Traditional Chinese medicine CFF-1 exerts a potent anti-tumor immunity to hinder tumor growth and metastasis in prostate cancer through EGFR/JAK1/STAT3 pathway to inhibit PD-1/PD-L1 checkpoint signaling.

BACKGROUND: Traditional Chinese Medicine (TCM) CFF-1 has been used in clinic for prostate cancer therapy in China. We reported before CFF-1 induced cell apoptosis via suppressing EGFR-related pathways, reminding us its potential role associated with antitumor immunity. PURPOSE: The study was aimed to investigate the regulatory mechanism of CFF-1 on PD-L1/PD-1-mediated tumor immune escape. METHODS: Prostate-specific antigen (PSA) test and the functional assessment of cancer therapy-prostate (FACT-P) and karnosky performance status (KPS) questionnaires were carried out to evaluate patient' condition before and after therapy. Flow cytometry (FCM) was used for analyzing cell apoptosis, T lymphocyte subsets and cell cycle. Western blotting and Immunohistochemistry (IHC) were performed to measure protein expressions. The synergy of drug combination was assessed by calculating combination index (CI). RESULTS: CFF-1 obviously decreased PSA and improved the quality of life in patients with advanced prostate cancer. PD-L1 was highly expressed in prostate cancer cells including LNCaP, 22Rv1, PC-3, DU145 and RM-1. PD-1/PD-L1 was upregulated in tumorigenesis and tumor progression of subcutaneous homograft mouse model with immune response, where CD3+ T cell subsets were declined. CFF-1 inhibited PD-L1 expression in prostate cancer cells in a time/dose-dependent manner and blocked tumor growth by suppressing PD-1/PD-L1 upregulation to promote the recovery of CD3+ T lymphocytes, especially CD4+ T cell subset, accompanied by the downregulation of CD4+ FOXP3+ T cell subset. CFF-1 also prolonged the survival and inhibited lung metastasis in tail vein prostate cancer mouse model while repressing PD-1/PD-L1. CFF-1 in combination with docetaxol (DTX) produced a synergistic effects by sensitizing the inhibitory effect of DTX on JAK1/STAT3 pathway targeting PD-L1 blockade. CONCLUSION: CFF-1 inhibited tumor growth and lung metastasis by blocking PD-1/PD-L1 to ameliorate T lymphocyte immune response through EGFR/JAK1/STAT3 pathway, suggesting that CFF-1 might be a promising treatment to resist tumor immunosuppression for prostate cancer patients.

Carbon fluxes response of an artificial sand-binding vegetation system to rainfall variation during the growing season in the Tengger Desert.

Revegetation is considered as an effective approach for desertification control, and artificial sand-binding vegetation exerts a significant contributor to carbon cycling in arid and semiarid regions; however, this is largely determined by the rainfall regime. We measured carbon fluxes (the net ecosystem CO2 exchange (NEE), the gross ecosystem productivity (GEP) and the ecosystem respiration (Reco)) during the growing season of 2014-2016 using the eddy covariance technique and explored the effects of rainfall variables (amount, timing distribution and pulse size) and environmental factors on carbon fluxes at different time scales. The system had NEE values of -117.5 and -98.9 g C m-2 during the growing seasons of 2015 (dry year) and 2016 (wet year), respectively. When the rainfall amount did not differ significantly between spring and autumn, the cumulative GEP was greater in spring than in autumn, whereas the cumulative Reco and NEE showed the opposite pattern. Small (<5 mm) rain events failed to trigger obvious GEP and NEE pulses, whereas ≥ 5 mm or a series of small rain events led to high assimilation but with hysteresis. The magnitude of Reco increased as the rain pulses increased. The Random Forest (RF) algorithm revealed that soil water contents had a great impact on carbon fluxes at different integration periods. A correlation analysis showed that the soil water contents were positively correlated with GEP and Reco and negatively correlated with NEE over different time scales in most cases. These findings suggest that artificial vegetation not only improves habitat restoration but is a significant carbon sink during both dry and wet growing season, which is likely to supplement our knowledge gap to accurately evaluate the current carbon budget in dry land.

