Quality evaluation of compounds in leaves of six Taxus species based on UPLC-MS/MS and chemometrics.

Introduction: Taxus species are used as medicinal plants all over the world. The leaves of Taxus species are sustainable medicinal resources that are rich in taxoids and flavonoids. However, traditional identification methods cannot effectively identify Taxus species on the basis of leaces used as raw medicinal materials, because their appearance and morphological characteristics are almost the same, and the probability of error identification increases in accordance with the subjective consciousness of the experimenter. Moreover, although the leaves of different Taxus species have been widely used, their chemical components are similar and lack systematic comparative research. Such a situation is challenging for quality assessment. Materials and methods: In this study, ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry combined with chemometrics was applied for the simultaneous determination of eight taxoids, four flavanols, five flavonols, two dihydroflavones, and five biflavones in the leaves of six Taxus species, namely, T. mairei, T. chinensis, T. yunnanensis, T. wallichiana, T. cuspidata, and T. media. Chemometric methods, including hierarchical cluster analysis, principal component analysis, orthogonal partial least squares-discriminate analysis, random forest iterative modeling, and fisher linear discriminant analysis, were utilized to differentiate and evaluate the six Taxus species. Results: This proposed method exhibited good linearity (R 2 = 0.9999-0.9972) with a lower quantification limits of 0.94-3.05 ng/mL for all analytes. The intra- and inter-day precisions were within 6.83%. Six compounds, namely, 7-xylosyl-10-deacetyltaxol, ginkgetin, rutin, aromadendrin, 10-deacetyl baccatin III, and epigallocatechin, were identified through chemometrics for the first time. These compounds can be used as important chemical markers to distinguish the above six Taxus species rapidly. Conclusion: This study established a method for determination of the leaves of six Taxus species, and revealing the differences in the chemical components of these six Taxus species.

Aucubin enhances the antitumor activity of cisplatin through the inhibition of PD-L1 expression in hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is a leading cause of cancer-associated mortality in the world. However, the anticancer effects of aucubin against HCC have yet to be reported. Cisplatin often decreased CD8+ tumor-infiltrating lymphocytes in the tumor microenvironment through increasing programmed death-ligand 1 (PD-L1) expression, which seriously affected the prognostic effect of cisplatin in the treatment of patients with HCC. Therefore, it is necessary to identify a novel therapeutic avenue to increase the sensitivity of cisplatin against HCC. PURPOSE: This study aims to evaluate the anti-tumor effect of aucubin on HCC, and also to reveal the synergistic effects and mechanism of aucubin and cisplatin against HCC. STUDY DESIGN AND METHODS: An H22 xenograft mouse model was established for the in vivo experiments. Cancer cell proliferation was detected by MTT assay. RT-qPCR was performed to analyze CD274 mRNA expression in vitro. Western blotting was employed to determine the expression levels of the PD-L1, p-Akt, Akt, p-ß-catenin, and ß-catenin in vitro. Immunofluorescence was carried out to examine ß-catenin nuclear accumulation in HCC cells. Immunohistochemistry was used to detect tumoral PD-L1 and CD8α expression in xenograft mouse model. RESULTS: Aucubin inhibits tumor growth in a xenograft HCC mouse model, but did not affect HCC cell viability in vitro. Aucubin treatment significantly inhibited PD-L1 expression through inactivating Akt/ß-catenin signaling pathway in HCC cells. Overexpression of PD-L1 dramatically reversed aucubin-mediated tumoral CD8+ T cell infiltration and alleviated the antitumor activity of aucubin in xenograft mouse model. Moreover, Cisplatin could induce the expression of PD-L1 through the activation of the Akt/ß-catenin signaling pathway in HCC cells, which can be blocked by aucubin in vitro. In xenograft mouse model, cisplatin treatment induced PD-L1 expression and alleviated the infiltration of CD8+ T lymphocytes in the tumor microenvironment. Aucubin not only abrogated cisplatin-induced PD-L1 expression but also enhanced the antitumor efficacy of cisplatin in a mouse xenograft model of HCC. CONCLUSION: Aucubin exerts antitumor activity against HCC and also enhances the antitumor activity of cisplatin by suppressing the Akt/ß-catenin/PD-L1 axis.

