Biocontrol of strawberry Botrytis gray mold and prolong the fruit shelf-life by fumigant Trichoderma spp.

Objectives To screen high active volatile organic compounds (VOCs)-producing Trichoderma isolates against strawberry gray mold caused by Botrytis cinerea, and to explore their antagonistic mode of action against the pathogen. VOCs produced by nine Trichoderma isolates (Trichoderma atroviride T1 and T3; Trichoderma harzianum T2, T4 and T5; T6, T7, T8 and T9 identified as Trichoderma asperellum in this work) significantly inhibited the mycelial growth (13.9-63.0% reduction) and conidial germination (17.6-96.3% reduction) of B. cinerea, the highest inhibition percentage belonged to VOCs of T7; in a closed space, VOCs of T7 shared 76.9% and 100% biocontrol efficacy against gray mold on strawberry fruits and detached leaves, respectively, prolonged the fruit shelf-life by 3 days in presence of B. cinerea, completely protected the leaves from B. cinerea infecting; volatile metabolites of T7 damaged the cell membrane permeability and integrity of B. cinerea, thereby inhibiting the mycelial growth and conidial germination. Gas chromatography-mass spectrometry (GC-MS) analysis revealed the VOCs contain 23 potential compounds, and the majority of these compounds were categorised as alkenes, alcohols, and esters, including PEA and 6PP, which have been reported as substances produced by Trichoderma spp. T. asperellum T7 showed high biofumigant activity against mycelial growth especially conidial germination of B. cinerea and thus protected strawberry fruits and leaves from gray mold, which acted by damaging the pathogen's plasma membrane and resulting in cytoplasm leakage, was a potential biofumigant for controlling pre- and post-harvest strawberry gray mold.

[The prognostic evaluation of coronary artery calcification score and CT fractional flow reserve for patients with stable coronary artery disease].

Objective: To investigate the prognostic value of coronary artery calcium score (CACS) and computed tomography-derived fractional flow reserve (CT-FFR) for major adverse cardiovascular events (MACE) in patients with stable coronary artery disease (CAD). Methods: The data for this prospective study were derived from a prospective clinical trial at a single center. This trial enrolled stable CAD patients who underwent coronary CT angiography (CCTA) in General Hospital of Eastern Theater Command from April 2018 to March 2019 and had coronary artery stenosis of 25%-80%. Patients were assigned to either the control group or trial group according to CCTA time. Patients in control group were provided with only a standard CCTA report, while patients in trial group were provided with both a standard CCTA report and the corresponding CT-FFR results. The study included patients who underwent ECG-gated calcium scoring CT scans in this trial. CT-FFR value at 2 cm distal to the narrowest stenosis of each vessel was calculated. The minimum CT-FFR value was recorded as the patient level and CT-FFR≤0.80 was defined as a positive result. All patients were followed up for MACE, including all-cause death, nonfatal myocardial infarction, and acute coronary syndrome leading to unplanned revascularization. Multivariable Cox proportional hazards regression analysis was used to identify variables associated with MACE occurrence, and the Concordance index (C-index) was used to represent the performance of the models for predicting MACE occurrence based on clinical, anatomical, and CT-FFR parameters. Results: A total of 783 patients were finally statistically analyzed, with a age of (62.0±10.8) years, of whom 64.6% (506 cases) were male. There were 383 patients in the trial group and 400 patients in the control group, with a median follow-up time of 35.3 months. A total of 81 MACE cases occurred during the follow-up. The incidence of MACE in trial group (8.1%, 31/383) was significantly lower than that in control group (12.5%, 50/400)(χ2=4.095, P=0.043). CACS≥300, stenosis≥70% and CT-FFR≤0.80 [HR (95%CI) were 2.14 (1.01-4.52), 5.38 (3.44-8.42) and 16.91 (9.21-31.04), all P<0.05] showed predictive value for MACE. The predictive ability of the CT-FFR model is significantly better than that of the CACS model and the stenosis degree model [C-index (95%CI) were 0.850 (0.823-0.874), 0.653 (0.618-0.686) and 0.718 (0.685-0.749), all P≤0.001]. The comprehensive model with added CACS and stenosis degree did not significantly improve the predictive value of the CT-FFR model [C-index (95%CI) were 0.867 (0.841-0.890), 0.850 (0.823-0.874), P=0.584]. Conclusions: CT-FFR has a high predictive value for MACE in patients with stable CAD, the combination of CT-FFR and CACS did not increase the predictive power of CT-FFR.