EDTA-Modified 17ß-Estradiol-Laden Upconversion Nanocomposite for Bone-Targeted Hormone Replacement Therapy for Osteoporosis.

Hormone therapy (HT) is one of the most effective treatments for osteoporosis. However, the nonselective accumulation of hormone in organs such as breast, heart and uterus other than bones causes serious side effects, which impedes the application of HT. Hence, it is critically important to develop a HT strategy with reduced non-specific enrichment of hormone drugs in non-target tissues and enhanced bone-targeting ability. Methods: Herein, a 17ß-estradiol (E2)-laden mesoporous silica-coated upconversion nanoparticle with a surface modification of ethylenediaminetetraacetic acid (EDTA) (NaLuF4:Yb,Tm@NaLuF4@mSiO2-EDTA-E2, E2-csUCNP@MSN-EDTA) is developed for bone-targeted osteoporosis hormone therapy. EDTA was attached onto the surface of E2 upconversion nanocomposite to enhance its affinity and efficiency targeting bone tissue and cells to optimize hormone replacement therapy for osteoporosis. We characterized the size, cytotoxicity, loading and release efficiency, in situ and ex vivo imaging. Further, in vitro and in vivo osteogenic ability was tested using preosteoblast and ovariectomy mouse model of osteoporosis. Results: The upconversion core of E2-csUCNP@MSN-EDTA nanoparticle serves as an excellent imaging agent for tracking the loaded hormone drug in vivo. The mesoporous silica layer has a high loading efficiency for E2 and provides a relatively long-lasting drug release within 50 h. EDTA anchored on the silica layer endows the nanocomposite with a bone targeting property. The nanocomposite effectively reverses estrogen deficiency-induced osteoporosis and reduces the damage of hormone to the uterus. The bone mineral density in the nanocomposite treatment group is nearly twice that of the ovariectomized (OVX) group. Compared with the E2 group, the uterine weight and luminal epithelial height were significantly lower in the nanocomposite treatment group. Conclusion: This work demonstrated that E2-csUCNP@MSN-EDTA alleviates the side effect of hormone therapy while maintaining its therapeutic efficacy, which has great potential for developing the next generation of methods for osteoporosis treatment.

L-Theanine Down-Regulates the JAK/STAT3 Pathway to Attenuate the Proliferation and Migration of Vascular Smooth Muscle Cells Induced by Angiotensin II.

L-Theanine, a green tea amino acid derivative, has cardiovascular qualities. The focus of the current evaluation was to examine the suppression of L-theanine on cultured vascular smooth muscle cell (VSMC) proliferation and migration that is prompted by angiotensin II (Ang II). The VSMCs were treated with non-cytotoxic concentrations of L-theanine and then stimulated with Ang II. The CCK-8 and Transwell chamber assays were monitored on the proliferation and migration rate, respectively. We discovered that L-theanine (50 and 100 µM) significantly halted Ang II-induced VSMC proliferation and migration. This was joined by a decline in the amount of cyclin D1. An additional discovery was that L-theanine lowered the proportion of S-phase cells, whereas the number of G1/G0-phase cells in Ang II-stimulated VSMCs was elevated, based on flow cytometry. Western blotting analyses indicated that L-theanine had no impact on extracellular-signal-regulated kinase 1/2 (ERK1/2) activation prompted by Ang II. Nevertheless, L-theanine significantly lowered Ang II-prompted phosphorylation of Janus kinase 2 (JAK2), c-Src tyrosine kinase, and signal transducer and activators of transcription 3 (STAT3). The outcomes revealed that L-theanine subdued the Ang II-prompted proliferation and migration of VSMC, partly via the obstruction of the JAK/STAT3 pathway instead of via just the ERK pathway.

[Acupuncture with smoothing liver and regulating qi for post-stroke slow transit constipation and its gastrointestinal hormone level].

