Identification and characterization of a novel hydroxylamine oxidase, DnfA, that catalyzes the oxidation of hydroxylamine to N2.

Nitrogen (N2) gas in the atmosphere is partially replenished by microbial denitrification of ammonia. Recent study has shown that Alcaligenes ammonioxydans oxidizes ammonia to dinitrogen via a process featuring the intermediate hydroxylamine, termed "Dirammox" (direct ammonia oxidation). However, the unique biochemistry of this process remains unknown. Here, we report an enzyme involved in Dirammox that catalyzes the conversion of hydroxylamine to N2. We tested previously annotated proteins involved in redox reactions, DnfA, DnfB, and DnfC, to determine their ability to catalyze the oxidation of ammonia or hydroxylamine. Our results showed that none of these proteins bound to ammonia or catalyzed its oxidation; however, we did find DnfA bound to hydroxylamine. Further experiments demonstrated that, in the presence of NADH and FAD, DnfA catalyzed the conversion of 15N-labeled hydroxylamine to 15N2. This conversion did not happen under oxygen (O2)-free conditions. Thus, we concluded that DnfA encodes a hydroxylamine oxidase. We demonstrate that DnfA is not homologous to any known hydroxylamine oxidoreductases and contains a diiron center, which was shown to be involved in catalysis via electron paramagnetic resonance experiments. Furthermore, enzyme kinetics of DnfA were assayed, revealing a Km of 92.9 ± 3.0 µM for hydroxylamine and a kcat of 0.028 ± 0.001 s-1. Finally, we show that DnfA was localized in the cytoplasm and periplasm as well as in tubular membrane invaginations in HO-1 cells. To the best of our knowledge, we conclude that DnfA is the first enzyme discovered that catalyzes oxidation of hydroxylamine to N2.

Novel Alcaligenes ammonioxydans sp. nov. from wastewater treatment sludge oxidizes ammonia to N2 with a previously unknown pathway.

Heterotrophic nitrifiers are able to oxidize and remove ammonia from nitrogen-rich wastewaters but the genetic elements of heterotrophic ammonia oxidation are poorly understood. Here, we isolated and identified a novel heterotrophic nitrifier, Alcaligenes ammonioxydans sp. nov. strain HO-1, oxidizing ammonia to hydroxylamine and ending in the production of N2 gas. Genome analysis revealed that strain HO-1 encoded a complete denitrification pathway but lacks any genes coding for homologous to known ammonia monooxygenases or hydroxylamine oxidoreductases. Our results demonstrated strain HO-1 denitrified nitrite (not nitrate) to N2 and N2 O at anaerobic and aerobic conditions respectively. Further experiments demonstrated that inhibition of aerobic denitrification did not stop ammonia oxidation and N2 production. A gene cluster (dnfT1RT2ABCD) was cloned from strain HO-1 and enabled E. coli accumulated hydroxylamine. Sub-cloning showed that genetic cluster dnfAB or dnfABC already enabled E. coli cells to produce hydroxylamine and further to 15 N2 from (15 NH4 )2 SO4 . Transcriptome analysis revealed these three genes dnfA, dnfB and dnfC were significantly upregulated in response to ammonia stimulation. Taken together, we concluded that strain HO-1 has a novel dnf genetic cluster for ammonia oxidation and this dnf genetic cluster encoded a previously unknown pathway of direct ammonia oxidation (Dirammox) to N2 .

Pseudomonas sp. AOB-7 utilizes PHA granules as a sustained-release carbon source and biofilm carrier for aerobic denitrification of aquaculture water.

