Effects of Buyang Huanwu Decoction on Intestinal Barrier, Intestinal Flora, and Trimethylamine Oxide in Rats with Heart Failure.

OBJECTIVE: To explore the mechanisms of Buyang Huanwu Decoction (BYHWD) modulating the gut microbiome and trimethylamine oxide (TAMO) to exert cardioprotective effects. METHODS: Ligation of the left anterior descending coronary artery was performed in rats to induce heart failure (HF). Except for the sham-operation group (n=10), 36 operation-induced models were randomized into 3 groups using a random number table (n=12 in each group): the model group, the BYHWD group (15.02 g/kg BYHWD), and the positive group (4.99 g/kg metoprolol succinate). After 4-week treatment (once daily by gavage), echocardiography was applied to evaluate the cardiac function and the Tei index (the ratio of ventricular isovolumic contraction time (IVCT) and isovolumic diastolic time (IVRT) to ejection time (ET)) was calculated; hematoxylin-eosin (HE) staining was observed to characterize the pathology of the myocardium and small intestinal villi. D-lactic acid was detected by an enzyme-linked immunosorbent assay (ELISA). Expressions of occludin, claudin-1, and zonula occludens (ZO-1) were detected by Western blot. 16S ribosomal ribonucleic acid (16S rRNA) sequencing was used to explore the changes in the intestinal flora. TMAO was detected via liquid chromatography-tandem mass spectrometry (LC-MS/MS). RESULTS: In the echocardiography, the Tei index was considerably lower in the positive and BYHWD groups compared with the model group (P<0.05). Besides, BYHWD improved the pathology of myocardium and small intestine of HF rats and lowered the D-lactic acid content in the serum, when compared with the model group (P<0.05). BYHWD also improved the expression of occludin and claudin-1 (P<0.05); in the gut microbiota analysis, BYHWD slowed down modifications in the structure distribution of gut microbiota and regulated the diversity of intestinal flora in HF rats. The content of TMAO in the serum was significantly lowered by BYWHT compared with the model group (P<0.05). CONCLUSION: BYHWD may delay progression of HF by enhancing the intestinal barrier structure, and regulating intestinal flora and TAMO.

Hunzeylanines A-E, Five Bisindole Alkaloids Tethered with a Methylene Group from the Roots of Hunteria zeylanica.

Five novel bisindole alkaloids, hunzeylanines A-E (1-5), with an unprecedented skeleton were isolated from the roots of Hunteria zeylanica. Compounds 1-5 represent the first examples of akuammine-pleioarpamine-type bisindole alkaloids fused with a dihydropyran unit. Their structures including absolute configurations were established through comprehensive spectroscopic data analyses and computational calculation methods. The plausible biogenetic pathway of 1 was also proposed. Alkaloids 1 and 2 displayed moderate cytotoxicity toward three human cancer cell lines (MDA-MB-231, AV3, and Huh7).

Structurally Diverse Indole Alkaloids with Vasorelaxant Activity from Melodinus hemsleyanus.

Six new structurally diverse indole alkaloids, melohemsines J-M (1-4), 11-hydroxy-Δ14-vincamine (5), and 11-hydroxy-16-epi-Δ14-vincamine (6), and 15 known alkaloids were isolated from the leaves and twigs of Melodinus hemsleyanus Diels. These new compounds and their absolute configurations were determined through spectroscopic data analyses, X-ray diffraction, and computational methods. Melohemsine J (1) is the first example of a melodinus-type alkaloid possessing a 6/6/5/5/6/5 hexacyclic skeleton and containing a tetrahydrofuro[2,3-b]pyridine-2(3H)-one unit. Melohemsine K (2) is an unusual aspidosperma-type alkaloid possessing a 6/5/6/5/5 pentacyclic architecture with a contracted E ring (loss of CH2). Compounds 5-10 and 16 exhibited vasorelaxant activities with EC50 values of 0.8-3.8 µM. In addition, compound 4 displayed moderate cytotoxicity toward the tumor cell lines HepG2 and A-549 with EC50 values of 18.7 and 28.7 µM, respectively.

[Stable Nitrite Accumulation and Phosphorus Removal from High-nitrate and Municipal Wastewaters in a Combined Process of Partial Denitrification and Denitrifying Phosphorus Removal (PD-DPR)].