Lycium barbarum polysaccharide protects against osteonecrosis of femoral head via regulating Runx2 expression.

BACKGROUND: Osteonecrosis of femoral head (ONFH) is a pathological state caused by lack of blood supply in femoral head. This study aimed to explore the function of Lycium barbarum polysaccharide (LBP), an antioxidant agent extracted from L. barbarum, on ONFH. METHODS: Osteonecrosis rat model was generated using lipopolysaccharide (LPS) and methylprednisolone followed by examination of body weight, blood glucose, morphology, and BMSC osteoblast differentiation. The effect and underlying mechanism of LBP on the proliferation, apoptosis, and osteoblast differentiation of BMSC were determined with or without LPS or hypoxia treatment using CCK-8. Alizarin Red S staining, flow cytometry, and western blot, respectively. RESULT: LBP could protect against glucocorticoid-induced ONFH in rats, resulting in improved sparse trabecular bone, empty lacunae and bone cell coagulation. Moreover, LBP promoted the proliferation and osteoblast differentiation of bone mesenchymal-derived stem cells (BMSCs) in a dose-dependent manner. Furthermore, LBP enhanced osteoblast differentiation of BMSCs under hypoxia condition. Mechanistically, we found that LBP treatment enhanced Runx2 and ALP expression in BMSCs. LBP restored the expression of Runx2 and ALP under hypoxia, suggesting that LBP might be involved in regulating Runx2/ALP expression and contributed to osteoblast differentiation. Knockdown of Runx2 significantly inhibited BMSCs proliferation, while LBP treatment did not rescue the osteoblast differentiation ability of BMSCs with Runx2 knockdown. CONCLUSION: Our findings suggested that LBP protects against ONFH via regulating Runx2 expression, which could be utilized to treat patients suffering ONFH.

Antitumor and anti-inflammatory effects of oligosaccharides from Cistanche deserticola extract on spinal cord injury.

In the present study, the pharmacological effects of oligosaccharides from Cistanche deserticola extract on inflammation, oxidative stress, and apoptosis in male albino rats with spinal cord injury were investigated. Lipid peroxidation, GSH, catalase, superoxide dismutase, acetylcholine esterase, GPx, ROS, and nitric acid were significantly altered in the rats with spinal cord injury. The mRNA expression levels of IL-6, TNF-α, cyclooxygenase-2, iNOS, p53, caspase-3, bax, and pro-NGF were reduced by >20% following extract supplementation. Protein expression levels of caspase-3 and pro-NGF were also reduced by >20%. The number of p53 positive cells was 1, 79, 54, 33, and 19 in groups GI-GV, respectively, and the corresponding numbers of caspase-3 positive cells were 2, 87, 51, 23, and 14. Based on the present results, the use of oligosaccharides from Cistanche deserticola extract was effective against inflammation, oxidative stress, and apoptosis in spinal cord injury male albino rats.

Comparing the antidiabetic effects and chemical profiles of raw and fermented Chinese Ge-Gen-Qin-Lian decoction by integrating untargeted metabolomics and targeted analysis.

