[Value of double contrast-enhanced ultrasound QontraXt three-dimensional pseudocolor quantitative analysis to therapeutic effect evaluation of preoperative neoadjuvant chemotherapy in advanced gastric cancer patients].

Objective: To investigate the value of tumor perfusion parameter measured by using double contrast-enhanced ultrasound (DCEUS) QontraXt three-dimensional pseudocolor quantitative analysis to the therapeutic effect evaluation of preoperative neoadjuvant chemotherapy (NAC) in advanced gastric cancer (AGC) patients. Methods: Eighty-nine AGC patients underwent 3 cycles of preoperative NAC (XELOX) followed by complete resection of lesion. The DCEUS QontraXt three-dimensional pseudocolor was performed one or two weeks before the NAC and operation were applied, respectively. The peak enhancement (PE), time to peak (TP), sharpness of the bolus (ß) and area under the enhancement curve (AUC) of primary gastric tumor were measured by QontraXt three-dimensional pseudocolor quantitative analysis. These DCEUS parameters between respond and non-respond groups before and after NAC therapy were compared. The prediction accuracy of DCEUS to the therapeutic effect evaluation of preoperative NAC was determined by the receive operating characteristic (ROC) curves. Results: Among 89 AGC patients, 52 patients responded to NAC therapy, while 37 patients resisted to NAC therapy. Twelve cases in respond group and 26 cases in non-respond group were mucinous carcinoma. Forty cases in respond group and 11 cases in non-respond group were non-mucinous carcinoma (P<0.05). In responder group, the PE and TP before NAC were (53.7±9.3)% and (14 521±2 667) ms, and (32.2±5.5)% and (17 235±1 898) ms after NAC. The ratio of changes of PE (ΔPE) and TP (ΔTP) were 0.43±0.17 and 0.36±0.14, respectively. In non-respond group, the PE and TP before NAC were (54.4±7.2)% and (13 869±3 247) ms, and (45.3±6.1)% and (15 127±1 423) ms after NAC therapy. The ratio of ΔPE and ΔTP were 0.24±0.20 and 0.22±0.12. The PE and TP after NAC, the ratio of ΔPE and ΔTP were significant different among these two groups (all of P<0.05). The ROC curves showed that the ratio of ΔPE in assessing the respond of gastric cancer patients to NAC was superior compared to other parameters (AUC=0.784, P=0.004). The optimal cut-off value of the ratio of ΔPE was 24% and its sensitivity and specificity to the therapeutic effect evaluation of NAC in gastric cancer were 82.7% and 64.9%. Conclusion: DCEUS QontraXt three-dimensional pseudocolor quantitative analysis might be a novel, noninvasive and reliable method to evaluate the therapeutic effect of preoperative NAC in AGC patients.

[Comparison of aortic annular diameter defined by different measurement mordalities before transcatheter aortic valve implantation].

Objective: To compare aortic annular diameter measured by transthoracic echocardiography (TTE), transesophageal echocardiography (TEE), and multislice computed tomography (MSCT) in patients with severe aortic stenosis, and to evaluate the impact on selection of prosthetic valve type in transcatheter aortic valve implantation (TAVI). Methods: Clinical data of 138 patients with severe aortic stenosis referred for TAVI between January 2014 and June 2016 in our hospital were retrospectively analyzed.The difference of aortic annular diameter measured by TTE, TEE, and MSCT were compared.TTE was performed after TAVI to evaluate the accuracy of measurement before TAVI. Results: (1) Aortic annular diameter was (23.37±2.22) mm by TTE and (23.52±1.70) mm by TEE (P=0.12). Pearson correlation analysis showed that aortic annular diameter measured by TTE was correlated to that measured by TEE (r=0.87, P<0.05). (2)The long-axis diameter and the short-axis diameter measured by MSCT multiplanar reconstruction were significantly different ((27.86±2.87) mm vs. (21.91±2.53) mm, P<0.05). There was no significant difference between the mean of the long- and short-axis diameters and the diameter derived from cross-sectional area by MSCT ((24.92±2.38) mm vs. (24.84±2.25) mm, P=0.08). However, the diameter derived from the circumference ((25.35±2.34) mm) was significantly larger than the mean of the long- and short-axis diameters and the diameter derived from cross-sectional area by MSCT, and the difference were (0.43±0.62) mm and (0.51±0.62) mm respectively(both P<0.05). (3) Measurements of the aortic annulus diameter by MSCT including the mean of the long- and short-axis diameters, diameter derived from cross-sectional area, and diameter derived from the circumference were larger than the measurement by TTE and TTE (all P<0.05). (4) Implantation was successful in all patients.Moderate to severe paravalvular leakage was detected in 3 patients at 7 days post TAVI, and 1 patient developed severe prosthetic valve restenosis at 6 months post TAVI and received valve-in-valve implantation. Conclusions: In severe aortic stenosis patients referred for TAVI, the aortic annular diameter derived from TTE and TEE measurements are smaller than that from MSCT.In the absence of a gold standard, selection of prosthetic valve type in TAVI procedure should rely on comprehensive considerations, which is of importance to get good clinical results for severe aortic stenosis patients underwent TAVI.