Nitrate accumulation causes long-time threat to aquatic animals in recirculating aquaculture system (RAS); thus, nitrate removal is also required in RASs. However, the lack of carbon sources makes denitrification difficult to function. Nitrate removal performance of an aerobic denitrifying and extracellular polyhydroxyalkanoate depolymerase-producing bacterium, Pseudomonas sp. AOB-7, using polyhydroxyalkanoate (PHA) granules as a solid sustained-release carbon source in RAS was evaluated. With the initial nitrate-N concentration of 140 mg/L, the high denitrification rates of 0.056 g NO3--N L-1 day-1 and 0.035 g NO3--N L-1 day-1 were achieved in denitrification medium containing poly-ß-hydroxybutyrate (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), respectively. Significant erosions and pits formed on the surface of the granules made them a good biofilm carrier for AOB-7, and 3-hydroxybutyrate (3-HB) monomer was the major product released to aquatic phase, which was benefit to animals. SEM photos showed that AOB-7 entered and attached on the inside of the PHA particle holes. A 4-week application trial was conducted to reveal the effects of PHB (AOB-7) denitrifying agent and 3-HB produced on growth of zebrafish (Brachydanio rerio) by adding 0.1% (w/v) PHB (AOB-7) denitrifying agent. Result indicated that PHB (AOB-7) denitrifying agent can significantly reduce nitrate-N content in RASs. Compared with the control group, feed coefficient ratio reduced by 18% and weight gain ratio increased by 29% in the PHB (AOB-7) denitrifying agent group. 3-HB monomer produced during the denitrification was speculated to function as a prebiotic and promote zebrafish growth. KEY POINTS: • AOB-7 showed a good aerobic denitrifying ability on PHA granules as sustained-release C source. • PHB (AOB-7) denitrifying agent can significantly reduce nitrate content in RAS. • R-3-HB monomer was the major product released to aquatic phase and function as a prebiotic.

Characterization and mechanism of anti-Aeromonas salmonicida activity of a marine probiotic strain, Bacillus velezensis V4.

The bacterium Aeromonas salmonicida is the causative agent of furunculosis, a systemic, ubiquitous disease of fish in the salmon family, characterized by high mortality and morbidity. Probiotics are a promising approach for prevention of furunculosis in aquaculture. A bacterial strain with anti-A. salmonicida properties, Bacillus velezensis V4, was isolated and the mechanisms underlying these properties were investigated. Anti-A. salmonicida compounds present in cell-free supernatant of V4 were purified and structurally identified as members of the iturin, macrolactin, and difficidin groups. The compounds contributed jointly to inhibition of A. salmonicida, and the diversity of the compounds was related to the versatility of their mode of action. Addition of the compounds to A. salmonicida cell suspensions reduced cell density. Analyses by confocal microscopy and scanning electron microscopy revealed cell membrane disruption, deletion of cellular content, and cell lysis of A. salmonicida. The V4 genome was sequenced, and gene clusters involved in synthesis of anti-Aeromonas compounds were detected and identified. A possible probiotic effect on growth performance of Oncorhynchus mykiss (rainbow trout) was investigated by addition of 0, 1, and 3 % (v/w) V4. Relative to control, mortality was reduced 27.25 % in the 1 % addition group and 81.86 % in the 3 % addition group. Feed coefficient ratio was reduced 19.49 % and weight gain ratio was increased 71.22 % in the 1 % addition group. Our findings demonstrate that V4 is an effective probiotic strain in O. mykiss and has clear potential for both control of furunculosis and growth promotion of aquaculture animals.

Postpartum uterine involution promoted by penetrating-moxibustion therapy: a randomized controlled trial. / é€ç¸æ³•ä¿ƒè¿›äº§åŽå­å®«å¤æ—§ï¼šéšæœºå¯¹ç §è¯•éªŒ.