BACKGROUND: Microbial fermentation has been widely applied in traditional Chinese medicine (TCM) for thousands of years in China. Various beneficial effects of fermentation for applications in TCM or herbals have been reported, such as enhanced anti-ovarian cancer, antioxidative activity, and neuroprotective effects. Ge-Gen-Qin-Lian decoction (GQD), a classic TCM formula, has been used to treat type 2 diabetes mellitus in China. In this study, GQD was fermented with Saccharomyces cerevisiae, and the antidiabetic activities and overall chemical profiles of raw and fermented GQD (FGQD) were systematically compared. METHODS: First, the antidiabetic effects of GQD and FGQD on high-fat diet and streptozotocin (STZ)-induced diabetic rats were compared. Then, high-performance liquid chromatography Q Exactive MS was applied for rapid characterization of the chemical components of GQD. Additionally, we proposed an integrated chromatographic technique based untargeted metabolomics identifying differential chemical markers between GQD and FGQD and targeted analysis determining the fermenting-induced quantitative variation tendencies of chemical marker strategy for overall chemical profiling of raw and fermented GQD. RESULTS: Both GQD and FGQD displayed effects against HFD and STZ-induced diabetes, and FGQD showed a better recovery trend associated with profound changes in the serum lipoprotein profile and body weight gain. In addition, 133 compounds were characterized from GQD. It was demonstrated that the integrated strategy holistically illuminated 30 chemical markers contributed to the separation of GQD and FGQD, and further elucidated the fermenting-induced chemical transformation mechanisms and inherent chemical connections of secondary metabolites. Although there were no new secondary metabolites in FGQD compared with GQD, the amounts of secondary metabolites, which were mostly deglycosylated, were redistributed in FGQD. CONCLUSION: The anti-diabetic activities of GQD could be improved by applying fermentation technology. Moreover, the proposed strategy could serve as a powerful tool for systematically exploring the chemical profiles of raw and fermented formulas.

Zuo Gui Wan Alters Expression of Energy Metabolism Genes and Prevents Cell Death in High-Glucose Loaded Mouse Embryos.

BACKGROUND: Zuo Gui Wan (ZGW) is a classic formula in traditional chinese medicine (TCM). Previous studies have shown that it is beneficial for impaired glucose tolerance (IGT) of adults and the offspring as well. This study aimed to understand the molecular mechanisms of the efficacy of ZGW on IGT. METHODS: We used high-glucose loaded 2-cell stage mouse embryos as a model and took advantage of single-cell RNA sequencing technology to analyze the transcriptome of the model with or without ZGW. Differential gene expression analysis was performed with DESeq2. RESULTS: High glucose can downregulate genes in the ribosome pathway, while ZGW can reverse this inhibition and as a result prevent embryo cell death caused by high glucose. Furthermore, high glucose can affect sugar metabolism and influence mitochondrial function, but ZGW can promote sugar metabolism via the tricarboxylic acid cycle mainly through upregulating the genes in the respiratory chain and oxidative phosphorylation. CONCLUSIONS: ZGW had a protective effect on embryonic cell death caused by glucose loading. The reversion of inhibition of ribosome pathway and regulation of mitochondrial energy metabolism are main effects of ZGW on high-glucose loaded embryos. This research not only revealed the global gene regulation changes of high glucose affecting 2-cell stage embryos but also provided insight into the potential molecular mechanisms of ZGW on the IGT model.

[Treatment of Knee Osteoarthritis with Acupuncture plus Intra-articular Injection of Sodium Hyaluronate].