Performance of risk estimation for hepatocellular carcinoma in chronic hepatitis B (REACH-B) score in classifying treatment eligibility under 2012 Asian Pacific Association for the Study of the Liver (APASL) guideline for chronic hepatitis B patients.

BACKGROUND: REACH-B [Risk Estimation for Hepatocellular Carcinoma (HCC) in Chronic Hepatitis B] scoring system was developed to predict the risk of HCC in noncirrhotic chronic hepatitis B (CHB) patients. AIM: To evaluate the discriminatory performance of REACH-B scoring system in classifying anti-viral treatment eligibility of CHB patients according to the 2012 Asian Pacific Association for the Study of the Liver (APASL) treatment guideline. METHODS: A total of 904 noncirrhotic CHB were enrolled. Patients' age, gender, liver biochemistry, HBeAg status and HBV DNA levels were recorded. RESULTS: The minimum REACH-B risk score for patients to be eligible for anti-viral treatment was 7 for HBeAg-positive and 6 for HBeAg-negative patients. Among them, increasing REACH-B score was not significantly associated with eligibility for treatment [adjusted odds ratio (OR): 1.210, 95% confidence interval (CI): 0.979-1.494, P = 0.078] in HBeAg-positive patients, as shown by logistic regression analysis after adjusting for gender. In HBeAg-negative patients, REACH-B score significantly predicted the treatment eligibility (adjusted OR: 1.783, 95% CI: 1.607-1.979, P < 0.001). Discriminatory ability of REACH-B score to classify eligibility was poor for HBeAg-positive patients ≥40 years [area under receiver operating characteristic (AUC): 0.664, 95% CI: 0.533-0.795], but good/excellent for HBeAg-positive patients <40 years (AUC: 0.903; 95% CI: 0.841-0.964), HBeAg-negative patients ≥45 years (AUC: 0.883; 95% CI: 0.848-0.917) and HBeAg-negative patients <45 years (AUC: 0.907; 95% CI: 0.874-0.940). CONCLUSION: The discriminatory performance of the REACH-B scoring system in classifying anti-viral treatment eligibility based on the 2012 APASL guideline was good/excellent, except for ≥40 years old HBeAg-positive patients.

Isolation of biosurfactant producers, optimization and properties of biosurfactant produced by Acinetobacter sp. from petroleum-contaminated soil.

AIMS: To screen and identify biosurfactant producers from petroleum-contaminated soil; to use response surface methodology (RSM) for medium optimization to enhance biosurfactant production; and to study the properties of the newly obtained biosurfactant towards pH, temperature and salinity. METHODS AND RESULTS: We successfully isolated three biosurfactant producers from petroleum-contaminated soil and identified them through 16S rRNA sequence analysis, which exhibit the highest similarities to Acinetobacter beijerinckii (100%), Kocuria marina (99%) and Kineococcus marinus (99%), respectively. A quadratic response model was constructed through RSM designs, leading to a 57·5% increase of the growth-associated biosurfactant production by Acinetobacter sp. YC-X 2 with an optimized medium: beef extract 3·12 g l(-1) ; peptone 20·87 g l(-1) ; NaCl 1·04 g l(-1); and n-hexadecane 1·86 g l(-1). Biosurfactant produced by Acinetobacter sp. YC-X 2 retained its properties during exposure to a wide range of pH values (5-11), high temperatures (up to 121°C) and high salinities [up to 18% (w/v) Na(+) and Ca(2+) ], which was more sensitive to Ca(2+) than Na(+). CONCLUSIONS: Two novel biosurfactant producers were isolated from petroleum-contaminated soil. Biosurfactant from Acinetobacter sp. YC-X 2 has good properties to a wide range of pH, high temperature and high salinity, and its production was optimized successfully through RSM. SIGNIFICANCE AND IMPACT OF THE STUDY: The fact, an increasing demand of high-quality surfactants and the lack of cost-competitive bioprocesses of biosurfactants for commercial utilization, motivates researchers to develop cost-effective strategies for biosurfactant production through isolating new biosurfactant producers with special surface-active properties and optimizing their cultural conditions. Two novel biosurfactant producers in this study will widen our knowledge about this kind of micro-organism. This work is the first application of RSM designs for cultural optimization of biosurfactant produced by Acinetobacter genus and the first report that biosurfactant may be more sensitive to Ca(2+) than Na(+) .