In this study, a novel process combining partial denitrification (PD, NO3--N→NO2--N) and denitrifying phosphorus removal (DPR) in an anaerobic-anoxic-aerobic sequencing batch reactor (SBR) was developed. By comprehensively controlling the influent C/N ratio, anaerobic drainage ratio, and anoxic duration, the nitrite accumulation and phosphorus removal performance of a system treating high-strength nitrate and municipal wastewaters was investigated. The results showed that, after 140 days, the nitrate-to-nitrite transformation ratio (NTR) was 80.1%, and PO43--P removal efficiency was 97.64%. In the anaerobic stage (180 min), glycogen-accumulating organisms (GAOs) and phosphorus-accumulating organisms (PAOs) efficiently utilized the carbon source in municipal wastewater to enhance intracellular carbon storage. In the anoxic stage (150 min), denitrifying GAOs (DGAOs) and heterotrophic denitrifying bacteria (DOHOs) carried out endogenous and exogenous short-range denitrification, respectively, to achieve stable nitrite accumulation; simultaneously, denitrifying PAOs (DPAOs) carried out denitrifying phosphorus uptake to achieve efficient phosphorus removal. In the aerobic stage (10 min), without initiating ammonia/nitrite oxidation, PAOs absorbed excessive phosphorus, which improved the phosphorus removal performance of the system. The effluent NO2--N/NH4+-N of a ratio of 1.31:1 (close to the theoretical value of ANAMMOX process, 1.32:1), with little PO43--P and COD (0.30 and 12.94 mg·L-1), meets the requirements for deep-level nitrogen removal by coupling with ANAMMOX process.

Hunterines A-C, Three Unusual Monoterpenoid Indole Alkaloids from Hunteria zeylanica.

Three new monoterpenoid indole alkaloids (MIAs), hunterines A-C (1-3), were isolated from Hunteria zeylanica. Compound 1 possesses a unique skeleton with an unprecedented azabicyclo[4.3.1]decane ring system. Compounds 2 and 3 are a pair of epimeric vobasinylindole alkaloid heterodimers. Their structures including absolute configurations were established by spectroscopic analyses, X-ray diffraction, computational calculation, and the modified Mosher's method. Plausible biogenetic pathways of 1-3 were also proposed. Alkaloid 1 showed moderate cytotoxic activity against the HepG2 cell line.

[Nitrogen and Phosphorus Removal from Low C/N Municipal Wastewater Treated by a SPNDPR System with Different Aeration and Aerobic Times].

An anaerobic (180 min)/aerobic operated sequencing batch reactor (SBR) fed with urban sewage was optimized by regulating the aeration quantity to investigate the deep-level nitrogen (N) and phosphorus (P) removal. The amount of aeration was regulated by adjusting the volume of gas per unit volume of reactor passed in unit time, when the unit is L·(min·L)-1, from 0.125 L·(min·L)-1 gradually to 0.025 L·(min·L)-1, and aerobic times from 3 h to 6 h. The experimental results show that the effluent NH4+-N, NO2--N, NO3--N, and PO43--P concentrations of the optimized SPNDPR system were 0, 8.62, 0.06, and 0.03 mg·L-1. The effluent TN concentration was about 9.22 mg·L-1, and the TN removal efficiency was up to 87.08%. When the aeration quantity was decreased from 0.125 L·(min·L)-1 to 0.100 L·(min·L)-1; then decreased to 0.075 L·(min·L)-1, the nitrification rate of the system recovered and stabilized at 0.16 mg·(L·min)-1. However, when the aeration quantity continuously decreased to 0.050 L·(min·L)-1 and then to 0.025 L·(min·L)-1, the nitrification rate decreased to 0.09 mg·(L·min)-1 and 0.06 mg·(L·min)-1. With reduction of the aeration quantity[from 0.125 L·(min·L)-1 to 0.100, 0.075, 0.050 and 0.025 L·(min·L)-1] and extension of aerobic time (from 3 h to 6 h), the TN removal efficiency increased gradually from 62.82% to 87.08%, and the SND efficiency increased from 19.57% to 72.11%. It was proven that reducing the aeration quantity can enhance the SPND function and deep denitrification by the system was realized. By enhancing the anaerobic intracellular carbon storage and aerobic phosphorus uptake, denitrifying phosphorus removal, partial nitrification, and endogenous nitrification were achieved. The SPNDPR system, after reducing aeration and prolonging aerobic time, was able to realize deep-level denitrification and dephosphorization using low C/N urban sewage.

[Effect of the Influent C/P Ratio on the Nutrient Removal Characteristics of the SNEDPR System].