OBJECTIVE: To examine the clinical benefits of acupuncture combined with intra-articular injection of sodium hyaluronate for knee osteoarthritis (KOA) patients. METHODS: A total of 150 KOA patients were randomized into simple medication and acupuncture plus medication (combined treatment) groups (n＝75 in each). For all the patients, intra-articular injection of sodium hyaluronate (2 mL) was performed once a week for 5 weeks, and for patients of the combined treatment group, filiform needles were separately inserted into unilateral or bilateral Zusanli (ST 36), Liangqiu (ST 34), Yanglingquan (GB 34), Yinlingquan (SP 9), Xiyangguan (GB 33), Dubi (ST 35), Neixiyan (EX-LE 4) and Xuehai (SP 10) according to the focus, and manipulated with the uniform reinforcing and reducing technique, followed by retaining the needles for 30 min. The acupuncture treatment was given once every day or every other day, 5 weeks in total. The visual analog scale (VAS) was used to assess the pain severity of knee-joint, and the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) employed to assess the 1) pain severity during various positions or movements (20 points), 2) severity of joint stiffness (8 points), and 3) difficulty in performing daily functional activities (68 points). The therapeutic effect was evaluated according to the decreased level of WOMAC subscale scores and improvement of daily activities. RESULTS: After the treatment, the VAS scores of both medication and combined treatment groups were decreased significantly in comparison with their own pre-treatment in each group (P<0.05), and that of the combined treatment group was significantly lower than that of the medication group (P<0.05). WOMAC and daily activity fin-dings showed that of the two 75 KOA patients in the medication and combined treatment groups, 16 (21.33%) and 32 (42.67%) experienced marked improvement, 46 (61.33%) and 38 (50.67%) were improved, and 13 (17.33%) and 5 (6.67%) ineffective, with the total effective rates being 82.67% and 93.33%, respectively. The therapeutic effect of the combined treatment group was apparently superior to that of the simple medication group (P<0.05). CONCLUSION: Acupuncture combined with intra-articular injection of sodium hyaluronate is effective in improving KOA patients' pain severity and other symptoms as well as functional activities.

Treatment of Knee Osteoarthritis with Acupuncture plus Intra-articular Injection of Sodium Hyaluronate / 针刺研究

OBJECTIVE: To examine the clinical benefits of acupuncture combined with intra-articular injection of sodium hyaluronate for knee osteoarthritis (KOA) patients. METHODS: A total of 150 KOA patients were randomized into simple medication and acupuncture plus medication (combined treatment) groups (n＝75 in each). For all the patients, intra-articular injection of sodium hyaluronate (2 mL) was performed once a week for 5 weeks, and for patients of the combined treatment group, filiform needles were separately inserted into unilateral or bilateral Zusanli (ST 36), Liangqiu (ST 34), Yanglingquan (GB 34), Yinlingquan (SP 9), Xiyangguan (GB 33), Dubi (ST 35), Neixiyan (EX-LE 4) and Xuehai (SP 10) according to the focus, and manipulated with the uniform reinforcing and reducing technique, followed by retaining the needles for 30 min. The acupuncture treatment was given once every day or every other day, 5 weeks in total. The visual analog scale (VAS) was used to assess the pain severity of knee-joint, and the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) employed to assess the 1) pain severity during various positions or movements (20 points), 2) severity of joint stiffness (8 points), and 3) difficulty in performing daily functional activities (68 points). The therapeutic effect was evaluated according to the decreased level of WOMAC subscale scores and improvement of daily activities. RESULTS: After the treatment, the VAS scores of both medication and combined treatment groups were decreased significantly in comparison with their own pre-treatment in each group (P<0.05), and that of the combined treatment group was significantly lower than that of the medication group (P<0.05). WOMAC and daily activity fin-dings showed that of the two 75 KOA patients in the medication and combined treatment groups, 16 (21.33%) and 32 (42.67%) experienced marked improvement, 46 (61.33%) and 38 (50.67%) were improved, and 13 (17.33%) and 5 (6.67%) ineffective, with the total effective rates being 82.67% and 93.33%, respectively. The therapeutic effect of the combined treatment group was apparently superior to that of the simple medication group (P<0.05). CONCLUSION: Acupuncture combined with intra-articular injection of sodium hyaluronate is effective in improving KOA patients' pain severity and other symptoms as well as functional activities.

Rivaroxaban in the Treatment of PICC-associated Upper Extremity Venous Thrombosis.