Enhancement of CPT-11 antitumor activity by adenovirus-mediated expression of ß-glucuronidase in tumors.

CPT-11 is a clinically important prodrug that requires conversion into the active metabolite SN-38, a potent topoisomerase I poison, for antitumor activity. However, SN-38 is rapidly metabolized to the inactive SN-38 glucuronide (SN-38G) in the liver, which reduces the amount of SN-38 available for killing cancer cells. Here, we investigated if local expression of ß-glucuronidase (ßG) on cancer cells to catalytically convert SN38G to SN38 could enhance the antitumor activity of CPT-11. ßG was tethered on the plasma membrane of three different human cancer cell lines: human colon carcinoma (LS174T), lung adenocarcinoma (CL1-5) and bladder carcinoma (EJ). Surface ß-glucuronidase-expressing cells were 20 to 80-fold more sensitive to SN-38G than the parental cells. Intravenous CPT-11 produced significantly greater suppression of CL1-5 and LS174 T tumors that expressed ßG as compared with unmodified tumors. Furthermore, an adenoviral vector expressing membrane-tethered ßG (Ad.ßG) increased the sensitivity of cancer cells to SN-38G even at multiplicity of infections as low as 0.16, indicating bystander killing of non-transduced cancer cells. Importantly, intratumoral injection of Ad.ßG significantly enhanced the in vivo antitumor activity of CPT-11 as compared with treatment with CPT-11 or Ad vectors alone. This study shows that Ad.ßG has potential to boost the therapeutic index of CPT-11.

Reappraisal of the importance of mutations in the NS5A-PKR-binding domain of hepatitis C-1b virus in the era of optimally individualized therapy.

Past studies have reported that mutations in the protein kinase R-binding domain (PKRBD) sequences of hepatitis C virus (HCV) NS5A proteins are correlated with response to fixed-duration interferon (IFN)-based therapy in patients infected with HCV-1b. In this study, we investigated whether the substitutions in PKRBD, including the IFN sensitivity-determining region (ISDR) and 26 additional downstream amino acids from ISDR, will have effects upon patients infected with chronic HCV-1b in the era of individualized therapy with peginterferon and ribavirin. Thirty-seven patients were treated with optimally tailored therapy guided by baseline viral load combined with rapid and early virological responses while 23 patients were treated without guidance and/or assigned suboptimal treatment duration. The amino acid sequences of the PKRBD were determined by PCR and sequencing. The overall sustained virological response (SVR) rate of patients who received optimally individualized therapy was 78.4%, which was better than the SVR rate of patients who received suboptimal therapy (47.8%, P = 0.015). Multivariate analysis showed that optimally individualized therapy (P = 0.019) and 80/80/80 adherence (P = 0.006) were independent favourable predictors of SVR in the entire cohort. Further sub-analysis of the predictive factors of SVR in patients treated with optimally individualized therapy showed that mutations in the 26-amino acid downstream from the ISDR (P = 0.024) were the only independent predictor of SVR. We concluded that mutations in 26-amino acid downstream portion from the ISDR remained a prognosticator of SVR in the era of optimally tailored therapy.

Development of recombinant adeno-associated virus and adenovirus cocktail system for efficient hTERTC27 polypeptide-mediated cancer gene therapy.