OBJECTIVE: To compare the efficacy between acupuncture with smoothing liver and regulating qi and lactulose for post-stroke slow transit constipation(STC) and to explore the mechanism. METHODS: Sixty patients were randomized into an acupuncture group and a medication group,30 cases in each one. Based on the comprehensive stroke unit care,acupuncture with smoothing liver and regulating qi was used at Danzhong(CV 17),Qihai(CV 6),Tianshu(ST 25),Neiguan(PC 6),Gongsun(SP 4) and Taichong(LR 3) in the acupuncture group,once a day. Lactulose oral liquid was taken at a draught in the morning in the medication group,20 to 30 mL a time,once a day. The study period was 11 weeks,including 1-week baseline evaluation,6-week treatment and 4-week follow-up. We recorded the time of the first independent defecation,constipation symptom score,and gastrointestinal hormone level,including somatostatin(SS),motilin(MTL),P substance(SP) and vasoactive intestinal peptide(VIP). Also,the side effects were recorded at any time. RESULTS: The time of the first independent defecation was (30.18±16.14) h in the acupuncture group,which was significantly different from (43.22±28.42) h in the medication group(P<0.05). The constipation scores after 6-week treatment and at follow-up were lower than those before treatment in the two groups (all P<0.05),with better results in the acupuncture group(both P<0.05). MTL and SP increased,as well as SS and VIP decreased after treatment in the two groups(all P<0.05). The changes were better in the acupuncture group(all P<0.05). The side effect was not observed in the two groups. CONCLUSIONS: Acupuncture with smoothing liver and regulating qi achieves better effect than lactulose for post-stroke STC in terms of efficacy onset,extent,and long term. The mechanism may relate to increasing excitatory regulatory peptide and reducing inhibitory regulatory peptide.

Emodin promotes the osteogenesis of MC3T3-E1 cells via BMP-9/Smad pathway and exerts a preventive effect in ovariectomized rats.

Emodin, a natural anthraquinone extracted from the Chinese herbs rhubarb and giant knotweed rhizome, has been reported to enhance osteoblast differentiation. However, the mechanisms underlying its ability to regulate osteogenesis are unclear. The objective of this study was to determine the role of emodin in osteoblast function in vitro and its osteoprotective effect in vivo. Emodin enhanced the differentiation and mineralization of MC3T3-E1 cells, as evidenced by elevated alkaline phosphatase activity and increased number of mineralized nodules. In cultured osteoblasts, emodin significantly induced the mRNA expression of BMP-9 which is one of the least studied but most osteogenic bone morphogenetic proteins (BMPs). Furthermore, the bone morphogenetic protein receptor-Smad (BMPR-Smad) signaling axis and p38 mitogen activated protein kinase (p38 MAPK) were activated. The in vivo function of emodin were evaluated by assessing bone histomorphology, trabecular bone microarchitecture, mechanical properties of the skeleton, and serum parameters of bone turnover in ovariectomized (OVX) rats. Emodin combined with low-dose of estrogen improved trabecular bone microarchitecture in the fourth lumbar vertebra compared with low-dose estrogen alone and enhanced vertebral body strength. Moreover, emodin suppressed the OVX-induced elevation of serum osteocalcin (OC). In addition, there were fewer side effects on uterine hypertrophy with the combination therapy than with high-dose estrogen alone. However, emodin alone did not exert any osteoprotective effect. These results suggest that emodin may be a promising alternative agent for osteoporosis in combination therapy.

l-Theanine attenuates cadmium-induced neurotoxicity through the inhibition of oxidative damage and tau hyperphosphorylation.

Cadmium (Cd) has long been known to induce neurological degenerative disorders. We studied effects of l-theanine, one of the major amino acid components in green tea, on Cd-induced brain injury in mice. Male ICR mice were intraperitoneally injected with l-theanine (100 or 200mg/kg/day) or saline and after one hour these mice were orally administrated with CdCl2 (3.75-6mg/kg). The treatment was conducted for 8 weeks. l-Theanine significantly reduced Cd level in the mouse brain and plasma. Cd-induced neuronal cell death in the mouse cortex and hippocampus were apparently inhibited by l-theanine treatment. l-Theanine also decreased the levels of malondialdehyde (MDA) and ROS, and obviously elevated the levels of glutathione (GSH) and activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in the mouse brain. Hyperphosphorylation of tau protein is proposed to be an early event for the evolution of tau pathology, and may play an important role in Cd-induced neurodegeneration. Our results showed that l-theanine significantly suppressed Cd-induced tau protein hyperphosphorylation at Ser199, Ser202, and Ser396. Mechanism study showed that l-theanine inhibited the activation of glycogen synthase kinase-3ß (GSK-3ß) which contributed to the hyperphosphorylation of tau and Cd-induced cytotoxicity. Furthermore, l-theanine reduced Cd-induced cytotoxicity possibly by interfering with the Akt/mTOR signaling pathway. In conclusion, our study indicated that l-theanine protected mice against Cd-induced neurotoxicity through reducing brain Cd level and relieved oxidative damage and tau hyperphosphorylation. Our foundings provide a novel insight into the potential use of l-theanine as prophylactic and therapeutic agents for Cd-induced neurodegenerative diseases.