This study focuses on the nitrogen (N) and phosphorus (P) removal characteristics in a simultaneous nitrification-endogenous denitrification and phosphorus removal (SNEDPR) system at different influent C/P ratios. An extended anaerobic/low aerobic (dissolved oxygen:0.5-1.0 mg·L-1) sequencing batch reactor (SBR) fed with municipal sewage was studied by adjusting different C/P ratios (10, 15, 20, 30, and 60). The experimental results show that the proper reduction of the influent C/P ratio (C/P ratio reduced from 60 to 30) enhances the competitive advantages of phosphorus-accumulating organisms (PAOs) in the SNEDPR system. The highest phosphorus removal efficiency was achieved at a C/P ratio of 30, with the anaerobic phosphorus release rate (PRR) and aerobic phosphorus uptake rate (PUR, used as P/MLSS) reaching 3.5 mg·(g·h)-1 and 4.2 mg·(g·h)-1 respectively, and an average effluent PO43--P concentration below 0.3 mg·L-1. The percentage of PAOs contributing to the storage of endogenesis carbon (PPAO, An) reached 88.1%. However, a poor phosphorus removal performance was observed with further reduction of the influent C/P ratios to 10; both the PO43--P removal efficiency and PPAO, An decreased from 38.1% and 82.4% to 3.1% and 5.3%, respectively. The PRR and PUR were 0.2 mg·(g·h)-1 and 0.24 mg·(g·h)-1, respectively. The COD removal performance was not affected by the decreasing influent C/P ratios; the average COD removal efficiency stabilized at 85%. In addition, the nitrification performance became worse with decreasing C/P ratios (from 60 to 20) because the effluent NH4+-N and NO2--N concentrations increased from 0 and 6.9 mg·L-1 to 5.1 mg·L-1 and 16.2 mg·L-1, respectively. The nitrificaton performance recovered when the C/P ratios further decreased to 10, but the nitrite accumulation was disturbed as both the effluent NH4+-N and NO2--N concentrations reduced to 0. The effluent NO3--N concentration increased from 0.08 mg·L-1 to 14.1 mg·L-1. The SNED efficiency first decreased from 62.1% to 36.4% and then increased to 56.4%. The advantageous competition of glycogen accumulating organisms (GAOs) improved when the influent C/P ratio was lower than 15. The enhancement of the endogenous denitrification ability of GAOs might explain the recovery denitrification performance of the system when the influent C/P ratios decreased from 20 to 10.

[Effect of Different Sludge Retention Time (SRT) Operations on the Nutrient Removal Characteristics of a SNEDPR System].

This study focuses on the nitrogen (N) and phosphorus (P) removal characteristics in a simultaneous nitrification endogenous denitrification and phosphorus removal (SNEDPR) system operating at different sludge retention time (SRT). Four extended anaerobic/low aerobic (dissolved oxygen:0.5-1.0 mg·L-1)-operated sequencing batch reactors (SBRs) fed with municipal sewage were studied at different SRT of 5, 10, 15, and 25 d. The experimental results show that a shorter SRT at an SRT ≥ 10 d enhances the competitive advantage of PAOs in the system and an efficient phosphorus removal performance of the SNEDPR system was achieved at a SRT of 10 d and 15 d. Especially at an SRT of 10 d; the average PPAOs, An was 68.4%, the PRA and PUA reached 31.9 and 34.3 mg·L-1, respectively. The nitrification performance of the system was not affected by SRT changes. The most efficient nitrogen removal performance was achieved at a SRT of 15 d, with a high average TN removal and SNED efficiencies reaching 89.6% and 71.8%, respectively. At a SRT ≥ 10 d, the COD removal performance of the SNEDPR system was also not affected by SRT changes. The COD removal efficiencies were higher than 78%. However, when the SRT was shortened to 5 d, the C, N, and P performances of the system worsened due to the loss of biomass; the SNED and PO43--P removal efficiencies were as low as 5.7% and 0.5%, respectively. In addition, at an SRT=15 d, the sludge-settling performance of the system was the best. The SV and SVI were 20% and 64 mL·g-1, respectively, and the sludge concentration increased with the extension of the SRT. Under long SRT (25 d) operation, the system showed a good resistance to shock loads, but the sedimentation performance of the sludge deteriorated.

[Denitrification and Phosphorus Removal from Low C/N Urban Sewage Based on a Post-Partial Denitrification AOA-SBR Process].