The low in vivo transduction efficiency of recombinant adeno-associated virus (rAAV) and the undesirably strong immunogenicity of adenovirus (rAdv) have limited their clinical utilization in cancer gene therapy. We have previously demonstrated that intratumoral injection of rAAV expressing a C-terminal polypeptide of human telomerase reverse transcriptase (rAAV-hTERTC27) effectively inhibits the growth of glioblastoma xenografts in nude mice. To further improve its efficacy, we combined rAAV-hTERTC27 with rAdv and investigated the efficiency of the cocktail vectors in vivo. At a nontherapeutic dose (1 x 10(8) plaque-forming units (PFUs)), rAdv-null and rAdv-hTERTC27 were equipotent in enhancing the therapeutic efficacy of rAAV-hTERTC27 (1.5 x 10(11) v.g.), and complete tumor regression was achieved in 25% of the treated animals. Importantly, the combination of rAAV-hTERTC27 and a therapeutic dose (2.5 x 10(9) PFU) of rAdv-hTERTC27 significantly augmented the therapeutic effects and led to a 38% complete tumor regression rate. In vivo optical imaging also showed that rAAV-luc/rAdv-luc cocktail vectors could synergistically enhance the early transient and latent sustained expression of luciferase, as compared to rAdv-luc and rAAV-luc alone. These findings suggest that the combination of rAAV-hTERTC27 and a therapeutic dose of rAdv-hTERTC27 is potentially a promising treatment for glioblastoma, and the rAAV/rAdv cocktail vector system warrants further development for cancer gene therapy.

A novel glioblastoma cancer gene therapy using AAV-mediated long-term expression of human TERT C-terminal polypeptide.

Glioblastoma multiforme is the most aggressive form of human brain tumor, which has no effective cure. Previously, we have demonstrated that overexpression of the C-terminal fragment of the human telomerase reverse transcriptase (hTERTC27) inhibits the growth and tumorigenicity of human cervical cancer HeLa cells. In this study, the therapeutic effect and molecular mechanisms of hTERTC27-mediated cancer gene therapy were further explored in vivo in established human glioblastoma xenografts in nude mice. We showed that intratumoral injection of adeno-associated virus carrying hTERTC27 (rAAV-hTERTC27) is highly effective in reducing the growth of the subcutaneously transplanted glioblastoma tumors. Histological analyses showed that rAAV-hTERTC27 treatment leads to profound necrosis, apoptosis, infiltration of polymorphonuclear neutrophils and reduced microvessel density in the tumor samples. To study the molecular mechanism of rAAV-hTERTC27-mediated antitumor effects, we analyzed the global gene expression profiles of the rAAV-hTERTC27-treated tumor tissues and cell line as compared with that of the control rAAV-green fluorescent protein-treated samples by DNA microarray. Our results suggest that hTERTC27 exerts its effect through complex mechanisms, which involve genes regulating apoptosis, cell adhesion, cell cycle, immune responses, metabolism, signal transduction, transport, transcription and telomere maintenance.

Suspended aggregates as an immobilization mode for high-density perfusion culture of HEK 293 cells in a stirred tank bioreactor.

Cells of the human embryonic kidney cell line (HEK 293) grown in repeated suspension and perfusion systems were characterized and described. Cell aggregates that formed immediately after the HEK 293 cells were inoculated in stirred vessels in serum-containing Dulbecco's modified Eagle's medium (D-MEM)/F-12 medium. The mean diameter of the cell aggregates reflecting the aggregate size increased with culture time, shifting from 63 to 239 mum after 1 and 8 days of culture in spinner flasks, respectively. No significant differences in cell performance were observed between HEK 293 cell populations grown as suspended aggregates and those grown as anchored monolayers. Replacing the D-MEM/F-12 with CD 293 medium caused the compact spherical cell aggregates to dissociate into single cells and small irregular aggregates without any apparent effect on cell performance. Moreover, the spherical cell aggregates could reform from individual cells and small aggregates when exposed to the serum-containing D-MEM/F-12 dominant medium. Perfusion culture of HEK 293 cells grown as suspended aggregates in a 7.5-l stirred tank bioreactor for 17 days resulted in a maximum viable cell density of 1.2 x 10(7) cells ml(-1). These results demonstrate the feasibility and proof-of-concept for using aggregates as an immobilization system in large-scale stirred bioreactors because a small-scale culture can be used as easily as the inoculum for larger bioreactors.

Effects of fertilization promoting peptide, adenosine, and pentoxifylline on thawed human sperm.

