, they discussed the methodological challenges in evaluation on clinical efficacy of TTM, including the pending clinical research guidelines and disease diagnosis standards according to its own characteristics. Finally, they propound some suggestions for promoting the evaluation on clinical efficacy of TTM, including the comprehensive application of multiple research methods, overall research-based evaluation on efficacy of TTM complex intervention and selection of accepted and objective outcome indexes for efficacy evaluation.

[Determination of eight heavy metals and two main ingredients of safflower planted in linzhi of Tibetan].

The eight heavy metals and two essential constitutes of safflowers planted in linzhi which lies in Southern Tibet were analyzed by ICP-MS and by HPLC respectively. Heavy metals of safflower in the region were at the lower level and the essential constitutes were at the higher level. The better quality of safflower here was assisted by the excellent climate in tibet.

Study on methodology for evaluating clinical efficacy of traditional Tibetan medicine / 中国中药杂志

The evaluation on clinical efficacy of traditional Tibetan medicine (TTM) is an important scientific subject during the development of TTM. Firstly, the authors introduced the current situations and problems in evaluation on clinical efficacy of traditional Tibetan medicine both at home and abroad in this study. Secondly, they compared the similarities and differences between TTM and traditional Chinese medicine (TCM) in evaluation on clinical efficacy, define their differences in details but not in nature, and proposed that TTM could selectively learn TCM's experiences in clinical research and build a specific methodology system for evaluation on clinical efficacy according to its own characteristics. Thirdly, they discussed the methodological challenges in evaluation on clinical efficacy of TTM, including the pending clinical research guidelines and disease diagnosis standards according to its own characteristics. Finally, they propound some suggestions for promoting the evaluation on clinical efficacy of TTM, including the comprehensive application of multiple research methods, overall research-based evaluation on efficacy of TTM complex intervention and selection of accepted and objective outcome indexes for efficacy evaluation.

Determination of eight heavy metals and two main ingredients of safflower planted in linzhi of Tibetan / 中国中药杂志

The eight heavy metals and two essential constitutes of safflowers planted in linzhi which lies in Southern Tibet were analyzed by ICP-MS and by HPLC respectively. Heavy metals of safflower in the region were at the lower level and the essential constitutes were at the higher level. The better quality of safflower here was assisted by the excellent climate in tibet.

Vitamin D intake and type 2 diabetes risk: a meta-analysis of prospective cohort studies.

BACKGROUND: The findings form studies on the relationship between vitamin D and type 2 diabetes were inconsistent. OBJECTIVES: To elucidate the association between vitamin D consumption and type 2 diabetes risk by conducting a meta-analysis. METHODS: We conducted a systematic literature search to identify prospective cohort studies of vitamin D intake and type 2 diabetes risk prior to November 2012. Eligible studies were retrieved via both computer searches and manual review of references. The summary risk estimates were calculated based on the highest versus the lowest categories. RESULTS: Meta-analysis of 4 prospective cohort studies involving 187, 592 participants and 9, 456 incident cases showed an absence of significant association between total vitamin D intake and type 2 diabetes risk. The combined RR was 0.93 (95% CI: 0.85-1.01). The associations were similar for subgroup analyses, a combined RR respectively was 0.94 (95% CI: 0.77-1.08), 0.91 (95% CI: 0.77-1.08), 0.93 (95% CI: 0.84-1.02), and 0.92 (95% CI: 0.84-1.01) for the intake of dietary vitamin D, supplemental vitamin D, total vitamin D in USA and total vitamin D for women only. CONCLUSIONS: Our results support that there was no association between vitamin D intake and type 2 diabetes.

[Improvement in blood glucose fluctuations of insulin-treated type 1 diabetic patients with additional use of acarbose and assessed by continuous blood glucose monitoring program].

OBJECTIVE: To investigate the effect of combined use of insulin and acarbose on glucose excursion in type 1 diabetic patients. METHODS: 120 cases were randomly divided into control group and observation group. The control group received preprandial ultra-short effect insulin and long-acting insulin before bedtime while the observation group received acarbose 50 mg added to the medicine taken by the control group. Continuous Glucose Monitoring System (CGMS) was used to watch the blood glucose fluctuations. Data related to blood glucose level, glucose excursions after meals and hypoglycemia at night were compared between patients in the two groups. RESULTS: The average blood glucose (9.37 ± 1.70) mmol/L, the largest amplitude of glycemic excursions (LAGE) (11.42 ± 2.73) mmol/L, hyperglycemia-area under curve 0.89 ± 0.54, mean amplitude of glycemic excursions (MAGE) (5.13 ± 2.23) mmol/L, M-value (18.93 ± 11.43) mmol/L and insulin dosage (42.11 ± 14.42) U/day of observation group were significantly lower than in the control group (P < 0.05). Glucose excursions after meals and the times (0.33 ± 0.50)/day, the maintenance time (43.75 ± 43.50)/min and low glycemic index (LBGI) (0.005 ± 0.002) mmol/L of hypoglycemia at night were also significantly lower than in the control group, with statistically significant (P < 0.05) differences. CONCLUSION: The blood glucose fluctuation was significantly improved, with the decrease of insulin dosage while both glucose excursions and hypoglycemia at night reduced in patients with type1 diabetes mellitus after the acarbose treatment.We suggested that this program deserve further observation.

Xylocoside G reduces amyloid-ß induced neurotoxicity by inhibiting NF-κB signaling pathway in neuronal cells.

Amyloid-ß (Aß) peptide, which can invoke a cascade of inflammatory responses, is considered to play a causal role in the development and progress of Alzheimer's disease (AD). Xylocoside G (XG) is an active compound isolated from a traditional Chinese medicinal plant, Itoa orientalis. We have previously reported that XG has neuroprotective effects, of which the mechanism is yet unknown. In this study, we investigated the possible mechanisms underlying neuroprotection of XG against Aß-induced toxicity in SH-SY5Y cells and primary neurons. Pretreatment with XG significantly attenuated the cell viability reduction induced by Aß exposure in a dose dependent manner which was testified by 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase release assay. In addition, pretreatment with XG countered the effect of Aß on Bax and Bcl-2 expression and repressed Aß-induced caspase-3 activation, suggesting that the neuroprotective effect of XG is associated with apoptosis regulation. Neuroinflammation has been implicated in Aß-induced neuronal death. XG significantly attenuated Aß-stimulated release of inflammatory factors such as tumor necrosis factor-α, interleukin-1ß, and prostaglandin E2. It also downregulated the expression of cyclooxygenase-2 in SH-SY5Y cells. Further molecular mechanism studies demonstrated that XG inhibited Aß-induced NF-κB p65 translocation, which was probably the result of inhibition of JNK phosphorylation but not ERK or p38 MAPK pathway by XG. This is the first study to demonstrate that XG protects SH-SY5Y cells against Aß-induced inflammation and apoptosis by down-regulating NF-κB signaling pathways.