PURPOSE: Peripherally inserted central catheters (PICCs) are frequently used for prolonged drug administration, but their use is commonly complicated by the development of upper extremity deep venous thrombosis (UEDVT) requiring anticoagulation. Here, we compared the efficacy and safety profile of rivaroxaban (20 mg/d) with low molecular weight (LMW) heparin and vitamin K antagonists in the treatment of PICC-associated UEDVT. METHODS: Patients (N = 84) with PICC-associated UEDVT were studied. All had UEDVT identified by ultrasound scanning. Further ultrasound images were obtained at 1, 2, and 3 months after the start of treatment. Forty-four patients were treated with rivaroxaban and 40 with initial LMW heparin and vitamin K antagonist with continuation of vitamin K antagonists alone once international normalized ratio was therapeutic FINDINGS: In the rivaroxaban group mean (SD) age was 51 (16) years and 57% were men, whereas in the other group respective values were 50 (16) years and 56%. All patients were receiving treatment for cancer. Resolution of thrombus had occurred in 53.5% at 1 month, 76.1% at 2 months, and 92.6% at 3 months in the rivaroxaban-treated patients. Corresponding values in the LMW heparin/vitamin antagonist-treated patients were 34.2%, 55.5%, and 88.5%, respectively. Differences between groups were significant at 1 month (P < 0.01) and 2 months (P < 0.05). There were no major bleeds in either group, and cumulative bleeding rates by 3 months were 7.3% in the rivaroxaban group and 11.4% in the LMW heparin/vitamin K antagonist group. IMPLICATIONS: Rivaroxaban led to faster resolution of PICC-associated UEDVT than LMW/vitamin K antagonists without any increase in bleeding.

Effects of Proanthocyanidins on Intestinal Motility Disturbance Following Intestinal Ischemia/Reperfusion.

PURPOSE: To investigate the potential protective effects of Proanthocyanidins(PAs) on intestinal motility disturbance following intestinal ischemia/reperfusion (I/R). MATERIALS AND METHODS: Male rats were divided into four groups: Sham, I/R, I/R+PA100 and I/R+PA200. Sham group underwent laparotomy without ligation, the others were subjected to intestinal ischemia for 1 h and reperfusion 4 h. Rats in the I/R+PA100 group received PAs (100 mg/kg/d) for 5 days prior to I/R, while rats in the I/R+PA200 group received PAs (200 mg/kg/d). After reperfusion, using an electrophysiology instrument measured ileal slow wave. Ileal specimens were obtained to determine contractility, tissue levels of Bax, Bcl-2, and Caspase-3 and evaluate histopathological changes. In addition, blood sample was obtained to determine serum superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels. RESULTS: Intestinal I/R caused severe histopathological injury including mucosal erosions, inflammatory cell infiltration, necrosis, and hemorrhage. Both PAs treatment decreased mucosal pathological impairment in comparison with the I/R group (p < .05) in light microscopic evaluations. In both PAs-treated groups, Bax and Caspase-3 expression were decreased compared to I/R group, while the Bcl-2 expression increased (p < .05), which was similarly the case for serum SOD activity demonstrated significant enhance (p < .05) and decline in MDA levels in comparison with I/R group (both p < .05). Moreover, PAs treatment was more efficient in attenuating serum MDA levels of intestinal I/R (both p < .05). And the contractile amplitude and frequency of slow wave in I/R+PA100 and I/R+PA200 groups were higher than I/R group (both p < .05). CONCLUSIONS: PAs improve intestinal motility disturbance following intestinal I/R by alleviating oxidative stress and apoptosis.

Comparative study of the efficacy of the canalith repositioning procedure versus the vertigo treatment and rehabilitation chair.