[Simultaneous combined radical surgery for hepatic and renal alveolar echinococcosis: an analysis of 10 clinical cases].

Objective: To examine the feasibility, safety and efficacy of simultaneous combined radical surgery for hepatic and renal alveolar echinococcosis (AE). Methods: Clinical dates of consecutive 10 hepatic and renal AE patients who accepted surgical treatment in the First Affiliated Hospital of Xinjiang Medical University during April 2013 to September 2019 were analyzed retrospectively. There were 8 males and 2 females, aged (42.5±10.3) years (range: 27 to 52 years). Seven of them had previously palliative surgical treatment with poor compliance to post-operative medication. All of the patients had hepatic-renal combined AE lesions, and two of them had left lateral and left renal lesions for each, which sized for (726.4±576.1)cm3 (range: 117.0 to 1 998.0 cm3). Extra-hepatic or renal lesions infiltrated to right diaphragm, inferior vena cava, right adrenal gland, abdominal wall, right psoas muscle, duodenum wall and right pulmonary lobe (respectively were 7, 7, 6, 2, 1, 1, 2 cases). Surgery were performed mainly with simultaneous combined surgery and vascular reconstruction techniques for this series. Hemi-hepatectomy or extended right lobectomy was applied in 8 patients, while 2 other patients received ex vivo liver resection and autotransplantation, additionally, one patient had partial hepatectomy for left lateral lobular lesion. Total right nephrectomy, partial right nephrectomy and partial left nephrectomy were respectively performed on 7, 3 and 1 patient(s). Additionally, extra-hepatic or renal lesions were eradicated followed by relevant repairments or reconstructions. Results: Surgeries went well and there was no intra-or post-operative liver or renal dysfunction occurred. During recovery period, 3 cases experienced with hydrothorax and managed well after drainage and supportive treatment, and one patient developed peri-renal urinary leakage and cured by "J" catheter. The subjects were followed-up for 6 to 81 months (median: 21 months), no death, organ dysfunction, chronic or acute kidney diseases occurred. One case encountered with abdominal hernia at post-operative 7th month and was successfully managed with laparoscopic repair with artificial mesh. No disease recurrence in all patients, which reached clinical cure at last. Conclusion: When complied strictly to indications, simultaneous combined radical surgery could be a feasible, safe and efficient approach for patients with hepatic and renal AE, which is primary or relapsed from previous hepatic AE surgery or interventional therapies as well as neighboring organ AE invasions.

[The clinical practice of improvement the "Volume and Quality" of functional liver in autologous liver transplantation for the patients with alveolar echinococcosis].

Objective: To investigate the clinical significance of accurate assessment of "volume and quality" of functional liver in Autologous liver transplantation (ALT) in the treatment of the advanced hepatic alveolar echinococcosis (HAE). Methods: The clinical data of 12 patients with advanced HAE who underwent ALT at the First Affiliated Hospital of Xinjiang Medical University from May 2015 to July 2016 were retrospectively analyzed. Results: The preoperative hepatic functions of 12 patients were 8 Child-Pugh Grade A, 1 Grade B, and 3 Grade C. Three of the patients had moderate or severe jaundice. Three of the patients calculated functional liver graft volume (GV) and standard liver volume (SLV) ratio (GV/SLV) were <30%. After the protection of liver function, anti-infection, percutaneous transhepatic cholangiography drainage (PTCD), selective portal vein embolization (PVE), and staging liver resection, liver function Child-Pugh grade of 11 patients was raised to A grade, and the other patient was B grade, meanwhile the bilirubin was reduced to 2 times the normal value. The GV/SLV ratios of 3 patients with low GV/SLV ratio had reached 44.4%, 47.2% and 56.2% respectively. In this study, the GV/SLV ratios of the 12 patients were between 73.2% and 40.8% with an average of 55.6%. Operation time was 11.5-20.5 h, with an average of 12.3 h. Anhepatic phase time was 193-375 min with median 253.5 min. The red blood cell suspension was 0-6 U during the operation. The average hospitalization was 10-42 d, with the average 22.7 d. Total hospital costs were 121 600-434 800 Yuan, with the median cost of 174 400 Yuan. One patient died of septic shock a week after surgery. Conclusion: (1)ALT may provide feasibility for the advanced HAE. (2)Accurate assessment of functional liver "volume and quality" appeared as the key points to the ALT. (3)Precise surgery and individualized treatment could improve and protect the functional liver "volume and quality" .