[Effect of Modified Hangqi Chifeng Decoction Containing Serum on the Expression of Col IV, MMP-2, and TIMP-2 in Glomerular Mesangial Cells Induced by LPS].

OBJECTIVE: To explore the effect of Modified Hangqi Chifeng Decoction (MHCD) on levels of collagen type IV (Col IV), matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-2 (TIMP-2) in extracellular matrix (ECM) of glomerular mesangial cells (GMCs) in LPS induced mice. METHODS: Normal serum and telmisartan, high, medium, low dose MHCD containing serums were prepared by using serum pharmacology method. GMCs were cultured in vitro. The proliferation of mesangial cells were induced using LPS as stimulating factor. GMCs were divided into six groups, i.e., the normal group, the model group, the telmisartan group, high, medium and low dose MHCD groups. Col IV content in the supernatant of mesangial cells was detected using ELISA. Protein expressions of MMP-2 and TIMP-2 were detected using Western blot. RESULTS: Compared with the normal group, Col IV content obviously increased in the model group after 72-h LPS stimulation; protein expressions of MMP-2 and TIMP-2 were obviously up-regulated, and MMP-2/TIMP-2 ratio was down-regulated in the model group (P < 0.01). Compared with the model group, Col IV content obviously decreased in high and medium dose MHCD groups and the telmisartan group (P < 0.01); protein expressions of MMP-2 were obviously down-regulated in medium and low dose MHCD groups (P < 0.01, P < 0.05); the protein expression of TIMP-2 was obviously down-regulated in high, medium, low dose MHCD groups and the telmisartan group (P < 0.01). The pro- tein expression of TIMP-2 was obviously lower in the high dose MHCD group than in the low dose MHCD group (P < 0.01). MMP-2/TIMP-2 ratio was obviously up-regulated in the telmisartan group, high and medium dose MHCD groups (P < 0.01). CONCLUSION: MHCD could regulate disordered MMP-2/TIMP-2 ratio in LPS induced ECM, inhibit excessive production of Col IV in ECM, promote the degradation of ECM, reduce the accumulation of ECM, thereby, delaying the process of glomerular sclerosis.

[Anti-renal Fibrosis Mechanism of Modified Huangqi Chifeng Decoction Based on TGF-p1/Smad Signal Pathway].

Objective To observe the effect of Modified Huangqi Chifeng Decoction (MHCD) on TGF-ß1/Smad signal pathway, and to explore its anti-renal fibrosis mechanism. Methods Adopting ser- um pharmacology method, rats were intragastrically administered with MHCD and telmisartan to prepare drug containing serum. Mouse mesangial cells were cultured in vitro, using lipopolysaccharides (LPS) as stimulating factor. The cells were divided into six groups, i.e., the normal group, the model group, the telmisartan group, high, medium and low dose MHCD groups. The cell supernatant was collected in each group after mouse mesangial cells were intervened by drug containing serum for 72 h. Contents of laminin (LN) and fibronectin (FN) were measured using ELISA. Protein expression levels of TGF-P, , p-Smad2/3, Smad7, and connective tissue growth factor (CTGF) were detected using Western blot. Results (1) Compared with the normal group, contents of LN and FN in supernatant significantly increased in the model group (P <0. 01). Compared with the model group, contents of LN and FN were significantly reduced in the telmisartan group and the medium dose MHCD group (P <0. 05, P <0. 01). FN content in su- pernatant significantly decreased in the low dose MHCD group (P <0. 01). (2) Compared with the normal group, protein expressions of TGF-ß1 , p-Smad2/3, and CTGF were significantly increased, Smad7 protein expression significantly decreased in the model group, with statistical difference (P <0. 01). Compared with the model group, protein expressions of TGF-ß1 and CTGF significantly decreased in the telmisartan group and 3 MHCD groups (P <0. 01 , P <0. 05) ; protein expression of p-Smad23 significantly decreased in the telmisartan group, high and medium dose MHCD groups (P <0.01); Smad7 protein expression were significantly increased in high and medium dose MHCD groups (P <0. 05). Conclusion MHCD could inhibit increased inflammatory factors induced secretion of extracellular matrix in glomerular mesangial cells, and restrain excessive activation of TGF-ß1/Smad signal pathways, which might be one of its anti-renal fibrosis mechanisms.