OBJECTIVES: To observe the effect of penetrating-moxibustion therapy on postpartum uterine involution. METHODS: Eighty puerpera were randomized into an observation group and a control group, 40 cases in each one. In the control group, oxytocin injection was administered by intravenous drip, 20 U each time, once daily. In the observation group, on the base of the treatment as the control group, the penetrating-moxibustion therapy was used at Shenque (GV 8), Qihai (CV 6) and Guanyuan (CV 4), 30 min to 40 min each time, twice a day. The intervention of each group started from the first day after childbirth and lasted 3 days. The uterine volume before and after treatment, and in 42 days of postpartum, the height decrease of the fundus of the uterus, the score of visual analogue scale (VAS) for uterine contraction, the volume of lochia rubra in 1 to 3 days of treatment, and lochia duration were compared between the two groups; and the clinical effect was evaluated. RESULTS: The uterine volume in the observation group was smaller than that of the control group after treatment (P<0.01). In 1 to 3 days of treatment, the height decrease of the fundus of the uterus in the observation group was larger (P<0.01), VAS scores of uterine contraction were lower (P<0.05, P<0.01), the lochia rubra volume was less (P<0.01) than those in the control group. The duration of lochia rubra and lochia was shorter (P<0.01) in the observation group when compared with that of the control group. The favorable rate of uterine involution in the observation group was 95.0% (38/40), higher than that of the control group (75.0%, 30/40, P<0.05). CONCLUSIONS: Penetrating-moxibustion therapy accelerates the recovery of the uterine volume, relieves uterine contraction, shortens the duration of lochia, reduces the lochia volume and promotes the postpartum uterine involution.

[Clinical effect of acupuncture based on the specific response of jing-well point for intractable insomnia: a randomized controlled trial].

OBJECTIVE: To evaluate clinical effect and safety on the basis of detecting the specific response of jing-well point in treatment of intractable insomnia with acupuncture by meridian differentiation. METHODS: Sixty-four patients with intractable insomnia were randomized into an observation group (32 cases, 1 case dropped out and 1 case was eliminated) and a control group (32 cases, 1 case was eliminated). In the observation group, the meridian imbalance value detected at the jing-well point was taken as the evidence so that the corresponding yuan-source and back-shu points were stimulated with acupuncture. In the control group, the routine acupuncture was operated at Baihui (GV 20), Sishencong (EX-HN 1), and bilateral Shenmen (HT 7), Sanyinjiao (SP 6), Shenmai (BL 62) and Zhaohai (KI 6). Besides, the detection at jing-well point was performed for blindness in the control group. In the two groups, the interventions were delivered once daily, 5 times a weeks and for consecutive 4 weeks. In the two groups, the scores of Pittsburgh sleep quality index (PSQI), insomnia severity index (ISI) and the TCM symptom scale were observed before treatment and after 2 and 4 weeks of treatment; the clinical effect and safety were evaluated after treatment; the changes of meridian imbalance value were observed before and after treatment and the correlation analysis with the total score of PSQI was conducted. RESULTS: After 2 and 4 weeks of treatment, except the scores for hypnotic drug in the two groups and sleep disorder after 2 weeks of treatment in the control group, the scores of the other factors and the total scores of PSQI were all reduced when compared with those before treatment in the two groups (P<0.05). After 4 weeks of treatment, except the scores for hypnotic drug in the two groups and sleep disorder in the control group, the scores of the other factors and the total scores of PSQI were lower than those after 2 weeks of treatment in the two groups (P<0.05). After 2 weeks of treatment, the scores for time to fall asleep, sleep efficiency and daytime dysfunction in the observation group were lower than those of the control group (P<0.05); and after 4 weeks of treatment, except the scores for sleep disorder and hypnotic drug, the scores of the other factors and the total score of PSQI in the observation group were all lower than those of the control group (P<0.05). After 2 and 4 weeks of treatment, ISI scores and the scores of TCM symptom scale decreased when compared with those before treatment (P<0.05), and the scores of these two scales after 4 weeks of treatment were lower than those after 2 weeks of treatment (P<0.05) in the two groups; and the scores in the observation group were lower than thoese in the control group (P<0.05). The total effective rate was 93.3% (28/30) in the observation group, higher than that (90.3% [28/31]) in the control group (P<0.05). Of 64 cases, there was only 1 case of mild hematoma in the control group; and no any other adverse events occurred. Among 64 cases, the meridians, with the imbalance frequency ≥30 times, included the pericardium meridian of hand-jueyin and the heart meridian of hand-shaoyin; those with the imbalance frequency ≥20 times, were the kidney meridian of foot-shaoyin, the triple energizers meridian of hand-shaoyang, the gallbladder meridian of foot-shaoyang, the spleen meridian of foot-taiyin and the stomach meridian of foot-yangming. Except the lung meridian of hand-taiyin in the control group, the imbalance value of each meridian was reduced after treatment (P<0.05, P<0.001, P<0.01), and the meridian imbalance value presented a linear positive correlation with the total score of PSQI in the two groups . CONCLUSION: Meridian differentiation acupuncture based on detecting the specific response of jing-well point can significantly improve the sleep quality and reduce the related symptoms in the patients with intractable insomnia. This therapy promotes the conversion of the meridians from the imbalance to the balance and is satisfactory in its safe operation.