Chloroplast analysis of Zelkova schneideriana (Ulmaceae): genetic diversity, population structure, and conservation implications.

Zelkova schneideriana is endemic to China and belongs to the Ulmaceae. It is listed as a Near Threatened species in the China Biodiversity Red Data Book. We conducted a phylogeographical study of two chloroplast regions (psbA-trnH and trnG-trnM) in several Chinese Z. schneideriana populations, in order to examine the genetic diversity, population structure, and evolutionary history of the species. In all, 10 haplotypes were detected. The population from Sangzhi, Hunan, had the highest nucleotide diversity (π = 0.00653) and haplotype diversity (HD = 1.000), and should be considered the most suitable population to be protected under an in situ conservation strategy. Seed collections from as many individuals as possible in other populations would preserve the genetic diversity of Z. schneideriana.

An evaluation of zinc bearing palygorskite inclusion on the growth performance, mineral content, meat quality, and antioxidant status of broilers.

The current study was conducted to investigate the effect of zinc (Zn) bearing palygorskite (ZnPal) inclusion on the growth performance, mineral content, meat quality, and antioxidant status of broilers. A total of 240 one-day-old Arbor Acres broiler chicks were randomly allocated into 5 dietary treatments with 6 replicates of 8 chicks. Broilers in the 5 treatments were fed a basal diet supplemented with 0, 20, 40, 60, and 80 mg/kg Zn diet in the form of ZnPal for 42 d, respectively. Birds exhibited similar average daily gain (ADG), average daily feed intake (ADFI), and feed/gain ratio (F:G) among groups during the 42-day study (P>0.05). ZnPal supplementation linearly increased iron (Fe) (P=0.031) and magnesium (Mg) (P=0.002) content in the pectoralis major muscle. Similarly, the inclusion of ZnPal tended to increase Zn content in the thigh (P=0.072) and linearly increase Zn content in the pectoralis major muscle (P=0.055). The concentration of copper (Cu) in the thigh was linearly decreased by ZnPal inclusion (P=0.011). Meanwhile, a quadratic trend for reduced Cu content was observed in the pectoralis major muscle (P=0.074) and thigh (P=0.082), respectively. The supplementation of ZnPal linearly reduced cooking loss in the pectoralis major muscle (P=0.013), and linearly (P=0.029) and quadratically (P=0.034) decreased cooking loss in the thigh. Malondialdehyde (MDA) concentration in the thigh was linearly (P=0.020) and quadratically (P=0.017) reduced by ZnPal inclusion. Additionally, ZnPal supplementation tended to linearly enhance total antioxidant capacity (T-AOC) activity of the pectoralis major muscle (P=0.083). The results obtained in the current study indicated that ZnPal inclusion could alter muscular mineral accumulation, improve meat quality, and enhance the muscular antioxidant capacity of broilers, and Zn supplementation in the form of ZnPal at the dosage of 20 mg/kg would be sufficient in improving meat quality and muscular oxidative status.

[Living donor liver transplantation for the treatment complicated hepatic cystic echinococcosis: first case report from China].

OBJECTIVE: To report the first clinical experience of living donor liver transplantation(LDLT) in patient with complicated hepatic cystic echinococcosis in China. METHODS: First Affiliated Hospital of Xinjiang Medical University successfully implemented LDLT in the treatment of one patient with complicated hepatic cystic echinococcosis after four times liver surgery and secondary congestive cirrhosis on May 9, 2015. The clinical data of the patient has been retrospectively analyzed. RESULTS: LDLT procedure was successfully performed in a patient with four previous liver intervention due to his recurrence. The patient was lack of right lobe, while with significant hypertrophy in left lobe. The reoccurred lesion compressed the retrohepatic vena cava, left hepatic vein, portal vein, that led to the stricture of retrohepatic vena cava, left hepatic vein orifice. The phlebography displayed remarkable stricture in retrohepatic vena cava and rich collateral circulation by which the venous blood flow back to the right atrium via paravertebral vein. The orifice of left hepatic vein shaped as hair-like so that the stenting was impossibly. After the multi-disciplinary team discussion, the liver transplantation was proposed, while the autotransplantation was considered as first line option, and with the backup of living related donor. The laparotomy showed the hugely enlarged liver with nodular changes. The frozen section was tested twice and showed as spotty necrosis with pseudo-lobe formation. Although the enlarged left lobe about 2 000 g weight, the liver colored as grey with cotton-like texture, the plan for autotransplantation was abandoned regarding the high risk for post-operative liver failure. Therefore, LDLT was performed. The donor was her brother who donated right lobe with 685 g weight. The operation time was 1 005 min with anhepatic time 335 min. Total of 12 units red blood cell suspension was transfused. The coagulating function was back to normal in 3 days after operation. The patient was discharged at 30 days after operation, while the routine blood test and biochemical markers had back to normal levels. The patient had taken tacrolimus and methylprednisolone with normal life. The liver functions of transplanted liver were normal in 90 days after surgery. The patient was able to self-activity and regular outpatient follow-up. CONCLUSION: Hepatic CE is an infectious disease, and liver transplantation could be the last choice for complicated case, especially in combination with severe complications, like Budd-Chiari syndrome.