Correlations between serum intact parathyroid hormone (PTH) and N-terminal-probrain natriuretic peptide levels in elderly patients with chronic heart failure (CHF).

OBJECTIVES: The aim of this study was to investigate the association between PTH and Nt-proBNP in elderly patients with CHF in an attempt to gain insights into the role of PTH in a community-based cohort of elderly patients with CHF. METHODS: A total of 182 consecutive CHF patients with follow-up for mortality after 3 years were prospectively studied. Serum levels of intact PTH, Nt-proBNP and biochemical parameters were examined. The enrolled patients were divided into groups by the levels of PTH and New York Heart Association (NYHA) functional classes. RESULTS: A total of 66 (36%) patients had PTH values above the upper limit of the normal range. Serum creatinine (p=0.001), estimated glomerular filtration rate (eGFR) (p=0.001), Nt-proBNP (p<0.001), serum calcium (p=0.030), heart rate (p=0.002) showed statistical significance in different stages of PTH. The mean PTH and Nt-proBNP levels increased as the NYHA functional class increased. The optimal cut-off value of PTH to predict CHF-related death was 48.98 pg/ml, with 57.14% sensitivity and 86.24% specificity. The best cut-off point of Nt-proBNP was 480 ng/ml with 76.47% sensitivity and 80.48% specificity. Over a mean follow-up of 3 years, Kaplan-Meier survival curves demonstrate that patients with higher levels of intact PTH had lower survival time, with a hazard ratio of 2.5 (95% CI 1.5-3.9). CONCLUSIONS: The study has shown that serum intact PTH level obtained in the elderly patients with CHF is a novel biomarker associated with Nt-proBNP and could provide supplementary information for the diagnosis and prognostic prediction of CHF, especially when it is used in combination with Nt-proBNP.

Leaf nitrogen and phosphorus of temperate desert plants in response to climate and soil nutrient availability.

In desert ecosystems, plant growth and nutrient uptake are restricted by availability of soil nitrogen (N) and phosphorus (P). The effects of both climate and soil nutrient conditions on N and P concentrations among desert plant life forms (annual, perennial and shrub) remain unclear. We assessed leaf N and P levels of 54 desert plants and measured the corresponding soil N and P in shallow (0-10 cm), middle (10-40 cm) and deep soil layers (40-100 cm), at 52 sites in a temperate desert of northwest China. Leaf P and N:P ratios varied markedly among life forms. Leaf P was higher in annuals and perennials than in shrubs. Leaf N and P showed a negative relationship with mean annual temperature (MAT) and no relationship with mean annual precipitation (MAP), but a positive relationship with soil P. Leaf P of shrubs was positively related to soil P in the deep soil. Our study indicated that leaf N and P across the three life forms were influenced by soil P. Deep-rooted plants may enhance the availability of P in the surface soil facilitating growth of shallow-rooted life forms in this N and P limited system, but further research is warranted on this aspect.

Hollow silica nanoparticles loaded with hydrophobic phthalocyanine for near-infrared photodynamic and photothermal combination therapy.