CONCLUSION: Vertigo treatment and rehabilitation chair (TRV) may be suggested as the first choice for patients with posterior canal benign paroxysmal positional vertigo (p-BPPV). OBJECTIVE: To investigate the short- and long-term treatment efficacy of the canalith repositioning procedure (CRP) versus TRV for patients with p-BPPV. METHODS: A total of 165 patients with unilateral p-BPPV were assigned to either the CRP group or the TRV group. Patients were assessed at 1 week, 4 weeks, 3 months, and 6 months after their first treatment. The numbers of treatment sessions required for successful repositioning in both groups at 4 weeks, 3 months, and 6 months were recorded. RESULTS: Treatment efficacy of patients in the TRV group was significantly better than that of patients in the CRP group 1 week after the first treatment. The number of treatment sessions needed for successful repositioning was significantly lower in the TRV group than in the CRP group at 4 weeks and 3 months after the first treatment.

[Effect of total saponin of aralia taibaiensis on proliferation of leukemia cells].

OBJECTIVE: To investigate the effect of total saponin of Aralia Taibaiensis on the leukemia cells and explore the mechanisms of early apoptosis. METHODS: The leukemia cells lines K562 and U937 were cultured in vitro and divided into 2 groups: control group and treatment group. The proliferation inhibition of leukemia cells was measured by MTT assay. K562 cells morphological changes were observed under the reverse microscope; Flow cytometry (FCM) was used to detect the influence of total saponin of Aralia Taibaiensis on changes of mitochondrial membrane potential in K562 cells. The expression of protein BCL-2 and BAX in K562 cells was investigated by immunohistochemistry. RESULTS: The total saponin of Aralia Taibaiensis had significant inhibitory effects on K562 and U937 cell lines, and the inhibitory effects on these two cells lines presented time-dependent and dose-dependent manner. The results by inverted microscope were consistent with that of the MTT assay. FCM analysis showed that total saponin of Aralia Taibaiensis could induce mitochondrial membrane potential of K562 cells. Compared with control group, the expression of BCL-2 in treatment group was down-regulated, the expression of BAX was significantly up-regulated. CONCLUSION: The total saponin of Aralia Taibaiensis can inhibit two kinds of leukemia cells lines in vitro, and the mechanism may be related to promoting the early apoptosis of K562 cells.

[Molecular mechanism of ophiopogonin B induced cellular autophagy of human cervical cancer HeLa cells].

This study is to investigate the antitumor activity of ophiopogonin B (OP-B). MTT assay, flow cytometric analysis, acridine orange staining, Lyso-Tracker Red staining and HeLa-GFP-LC3 transfect cells assay were used to detect the proliferation activity, apoptosis and autophagy of HeLa cells. The results showed that OP-B exerted potent antiproliferative activity on HeLa cells, the cell growth inhibition effect of OP-B was not due to apoptosis and OP-B could induce autophagy of HeLa cells. OP-B also induced the protein expression up-regulation of Beclin-1 and promoted LC3 I transformation LC3 II, which were representative proteins of autophagy. Furthermore, 3-MA, an inhibitor of autophagy, not only inhibited OP-B-mediated autophagy but also almost completely reversed the antiproliferative effect of OP-B, suggesting that the growth inhibition effect of OP-B was autophagy dependent. Western blotting demonstrated that OP-B inhibited the phosphorylation of Akt and its' downstream vital protein, such as mTOR and p70S6K. In addition, OP-B also induced the protein expression up-regulation of PTEN, which is a negative regulation protein for Akt/mTOR signaling pathway. However, OP-B did not affect the protein expression of total Akt. Collectively, the antitumor effects of OP-B were autophagy-dependent via repression Akt/mTOR signaling pathway. Therefore, OP-B is a prospective inhibitor of Akt/mTOR and may be used as an alternative compound to treat cervical carcinoma.

[Synergistic antitumor effects of tanshinone II A in combination with cisplatin via apoptosis in the prostate cancer cells].