Isolation and characterization of polymorphic microsatellite loci from Zelkova schneideriana Hand.-Mazz.

Zelkova schneideriana is a highly valued hardwood species. An improved technique for isolating codominant compound microsatellite markers was used to develop simple sequence repeat markers for Z. schneideriana. A total of 12 microsatellite loci were identified. Overall, the number of alleles per locus ranged from 8-19, with an average of 11.75. Observed heterozygosity and expected heterozygosity values ranged from 0.109-0.709 and 0.832-0.929, respectively. Polymorphic information content is from 0.803-0.915, with an average of 0.854. These markers will be very important for future research related to the genetic diversity, population structure, patterns of gene flow, and mating system of this species.

[Anemia incidence and influencing factors among children aged 0-6 years in Shannan city, Xizang Autonomous Region].

Objective: To investigate the prevalence and influencing factors of anemia children aged 0-6 years in Shannan city, Xizang Autonomous Region. Methods: A cross-sectional study was performed to measure hemoglobin of 12 405 children aged 0-6 years in 12 counties (districts) of Shannan city, Xizang Autonomous Region and basic information including gender, age, ethnicity, address, local altitude, urban area, agricultural and pastoral areas, etc were collected. According to the altitude of the surveyed children's place of residence, anemia in children under the plain standard and the prevalence of anemia after adjusting for altitude were reported, and the influencing factors of anemia in these children were analyzed by Logistic regression methods. Results: Among the 12 405 children, 6 383 (51.46%) were males, 6 022 (48.54%) were females, 462 (3.72%) were newborns, 68 (0.55%) were 1 month old children, 153 (1.23%) were 4-<6 months old children, and 11 722 (94.49%) were 6 months to 6 years old children. The prevalence of anemia in children aged 0-6 years in Shannan city before altitude correction was 15.50%(1 923/12 405), and it was 57.84% (7 175/12 405) after altitude correction. The hemoglobin level in the children was (131±20) g/L. Univariate analysis showed that the anemia rate of males was higher than that of females (59.17% (3 777/6 383) vs.56.43% (3 398/6 022), P<0.05), the anemia rate of Tibetan children was higher than that of children of other ethnic groups (58.21% (6 992/12 011) vs.46.45% (183/394), P<0.05), the anemia rate of children in agricultural and pastoral areas was higher than that of urban children (58.75% (6 212/10 574) vs.(52.59% 963/1 831), P<0.05), and the anemia rate of children in higher-altitude areas was higher than that of children in high-altitude areas (67.98% (2 773/4 079) vs.52.87% (4 402/8 326), P<0.05). Logistic regression analysis revealed that girls (OR=0.89, 95%CI 0.83-0.96), children of other ethnic groups (OR=0.72, 95%CI 0.58-0.89), children aged 4-<6 months (OR=0.40, 95%CI 0.24-0.67) and children aged 6 months to 6 years (OR=0.59, 95%CI 0.43-0.82) were protective factors, and children who lived in agricultural and pastoral areas (OR=1.15, 95%CI 1.04-1.28) and children living at altitudes higher than 4 000 m areas (OR=1.88, 95%CI 1.73-2.04) were independent risk factors for the occurrence of anemia. Conclusions: The prevalence of anemia in children aged 0-6 years in Shannan city was still relatively high. It might be affected by factors such as sex, ethnicity, age of the children, place of residence and altitude where the children lived.

[Incidence of common gene mutations in early-onset colorectal cancer and the association with cancer survival: a meta-analysis].