Owing to the convenience and minimal invasiveness, phototherapy, including photodynamic therapy (PDT) and photothermal therapy (PTT), is emerging as a powerful technique for cancer treatment. To date, however, few examples of combination PDT and PTT have been reported. Phthalocyanine (Pc) is a class of traditional photosensitizer for PDT, but its bioapplication is limited by high hydrophobicity. In this present study, hollow silica nanospheres (HSNs) were employed to endow the hydrophobic phthalocyanine with water-dispersity, and the as-prepared hollow silica nanoparticles loaded with hydrophobic phthalocyanine (Pc@HSNs) exhibits highly efficient dual PDT and PTT effects. In vitro and in vivo experimental results clearly indicated that the dual phototherapeutic effect of Pc@HSNs can kill cancer cells or eradicate tumor tissues. This multifunctional nanomedicine may be useful for PTT/PDT treatment of cancer.

[Simultaneous determination of eight iridoid glycosides in Gardeniae fructus by HPLC].

OBJECTIVE: To establish an HPLC method for simultaneous determination of eight iridiods in Gardeniae Fructus. METHOD: Kromasil C18 column (4. 6 mm x 250 mm, 5 microm) was adopted, with acetonitrile-water-trifluoroacetic acid (6:94: 0.05) as the mobile phase at the flow rate of 1.0 mL x min(-1). The detection wavelength was set at 238 nm, and the column temperature was 40 degrees C. RESULT: The linear ranges of geniposide, gardoside, shanzhiside, geniposidic acid, deacetyl asperulosidic acid methyl ester, gardenoside, scandoside methyl ester, and genipin gentiobioside were 1.5036 - 15.036, 0.04256 - 0.4256, 0.1038 - 1.038, 0.00992 - 0.0992, 0.02332 - 0.2332, 0.4128 - 4.128, 0.02040 - 0.2040 and 0.4656 - 4.656 microg, respectively. Their average recoveries were 99.6% , 100.6% , 101.2%, 99.5%, 100.3% , 98.7%, 99.8% and 100.1%, respectively. CONCLUSION: The method shows good separation and it is so simple, accurate and highly repeatable that it can be used for providing basis for quality control of Gardeniae Fructus.

Hexaoxygenated flavonoids from Pteroxygonum giraldii.

Two flavonoids with an unusual 2',4',5'-trisubstituted B-ring (1 and 2), four myricetin derivatives (3-methylmyricetin-3'-O-beta-D-xylopyranoside (3), myricetin-3-O-alpha-L-rhamnopyranoside, myricetin-3-O-beta-D-galactopyranoside, and 3-methylmyricetin), and myricetin were isolated from the roots of Pteroxygonum giraldii Damm. & Diels. Their structures were elucidated using various spectroscopic methods and acid hydrolysis. Compound 1 was a new flavonoid and the NMR spectroscopic data of compounds 2 and 3 are reported for the first time.

Highly sensitive and fast responsive fluorescence turn-on chemodosimeter for Cu2+ and its application in live cell imaging.

A rhodamine B derivative 4 containing a highly electron-rich S atom has been synthesized as a fluorescence turn-on chemodosimeter for Cu(2+). Following Cu(2+)-promoted ring-opening, redox and hydrolysis reactions, comparable amplifications of absorption and fluorescence signals were observed upon addition of Cu(2+); this suggests that chemodosimeter 4 effectively avoided the fluorescence quenching caused by the paramagnetic nature of Cu(2+). Importantly, 4 can selectively recognize Cu(2+) in aqueous media in the presence of other trace metal ions in organisms (such as Fe(3+), Fe(2+), Cu(+), Zn(2+), Cr(3+), Mn(2+), Co(2+), and Ni(2+)), abundant cellular cations (such as Na(+), K(+), Mg(2+), and Ca(2+)), and the prevalent toxic metal ions in the environment (such as Pb(2+) and Cd(2+)) with high sensitivity (detection limit < or =10 ppb) and a rapid response time (< or =1 min). Moreover, by virtue of the chemodosimeter as fluorescent probe for Cu(2+), confocal and two-photon microscopy experiments revealed a significant increase of intracellular Cu(2+) concentration and the subcellular distribution of Cu(2+), which was internalized into the living HeLa cells upon incubation in growth medium supplemented with 50 muM CuCl(2) for 20 h.