Curative effect of acupuncture and moxibustion on insomnia: a randomized clinical trial.

OBJECTIVE: To observe the effect of acupuncture and moxibustion on insomnia and explore its mechanism. METHODS: One hundred and twenty patients were randomly divided into an experiment group and a control group. Sixty patients in the experiment group were treated once a day with acupuncture at Baihui (GV 20), Sishencong (EX-HN 1), Shenmai (BL 62), and Zhaohai (KI 6) and with moxibustion at Baihui (GV 20) and Sishencong (EX-HN 1). Sixty patients in the control group were acupunctured once a day at Shenmen (HT 7), Neiguan (PC 6), and Sanyinjiao (SP 6).The Pittsburgh Sleep Quality Index (PSQI) was used to compare sleep improvement between the two groups. RESULTS: The total effective rate was 87.7% in the experiment group and 76.3% in the control group. The PSQI scores and the total score were lower after treatment than before treatment in both groups. However, the reduction in the experiment group was greater than that in the control group in sleeping quality, time to fall asleep, sleeping disorder, and daytime function (P < 0.05). CONCLUSION: Acupuncture and moxibustion at Baihui (GV 20), Sishencong (EX-HN 1), Shenmai (BL 62), and Zhaohai (KI 6) significantly improved insomnia symptoms in the experiment group compared with the control group.

Copper and cuproptosis: new therapeutic approaches for Alzheimer's disease.

Copper (Cu) plays a crucial role as a trace element in various physiological processes in humans. Nonetheless, free copper ions accumulate in the brain over time, resulting in a range of pathological changes. Compelling evidence indicates that excessive free copper deposition contributes to cognitive decline in individuals with Alzheimer's disease (AD). Free copper levels in the serum and brain of AD patients are notably elevated, leading to reduced antioxidant defenses and mitochondrial dysfunction. Moreover, free copper accumulation triggers a specific form of cell death, namely copper-dependent cell death (cuproptosis). This article aimed to review the correlation between copper dysregulation and the pathogenesis of AD, along with the primary pathways regulating copper homoeostasis and copper-induced death in AD. Additionally, the efficacy and safety of natural and synthetic agents, including copper chelators, lipid peroxidation inhibitors, and antioxidants, were examined. These treatments can restore copper equilibrium and prevent copper-induced cell death in AD cases. Another aim of this review was to highlight the significance of copper dysregulation and promote the development of pharmaceutical interventions to address it.

Acupuncture at Back-Shu point improves insomnia by reducing inflammation and inhibiting the ERK/NF-κB signaling pathway.

BACKGROUND: Insomnia is a disease where individuals cannot maintain a steady and stable sleep state or fail to fall asleep. Western medicine mainly uses sedatives and hypnotic drugs to treat insomnia, and long-term use is prone to drug resistance and other adverse reactions. Acupuncture has a good curative effect and unique advantages in the treatment of insomnia. AIM: To explore the molecular mechanism of acupuncture at Back-Shu point for the treatment of insomnia. METHODS: We first prepared a rat model of insomnia, and then carried out acupuncture for 7 consecutive days. After treatment, the sleep time and general behavior of the rats were determined. The Morris water maze test was used to assess the learning ability and spatial memory ability of the rats. The expression levels of inflammatory cytokines in serum and the hippocampus were detected by ELISA. qRT-PCR was used to detect the mRNA expression changes in the ERK/NF-κB signaling pathway. Western blot and immunohistochemistry were carried out to evaluate the protein expression levels of RAF-1, MEK-2, ERK1/2 and NF-κB. RESULTS: Acupuncture can prolong sleep duration, and improve mental state, activity, diet volume, learning ability and spatial memory. In addition, acupuncture increased the release of 1L-1ß, 1L-6 and TNF-α in serum and the hippocampus and inhibited the mRNA and protein expression of the ERK/NF-κB signaling pathway. CONCLUSION: These findings suggest that acupuncture at Back-Shu point can inhibit the ERK/NF-κB signaling pathway and treat insomnia by increasing the release of inflammatory cytokines in the hippo-campus.