Objective: The incidence of early-onset colorectal cancer (EOCRC) is increasing globally; however, the molecular characteristics and prognosis of sporadic EOCRC are unclear. In this systematic review and meta-analysis, we aimed to investigate the incidence of gene mutations and their association with cancer survival in sporadic EOCRC, focusing on six common gene mutations (TP53, BRAF, KRAS, NRAS, PTEN, and APC). Methods: Ovid Embase and Ovid Medline electronic databases were searched for studies involving patients with sporadic EOCRC (i.e., diagnosed with colorectal cancer before the age of 50 years and with no evidence of hereditary syndromes predisposing to colorectal cancer). The included articles were evaluated using quality assessment tools. Meta-analysis was performed using random-effects and fixed-effects models. Cochran's Q statistic and the I2 index were used to assess heterogeneity. The incidence of the six common gene mutations listed above in sporadic EOCRC and their association with cancer survival were evaluated. Results: (1) Incidence of specific gene mutations in sporadic EOCRC. A total of 34 articles were included in this meta-analysis. The incidence of APC gene mutation was 36% (from 13 articles, 95%CI: 19%-55%, P=0.043); of KRAS gene mutation 30% (from 26 articles, 95%CI: 24%-35%, P=0.190); of BRAF gene mutation 7% (from 18 articles, 95%CI: 5%-11%, P=0.422); of NRAS gene mutation 4% (from five articles, 95%CI: 3%-5%, P=0.586); of PTEN gene mutation 6% (from six articles, 95%CI: 4%-10%, P=0.968); and of TP53 gene mutation 59% (from 13 articles, 95%CI: 49%-68%, P=0.164). (2) Association between gene mutations and survival in sporadic EOCRC. A total of six articles were included in this meta-analysis. Compared with wild-type BRAF, mutant BRAF was significantly associated with increased overall mortality risk in patients with EOCRC (pooled HR=2.85, 95%CI: 1.45-5.60, P=0.002). Subgroup analysis showed that the incidence of BRAF gene mutation was higher in Eastern than in Western countries, whereas the incidence of TP53, KRAS, NRAS, and APC gene mutations was lower. There was no significant difference in the incidence of PTEN gene mutation between different regions. Conclusion: Compared with colorectal cancer occurring in the general population, the incidence of APC and KRAS mutations is lower in EOCRC, whereas the incidence of TP53 mutation remains consistent. BRAF mutation is associated with increased overall mortality risk in patients with EOCRC.

Superposed pulse amplitude modulation for visible light communication.

We propose and experimentally demonstrate a novel modulation scheme called superposed pulse amplitude modulation (SPAM) which is low-cost, insensitive to non-linearity of light emitting diode (LED). Multiple optical pulses transmit parallelly from different spatial position in the LED array and overlap linearly in free space to realize SPAM. With LED arrangement, the experimental results show that using the modulation we proposed the data rate of 120 Mbit/s with BER 1 × 10(-3) can be achieved with an optical blue filter and RC post-equalization.

Mitigation of CO poisoning on functionalized Pt-TiN surfaces.

It has been previously reported that the system of single Pt atoms embedded in N-vacancy (V(N)) sites on the TiN(100) surface (Pt-TiN) could be a promising catalyst for proton exchange membrane fuel cells (PEM FCs). The adsorption of molecules on Pt-TiN is an important step, when it is incorporated as the anode or cathode of PEM FCs. Utilizing first principles calculations based on density functional theory, systematic investigations are performed on the adsorption of several atomic and molecular species on the Pt-TiN system, as well as the co-adsorption of them. The favorable binding sites and adsorption energies of several molecular species, namely carbon dioxide (CO2), carbon monoxide (CO), oxygen (O2), hydrogen (H2), hydroxyl (OH), an oxygen atom (O), and a hydrogen atom (H), are explored. For each, the adsorption energy and preferred binding site are identified and the vibrational frequencies calculated. It is found that CO2, CO and H prefer the Pt top site while OH and O favorably adsorb on the Ti top site. When CO and OH are co-adsorbed on the Pt-TiN(100) surface, OH weakens the adsorption of CO. The weakening effect is enhanced by increasing the coverage of OH. A similar behavior occurs for H and OH co-adsorption on the Pt-TiN(100) surface. Because co-adsorption with OH and H species weakens the adsorption of CO on Pt-TiN, it is expected that the acid and base conditions in PEM FCs could mitigate CO poisoning on functionalized Pt-TiN surfaces.