This study focuses on the investigation of the nitrogen (N) and phosphorus (P) removal characteristics of a combination of enhanced phosphorus removal (EBPR) with simultaneous partial nitrification endogenous denitrification (SPND) and post-partial denitrification process. An anaerobic/aerobic/anoxic (A/O/A) operated sequencing batch reactor (SBR) fed with urban sewage was optimized by regulating the aeration rate and anoxic time. Based on this optimization, deep-level nitrogen and phosphorus removals from low C/N urban sewage could be realized. The experimental results show that the effluent PO43--P concentration decreased from 0.06 mg·L-1 to 0 mg·L-1, the effluent NH4+-N, NO2--N, and NO3--N concentrations gradually decreased from 0.18, 18.79, and 0.08 mg·L-1 to 0, 16.46, and 0.05 mg·L-1, respectively, and the TN removal efficiency increased from 72.69% to 77.97% when the aeration rate decreased from 1.0 L·min-1 to 0.6 L·min-1 and the anoxic duration was 180 min. With the reduction of the aeration rate, the SPND phenomenon became notable and the SND rate increased from 19.18% to 31.20%. When the anoxic duration was extended from 180 min to 420 min, the effluent PO43--P, NH4+-N, and NO3--N concentrations stabilized at~0, 0, and 0.03 mg·L-1, respectively. The effluent NO2--N concentration was as low as 3.06 mg·L-1, the SND rate was~32.21%, the TN removal performance gradually improved, and the TN removal efficiency was as high as 99.42%. Thus, deep-level nitrogen and phosphorus removals could be realized with the SPNDPR-PD system.

[Operating Characteristics of a DPR-SNED System Treating Low C/N Municipal Wastewater and Nitrate-containing Sewage].

In order to realize the simultaneous treatment of low C/N municipal wastewater and high nitrate wastewater, a sequencing batch reactor (SBR), inoculated with activated sludge, was used to initiate the denitrifying phosphorus removal coupled with simultaneous nitrification and endogenous denitrification (DPR-SNED). The anaerobic/anoxic/hypoxic durations and dissolved oxygen (DO) concentration were appropriately controlled, and the nitrogen and phosphorus removal characteristics were examined. The experimental results demonstrated that, in the anaerobic/hypoxia operation mode, with an anaerobic duration of 3 h and DO concentration of 0.5-1.0 mg·L-1, the simultaneous nitrification of phosphorus removal (SNEDPR) system successfully began in 60 d. The effluent PO43--P concentration was below 0.5 mg·L-1, the nutrient and COD removal efficiencies were stably maintained above 90% and 80%, respectively, and the SNED efficiency and CODins efficiency reached 70% and 95%, respectively. When the operation mode was anaerobic/anoxic/hypoxic and nitrate-containing sewage was added at the beginning of the anoxic stage, DPR-SNED was achieved with the effluent PO43--P concentration<0.5 mg·L-1, nutrient and COD removal efficiencies above 88% and 90%, respectively, and SNED efficiency and CODins efficiency maintained at 62% and 90%, respectively. After the successful initiation of DPR-SNED, enhanced intracellular carbons storage was achieved by phosphorus-and glycogen-accumulating organisms using the limited carbons in raw municipal wastewater to provide sufficient carbon sources for subsequent nutrient removal. In addition, the endogenous partial denitrification ensured the efficient nitrogen removal performance of the DPR-SNED system at low C/N conditions (average 4).

[Simultaneous Nitrogen and Phosphorus Removal Characteristics of An Anaerobic/Aerobic Operated SPNDPR System Treating Low C/N Urban Sewage].

This study focused on the nitrogen (N) and phosphorus (P) removal performance optimization of simultaneous partial nitrification-endogenous denitrification and phosphorus removal (SPNDPR) systems. An anaerobic (180 min)/aerobic operated sequencing batch reactor (SBR) fed with domestic wastewater was used for investigating the startup and optimization of SPNDPR by regulating the aeration rate and aerobic duration time. The experimental results showed that at an aerobic aeration rate of 0.8 L·min-1 and aerobic duration time of 150 min, the effluent PO43--P concentration was about 1.5 mg·L-1, with the effluent NH4+-N and NO3--N concentrations gradually decreasing from 10.28 and 8.14 mg·L-1 to 0 and 2.27 mg·L-1, respectively, and effluent NO2--N concentration increasing to 1.81 mg·L-1. When the aeration rate was increased to 1.0 L·min-1 and the aerobic duration time was shortened to 120 min, the phosphorus removal and partial nitrification-endogenous performance of the system gradually increased, but the total nitrogen (TN) removal performance initially decreased and then gradually increased. The final effluent PO43--P and NH4+-N were stably below 0.5 and 1.0 mg·L-1, respectively, aerobic nitrite accumulation and simultaneous nitrification-endogenous denitrification (SND) efficiencies were 98.65 and 44.20%, respectively, and TN removal efficiency was 79.78%. The concurrence of aerobic phosphorus absorption, denitrifying phosphorus removal, partial nitrification, and nitrification-endogenous in the aerobic stage of the SPNDPR system ensured the simultaneous removal of N and P from low C/N wastewater.