Relative specificity of acupuncture reinforcing and reducing methods.

OBJECTIVE: To probe the effects of acupuncture reinforcing and reducing methods on the human body. METHODS: Studied acupuncture reinforcing and reducing actions in the ancient literature, based on modern clinical and experimental studies, in combination with experience of acupuncture teaching and clinical treatment. RESULTS: The key to the generation of reinforcing and reducing actions is not the acupuncture reinforcing and reducing methods themselves, but instead the functional state of the patient during the acupuncture. When reinforcing and reducing methods act on the human body, a reinforcing-reducing effect is produced through regulation of Qi, indicating that the production of the reinforcing-reducing effect requires a certain condition. Specifically, acupuncture does not produce the reinforcing-reducing effect under all conditions, but can produce a reinforcing effect in patients with deficiency syndrome and a reducing effect in patients with excess syndrome. CONCLUSION: Reinforcing and reducing methods each have therapeutic effects on both deficiency syndrome and excess syndrome, but a reinforcing method should be selected first for deficiency syndrome and a reducing method should be selected first for excess syndrome.

Degradation of amoxicillin by newly isolated Bosea sp. Ads-6.

Amoxicillin (AMX), one of the micro-amount hazardous pollutants, was frequently detected in environments, and of great risks to environments and human health. Microbial degradation is a promising method to eliminate pollutants. In this study, an efficient AMX-degrading strain, Ads-6, was isolated and characterized. Strain Ads-6, belonging to the genus Bosea, was also able to grow on AMX as the sole carbon and nitrogen source, with a removal of ~60% TOC. Ads-6 exhibited strong AMX-degrading ability at initial concentrations of 0.5-2 mM and pH 6-8. Addition of yeast extract could significantly enhance its degrading ability. Many degradation intermediates were identified by HPLC-MS, including new ones such as two phosphorylated products which were firstly defined in AMX degradation. A new AMX degradation pathway was proposed accordingly. Moreover, the results of comparative transcriptomes and proteomes revealed that ß-lactamase, L, D-transpeptidase or its homologous enzymes were responsible for the initial degradation of AMX. Protocatechuate branch of the beta-ketoadipate pathway was confirmed as the downstream degradation pathway. These results in the study suggested that Ads-6 is great potential in biodegradation of antibiotics as well as in the bioremediation of contaminated environments.

Alcaligenes ammonioxydans HO-1 antagonizes Bacillus velezensis via hydroxylamine-triggered population response.

Antagonism is a common behavior seen between microbes in nature. Alcaligenes ammonioxydans HO-1 converts ammonia to nitrogen under aerobic conditions, which leads to the accumulation of extracellular hydroxylamine (HA), providing pronounced growth advantages against many bacterial genera, including Bacillus velezensis V4. In contrast, a mutant variant of A. ammonioxydans, strain 2-29, that cannot produce HA fails to antagonize other bacteria. In this article, we demonstrate that cell-free supernatants derived from the antagonistic HO-1 strain were sufficient to reproduce the antagonistic behavior and the efficiency of this inhibition correlated strongly with the HA content of the supernatant. Furthermore, reintroducing the capacity to produce HA to the 2-29 strain or supplementing bacterial co-cultures with HA restored antagonistic behavior. The HA-mediated antagonism was dose-dependent and affected by the temperature, but not by pH. HA caused a decline in biomass, cell aggregation, and hydrolysis of the cell wall in exponentially growing B. velezensis bulk cultures. Analysis of differential gene expression identified a series of genes modulating multicellular behavior in B. velezensis. Genes involved in motility, chemotaxis, sporulation, polypeptide synthesis, and non-ribosomal peptide synthesis were all significantly downregulated in the presence of HA, whereas autolysis-related genes showed upregulation. Taken together, these findings indicate that HA affects the population response of coexisting strains and also suggest that A. ammonioxydans HO-1 antagonize other bacteria by producing extracellular HA that, in turn, acts as a signaling molecule.