Treatment with the combination of Chinese herbs and cytotoxic chemotherapies showed a higher survival rate in clinical trials. In this report, the results demonstrated that the tanshinone II A, a key component of Salvia miltiorrhiza bunge, when it is combined with the cytotoxic drug cisplatin showed synergistic antitumor effects on human prostate cancer PC3 cells and LNCaP cells in vitro. Antiproliferative effects were detected with MTT assay. Cell cycle distribution and apoptosis were detected by flow cytometer. Protein expression was detected by Western blotting. The intracellular concentration of cisplatin was detected by high performance liquid chromatography. The results demonstrated that tanshinone II A significantly enhanced the antiproliferative effects of cisplatin on human prostate cancer PC3 cells and LNCaP cells with the increase of the intracellular concentration of cisplatin. These effects were correlated with cell cycle arrested at S phase and cell apoptosis. The apoptosis might be achieved through death receptor pathway and mitochondrial pathway. Furthermore, the Bcl-2 family members were also involved in this apoptotic process. Collectively, these results indicated that the combination of tanshinone II A and cisplatin had a better treatment effect in vitro not only on androgen-dependent LNCaP cells but also on androgen-independent PC3 cells.

Synergistic antitumor effects of tanshinone II A in combination with cisplatin via apoptosis in the prostate cancer cells / 药学学报

Treatment with the combination of Chinese herbs and cytotoxic chemotherapies showed a higher survival rate in clinical trials. In this report, the results demonstrated that the tanshinone II A, a key component of Salvia miltiorrhiza bunge, when it is combined with the cytotoxic drug cisplatin showed synergistic antitumor effects on human prostate cancer PC3 cells and LNCaP cells in vitro. Antiproliferative effects were detected with MTT assay. Cell cycle distribution and apoptosis were detected by flow cytometer. Protein expression was detected by Western blotting. The intracellular concentration of cisplatin was detected by high performance liquid chromatography. The results demonstrated that tanshinone II A significantly enhanced the antiproliferative effects of cisplatin on human prostate cancer PC3 cells and LNCaP cells with the increase of the intracellular concentration of cisplatin. These effects were correlated with cell cycle arrested at S phase and cell apoptosis. The apoptosis might be achieved through death receptor pathway and mitochondrial pathway. Furthermore, the Bcl-2 family members were also involved in this apoptotic process. Collectively, these results indicated that the combination of tanshinone II A and cisplatin had a better treatment effect in vitro not only on androgen-dependent LNCaP cells but also on androgen-independent PC3 cells.

Molecular mechanism of ophiopogonin B induced cellular autophagy of human cervical cancer HeLa cells / 药学学报

This study is to investigate the antitumor activity of ophiopogonin B (OP-B). MTT assay, flow cytometric analysis, acridine orange staining, Lyso-Tracker Red staining and HeLa-GFP-LC3 transfect cells assay were used to detect the proliferation activity, apoptosis and autophagy of HeLa cells. The results showed that OP-B exerted potent antiproliferative activity on HeLa cells, the cell growth inhibition effect of OP-B was not due to apoptosis and OP-B could induce autophagy of HeLa cells. OP-B also induced the protein expression up-regulation of Beclin-1 and promoted LC3 I transformation LC3 II, which were representative proteins of autophagy. Furthermore, 3-MA, an inhibitor of autophagy, not only inhibited OP-B-mediated autophagy but also almost completely reversed the antiproliferative effect of OP-B, suggesting that the growth inhibition effect of OP-B was autophagy dependent. Western blotting demonstrated that OP-B inhibited the phosphorylation of Akt and its' downstream vital protein, such as mTOR and p70S6K. In addition, OP-B also induced the protein expression up-regulation of PTEN, which is a negative regulation protein for Akt/mTOR signaling pathway. However, OP-B did not affect the protein expression of total Akt. Collectively, the antitumor effects of OP-B were autophagy-dependent via repression Akt/mTOR signaling pathway. Therefore, OP-B is a prospective inhibitor of Akt/mTOR and may be used as an alternative compound to treat cervical carcinoma.

Preparation and adsorption performance of 5-azacytosine-functionalized hydrothermal carbon for selective solid-phase extraction of uranium.