Rhizoctonia solani Identified as the Disease Causing Agent of Peanut Leaf Rot in China.

Peanut (Arachis hypogaea L.) is one of the most economically important oil crops in the world. Since the 1990s, the peanut industry has developed rapidly in China. However, because of the use of high-yield varieties and increased plant density, a peanut leaf rot disease occurred in Laixi Experimental Fields in Shandong Province, China in 2007. Leaves had nearly circular, brown lesions that enlarged quickly developing yellow-brown halos at the edges of the lesions. High relative humidity under field conditions led to complete necrosis of the leaves with cotton wool-like mycelia observed followed by the development of sclerotia on the leaf surface. Symptomatic plants were observed between 2007 and 2010, and symptomatic leaf tissue was collected from the Laixi Experimental Fields. An isolate (designated YF-1) from symptomatic peanut leaves was isolated and purified on potato dextrose agar (PDA) and water agar (WA) medium. On PDA, the colony appeared initially as colorless and grew to the diameter of a 9-cm petri dish within 3 days. As the mycelium aged, the colony color gradually became light brown, and sclerotia developed on the surface of the colony. YF-1 was identified as Rhizoctonia solani Kühn based on the number of nuclei per cell ranging from 4 to 13 (average 6.1), hyphal diameter being 7.5 to 12.9 µm (average 8.3 µm), branching at right angles, a septum was present near each hyphal branch with a slight constriction, and no clamp connection structures or conidia were ever observed (4). To further confirm the identity of isolate YF-1, genomic DNA was extracted using the DNeasy Plant Mini DNA Extraction Kit (Shanghai Leifeng Biotechnol. Co., Ltd.), and the complete internal transcribed spacer (ITS) region of ribosomal DNA was amplified and sequenced with a pair of primers ITS1/ITS4 (2). A GenBank BLAST search produced an exact match for the sequences of R. solani (AY154301), with 100% sequence similarity. To estimate the mode of anastomosis, YF-1 was paired on WA medium with each reference strain belonging to anastomosis groups (AGs) 1 through 8 (provided by Shandong Agriculture University) (1,3). The results indicated that YF-1 belonged to group AG-1, subgroup AG-1-IA of R. solani. Pathogenicity tests were conducted by inoculating 10 peanut leaves using a colonized paper disc method (filter paper 1 cm in diameter suspended in the mycelia suspension). Ten control leaves received paper discs without mycelium. Inoculated and non-inoculated plants were kept in humid chambers for 24 h at 25°C. Three days after inoculation, the leaves developed typical brown lesions that were similar to those of naturally diseased plants. Koch's postulates were fulfilled by reisolation of R. solani from symptomatic leaves. No symptoms were observed on control leaves. To our knowledge, this is the first report of peanut leaf rot caused by R. solani. Occurrence of the disease in China is a new threat to the health of peanut. References: (1) Y. X. Chen et al. Acta Phytopathol. Sin. 3:139, 1985. (2) T. Misawa and S. Kuninaga. J. Gen. Plant Pathol. 76:310, 2010. (3) A. Ogoshi. Ann. Phytopathol. Soc. Jpn. 38:117, 1972. (4) J. R. Jr. Pameter and H. S. Whitmey. UC Press. 135, 1970.

Enhanced removal of chemical oxygen demand, nitrogen and phosphorus using the ameliorative anoxic/anaerobic/oxic process and micro-electrolysis.

Synthetic wastewater was treated using a novel system integrating the reversed anoxic/anaerobic/oxic (RAAO) process, a micro-electrolysis (ME) bed and complex biological media. The system showed superior chemical oxygen demand (COD), total nitrogen (TN) and total phosphorus (TP) removal rates. Performance of the system was optimised by considering the influences of three major controlling factors, namely, hydraulic retention time (HRT), organic loading rate (OLR) and mixed liquor recirculation (MLR). TP removal efficiencies were 69, 87, 87 and 83% under the HRTs of 4, 8, 12 and 16 h. In contrast, HRT had negligible effects on the COD and TN removal efficiencies. COD, TN and TP removal efficiencies from synthetic wastewater were 95, 63 and 87%, respectively, at an OLR of 1.9 g/(L·d). The concentrations of COD, TN and TP in the effluent were less than 50, 15 and 1 mg/L, respectively, at the controlled MLR range of 75-100%. In this system, organics, TN and TP were primarily removed from anoxic tank regardless of the operational conditions.