Denitrification characteristics of a marine origin psychrophilic aerobic denitrifying bacterium.

A psychrophilic aerobic denitrifying bacterium, strain S1-1, was isolated from a biological aerated filter conducted for treatment of recirculating water in a marine aquaculture system. Strain S1-1 was preliminarily identified as Psychrobacter sp. based on the analysis of its 16S rRNA gene sequence, which showed 100% sequence similarity to that of Psychrobacter sp. TSBY-70. Strain S1-1 grew well either in high nitrate or high nitrite conditions with a removal of 100% nitrate or 63.50% nitrite, and the total nitrogen removal rates could reach to 46.48% and 31.89%, respectively. The results indicated that nitrate was mainly reduced in its logarithmic growth phase with a very low level accumulation of nitrite, suggesting that the aerobic denitrification process of strain S1-1 occurred mainly in this phase. The GC-MS results showed that N2O was formed as the major intermediate during the aerobic denitrifying process of strain S1-1. Finally, factors affecting the growth of strain S1-1 and its aerobic denitrifying ability were also investigated. Results showed that the optimum aerobic denitrification conditions for strain S1-1 were sodium succinate as carbon source, C/N ratio15, salinity 10 g/L NaCl, incubation temperature 20 degrees C and initial pH 6.5.

[Design and application of Baihui moxibustion box].

On the base of the principle of penetrating moxibustion and in combination with free adjustment devices such as movable U-shaped moxa stick holder and movable clamp, a new type of moxibustion box exerted on the head is designed, with precise positioning and sufficient heat intensity. Baihui moxibustion box is composed of two sections, i.e. body section and pillow section, which is as one structure. There are several vertical bar-shaped holes distributed evenly on the movable door outside moxa box. The U-shaped moxa stick holder on the inner side of the bar-shaped hole is connected with the fixed clamp on the outside, which is movable up and down, forward and backward for height adjustment. Such moxibustion box is characterized as accurate positioning, energy saving, temperature control and manpower saving.

[Advances in bioremediation of polycyclic aromatic hydrocarbons contaminated soil].

Polycyclic aromatic hydrocarbons (PAHs) are a class of persistent pollutants that are widely distributed in the environment. Due to their stable structure and poor degradability, PAHs exhibit carcinogenic, teratogenic, and mutagenic toxicity to the ecological environment and organisms, thus increasing attentions have been paid to their removals and remediation. Green, safe and economical technologies are widely used in the bioremediation of PAHs-contaminated soil. This article summarizes the present status of PAHs pollution in soil of China from the aspects of origin, migration, fate, and pollution level. Meanwhile, the types of microorganisms and plants capable of degrading PAHs, as well as the underlying mechanisms, are summarized. The features of three major bioremediation technologies, i.e., microbial remediation, phytoremediation, and joint remediation, are compared. Analysis of the interaction mechanisms between plants and microorganisms, selection and cultivation of stress-resistant strains and plants, as well as safety and efficacy evaluation of practical applications, are expected to become future directions in this field.

[Adjuvant treatment of penetrating moxibustion at governor vessel for persistent allergic rhinitis of deficiency-cold syndrome].