A new solid-phase extraction adsorbent was prepared by employing a two-step "grafting from" approach to anchor a multidentate N-donor ligand, 5-azacytosine onto hydrothermal carbon (HTC) microspheres for highly selective separation of U(VI) from multi-ion system. Fourier-transform infrared and X-ray photoelectron spectroscopies were used to analyze the chemical structure and properties of resultant HTC-based materials. The adsorption behavior of U(VI) onto the adsorbent was investigated as functions of pH, contact time, ionic strength, temperature, and initial U(VI) concentration using batch adsorption experiments. The U(VI) adsorption was of pH dependent. The adsorption achieved equilibrium within 30 min and followed a pseudo-second-order equation. The adsorption amount of U(VI) increased with raising the temperature from 283.15 to 333.15K. Remarkably, high ionic strength up to 5.0 mol L(-1) NaNO(3) had only slight effect on the adsorption. The maximum U(VI) adsorption capacity reached 408.36 mg g(-1) at 333.15K and pH 4.5. Results from batch experiments in a simulated nuclear industrial effluent, containing 13 co-existing cations including uranyl ion, showed a high adsorption capacity and selectivity of the adsorbent for uranium (0.63 mmol U g(-1), accounting for about 67% of the total adsorption amount).

Selective solid-phase extraction of uranium by salicylideneimine-functionalized hydrothermal carbon.

A new salicylideneimine-functionalized hydrothermal-carbon-based solid-phase extractant was developed for the purpose of separating uranium selectively for sustainability of uranium resources. The resulting adsorption material was obtained via hydrothermal carbonization, calcination at mild temperature (573.15K), amination, and grafting with salicylaldehyde in sequence. Both Fourier transform infrared spectra and elemental analysis proved the successful grafting of salicylideneimine onto hydrothermal carbon matrix. Adsorption behaviors of the extractant on uranium(VI) were investigated by varying pH values of solution, adsorbent amounts, contact times, initial metal concentrations, temperatures, and ionic strengths. An optimum adsorption capacity of 1.10 mmol g(-1) (261 mg g(-1)) for uranium(VI) was obtained at pH 4.3. The present adsorption process obeyed pseudo-second-order model and Langmuir isotherm. Thermodynamic parameters (ΔH=+8.81 kJ mol(-1), ΔS=+110 J K(-1)mol(-1), ΔG=-23.0 kJ mol(-1)) indicated the adsorption process was endothermic and spontaneous. Results from batch adsorption test in simulated nuclear industrial effluent, containing Cs(+), Sr(2+), Ba(2+), Mn(2+), Co(2+), Ni(2+), Zn(2+), La(3+), Ce(3+), Nd(3+), Sm(3+), and Gd(3+), showed the adsorbent could separate uranium(VI) from those competitive ions with high selectivity. The adsorbent might be promising for use in certain key steps in any future sustainable nuclear fuel cycle.

[Inducing differentiation effect on cells of serum from mice treated with Yang He decoction].

OBJECTIVE: To observe inducing differention effect on cells of serum from mice treated with Yang He decoction (SMY). METHODS: The viability of B16 cells was tested by lactate dehydrogenase (LDH) release assay and sulforhordamine B (SRB) assay. The differentiation of 3T3-L1 preadipocyte, HL-60, K562 and B16 cells were assessed by microscopy and Oil red O staining, NBT reduction and phagocytosis, haemoglobin (Hb) content and melanin content, respectively. RESULTS: SMY could obviously inhibit growth of B16 cells without cytotoxicity, and promote Oil red O staining in 3T3-L1 preadipocyte, enhance the NBT reduction ability and the phagocytosis of horse radish peroxidase in HL-60 cells, increase Hb content in K562 cells, and decrease melanin content in B16 cells. CONCLUSION: SMY is able to induce functional differentiation and maturation of 3T3-L1 preadipocyte and tumor cells, which implies the major mechanism of Yang He decoction in antitumor.