Fertilization promoting peptide (FPP) and adenosine were demonstrated to be potential modulators of sperm capacitation in mammals. Both FPP and adenosine, by modulating the adenylate cyclase (AC)/cAMP signaling pathway, elicit similar biphasic responses in mammalian sperm (i.e., stimulating capacitation and inhibiting spontaneous acrosome loss). Pentoxifylline, an artificial sperm stimulant, is clinically used to enhance motility of sperm from infertile men. By inhibiting phosphodiesterase, pentoxifylline increases the intracellular cAMP level of sperm, and thus contributes to capacitation, hyperactivation, and acrosome reaction in animal studies. The effects of FPP, adenosine, and pentoxifylline on thawed human sperm are stressed. Chlortetracycline (CTC) fluorescence assessment revealed that none of the 3 reagents improved fertilization ability of post-thawed sperm. Motility studies with computer-aided sperm analyzer (CASA) showed significantly smaller STR (straight-line velocity) and LIN (linearity) in the FPP-treated group at 4 h of incubation p

Structural influence of hanatoxin binding on the carboxyl terminus of S3 segment in voltage-gated K(+)-channel Kv2.1.

The voltage-sensing domains of voltage-gated potassium channels Kv2.1 (drk1) contain four transmembrane segments in each subunit, termed S1 to S4. While S4 is known as the voltage sensor, the carboxyl terminus of S3 (S3C) bears a gradually broader interest concerning the site for gating modifier toxins like hanatoxin and thus the secondary structure arrangement as well as its surrounding environment. To further examine the putative three-dimensional (3-D) structure of S3C and to illustrate the residues required for hanatoxin binding (which may, in turn, show the influence on the S4 in terms of changes in channel gating), molecular simulations and dockings were performed. These were based on the solution structure of hanatoxin and the structural information from lysine-scanning results for S3C fragment. Our data suggest that several basic and acidic residues of hanatoxin are electrostatically and stereochemically mapped onto their partner residues on S3C helix, whereas some aromatic or hydrophobic residues located on the same helical fragment interact with the hydrophobic patch of the toxin upon binding. Therefore, a slight distortion of the S3C helix, in a direction toward the N-terminus of S4, may exist. Such conformational change of S3C upon toxin binding is presented as a possible explanation for the observed shift in hanatoxin binding-induced gating.

Effectiveness of postoperative adjuvant therapy in improving reproductive outcome of endometriosis-associated infertility.

BACKGROUND AND PURPOSE: Treatment of endometriosis-associated infertility has not yet become standardized. Various protocols including surgical treatment, medical therapy, and a combination of both have been suggested but their use remains controversial. The objective of the present study was to determine whether postoperative adjuvant therapy for endometriosis is effective in improving reproductive outcome. METHODS: Medical records of infertile patients with newly diagnosed endometriosis treated in a university teaching hospital during a 50-month period were reviewed. After exclusion of patients with other major infertility factors, a total of 209 patients were included in the retrospective analysis. These patients were divided into those receiving (n = 78) or not receiving (n = 131) peri- or postoperative adjuvant medical therapy. The adjuvant therapies included danazol (n = 62), gonadotropin releasing hormone analogues (n = 11), progestins (n = 3), oral contraceptives (n = 1), and mixed treatment (n = 1). RESULTS: The pregnancy rate was lower in those receiving adjuvant therapy, although this result was not significant (32.1% vs 45.8%; p = 0.05). When patients using postoperative danazol therapy were considered alone, the pregnancy rate in patients receiving adjuvant therapy was significantly lower than that in patients not receiving it (p = 0.047). When the stage of endometriosis was considered, the pregnancy rate in patients receiving adjuvant therapy was again lower than in those not receiving it in patients with minimal or mild endometriosis (42.9% vs 60%; p = 0.043). However, in patients with moderate or severe endometriosis, the pregnancy rate was not different in the two groups (31% vs 36%; p = 0.56). Postoperative assisted reproductive techniques (ART) including controlled ovarian hyperstimulation/intrauterine insemination (COH/IUI) and in vitro fertilization (IVF) were effective in improving the pregnancy rates for all patients (53.9% with ART vs 33.1% without; p = 0.003) and for patients with advanced endometriosis (47.7% with ART vs 27.2% without; p = 0.016). CONCLUSIONS: Our results suggest that postoperative adjuvant therapy is ineffective in improving reproductive outcome in patients with either early (minimal or mild) or advanced (moderate and severe) endometriosis. This finding suggests that if fertility is the goal of treatment, adjuvant therapy may be unnecessary after surgery. In contrast, our data suggest that empirical ART, including COH/IUI or IVF, may be a better alternative to improve the pregnancy outcome after surgery.

[Bovine alpha-sl-casein gene sequences direct expression of a variant of human tissue plasminogen activator in the milk of transgenic mice].

The fusion gene containing the promoter of casein gene, LAtPA minigene and 3' flanking sequences of casein gene was introduced into the fertilized eggs of mice by microinjection. Five positive transgenic mice were obtained. The concentration of LAtPA in the milk of one female transgenic mouse is 0.18 microgram/ml. This result showed that the LAtPA minigene could correctly express the bioactive LAtPA in the milk of the transgenic mouse under the control of the regulatory elements of the casein gene.

[New advances and future perspectives in mammal cloning].

To the great extent, the study and application of transgenic animal are restricted by the inherent limitation of pronuclear microinjection. Recently, the rapid progresses in gene targeting and cloning of somatic cells have shown that the combination of these two technologies will become a virtual way to producing large transgenic animals.

[The expression of tPA directed by the bovine BLG regulatory elements in the mammary gland of transgenic mice].

In order to get the regulatory elements which are essential for generating mammary gland bioreactors, the whole 8.4 kb bovine BLG gene was obtained by PCR amplification. The 1.6 kb chicken lysozyme matrix attachment region (MAR) was used to overcome position effects. The bovine BLG-tPA expression vector was constructed and the BLG-tPA fusion gene was introduced into fertilized eggs of mice by microinjection to generate transgenic mouse. 170 offsprings were obtained, of which 9 were proved to be transgenic mice based on PCR and Southern-blot analysis. The tPA expression level amounted to 12 micrograms/mL in the milk of mice. The bovine BLG-tPA fusion gene integrated in the founders was inheritable.

[High cell density culture of phosphotransacetylase mutants of Escherichia coli BL21(DE3)].

Cell culture, organic acid production and foreign protein (TNF) expression of E. coli BL21(DE3) and its pta mutant were investigated. Under shaking conditions, TNF expression in pta mutant increased by 23%. During the fed-batch culture without limitation of specific growth rates, the mutant reached a cell density as high as 32.5 g(DCW)/L and total TNF expression at 2.8 g/L, while the parental strain only obtained 19.5 g(DCW)/L and 0.84 g/L. The results indicate that utility of pta mutant as a host is advantageous in foreign protein expression and high cell density culture. Meanwhile, the analysis data of organic acids accumulated during fed-batch culture showed that as the decrease of acetate production(42% of the parental strain), the accumulation of other organic acids(mainly pyruvate, lactate and succinate) obviously increased. As a result, the amount of total organic acids increased by 123% over its parent. The lactate production may be the main obstacle in further growth of the cells.

[Co-integration of BLG-LAtPA and WAP improved the expression of LAtPA in transgenic mouse milk].

In order to improve the expression of longer acting tissue plasminogen activator in the mammary epithelium of transgenic mice, the fragment of BLG-LAtPA hydrid gene was microinjected into mouse embryos with mice whey acid protein gene. Three mouse were tested as being Co-integration of BLG-LAtPA and WAP transgene by PCR and Southern blot. Milk obtained from lactating females contains biologically active tPA, and the concentration of tPA was calculated to be about 10 micrograms/mL.

The feasible sequential control strategy of treating high strength organic nitrogen wastewater with sequencing batch biofilm reactor.

The bio-kinetics and feasible sequential control strategy of treating high strength organic carbon and nitrogen wastewater were investigated by conducting the ABS manufacturing wastewater in a series of Sequencing Batch Biofilm Reactors (SBBRs). The on-line ORP, pH, and DO monitoring parameters were applied to identify the feature-points when ammonification, nitrification, and denitrification ends. The carbonaceous matter removal kinetics in the anaerobic and aerobic reaction stages can be expressed by the Michaelis-Menten equation. High efficiency of organic carbon removal and organic nitrogen ammonification in the anaerobic stage can eliminate the substrate competition and activation inhibition to nitrifying organisms in the following aerobic stage. In the sequencing nitrogen removal processes, the producing time and system ORP values of these feature-points have good function relationships with the influent COD loading rates of SBBR, which can be integrated into a set-point (set-time and set-ORP) sequential control strategy of nitrogen removal. The automatic control operation results revealed ORP was one of the major control parameters of the sequencing nitrogen removal process in SBBR system and high overall removal efficiency were obtained.