OBJECTIVE: To explore the efficacy and action mechanism of penetrating moxibustion at governor vessel for persistent allergic rhinitis of deficiency-cold syndrome. METHODS: Ninety patients with persistent allergic rhinitis of deficiency-cold syndrome were randomly divided into an observation group (n = 45) and a control group (n = 45). The patients in the control group were treated with momethasone furoate nasal spray, 2 sprays per side per time, once a day. On the basis of treatment in the control group, the patients in the observation group were additionally treated with penetrating moxibustion at governor vessel, 2 h per treatment, once a week. Both groups were treated for 4 weeks. The TCM symptom score, visual analogue scale (VAS) score and rhinoconjunctivitis quality of life questionnaire (RQLQ) score were observed in the two groups before and after treatment. The serum level of immunoglobulin E (IgE) and complete blood count of eosinophil (EOS) were measured before and after treatment, and the clinical effects were compared. RESULTS: Compared before treatment, the TCM symptom scores, VAS scores, RQLQ scores, serum levels of IgE and complete blood count of EOS in the two groups were all reduced after treatment (P<0.05), and those in the observation group were lower than those in the control group (P<0.05). The total effective rate in the observation group was 95.6% (43/45), which was higher than 82.2% (37/45) in the control group (P<0.05). CONCLUSION: Based on the momethasone furoate nasal spray, the adjuvant treatment of penetrating moxibustion at governor vessel could significantly improve the clinical symptoms in patients with persistent allergic rhinitis of deficiency-cold syndrome, and its mechanism may be related to the regulation of immune disorder.

Effect and Safety of Penetrating Moxibustion in Treatment of Migraine without Aura: A Randomized Controlled Trial.

OBJECTIVE: To observe the clinical effect of penetrating moxibustion on migraine without aura (MO) patients. METHODS: Totally 60 MO patients from the Acupuncture Clinic of the Third Affiliated Hospital of Henan University of Chinese Medicine were collected from November 2015 to February 2017. All patients were assigned to a treatment group and a control group using a random number table, 30 cases in each group. The treatment group was treated with penetrating moxibustion, and the control group was treated with mild moxibustion, thrice a week for 4 consecutive weeks. The total effective rate, Visual Analogue Scale (VAS) scores, headache intensity, and Migraine Specific Quality of Life Questionnaire (MSQ) scores of patients after treatment were compared between the two groups. The moxibustion sensation and reaction after moxibustion were observed, and the adverse reactions were evaluated. All patients were followed up at 4 and 16 weeks after treatment. RESULTS: The total effective rate of the treatment group was significantly higher than that of the control group (93.33% vs. 80.00%, P<0.05). The improvement of VAS scores, headache intensity, and the role restrictive and role preventive scores in MSQ in the treatment group was better than those in the control group (P<0.05). The person-time of moxibustion sensations of itching, numbness and cold as well as flushing and sweating after moxibustion in the treatment group was all significantly higher than that in the control group (P<0.01). There was no significant difference in safety evaluation between the two groups (P>0.05). CONCLUSIONS: Penetrating moxibustion can significantly relieve pain and improve quality of life of MO patients. After penetrating moxibustion, flushing and sweating of patients were obvious, and the curative effect was superior to the mild moxibustion.

Determination of intermediates and mechanism for soot combustion with NOx/O2 on potassium-supported Mg-Al hydrotalcite mixed oxides by in situ FTIR.

The soot combustion with NO(x) and/or O(2) on potassium-supported Mg-Al hydrotalcite mixed oxides under tight contact condition was studied using temperature-programmed oxidation (TPO), isothermal reaction and in situ FTIR techniques. The presence of NO(x) in O(2) favors the soot combustion at lower temperatures (<300 °C). However, a little suppression was observed at higher temperatures (>300 °C), which was accompanied by a substantial NO(x) reduction. The ketene (C═C═O) and isocyanate (NCO(-)) species were determined as the reaction intermediates. In NO(x) + O(2), NO(2) directly interacts with the free carbon sites (C═C*) through two parallel reactions: (1) NO(2) + C═C* → C═C═O + NO; (2) NO(2) + C═C* → NCO(-) + CO(2). The two reactions can proceed easily, which accounts for the promotion effect of NO(x) on soot combustion at lower temperatures. The further oxidation of NCO(-) by NO(2) or O(2) is responsible for the simultaneous reduction of NO(x). However, the reactions between NO(2) and C═C* are limited by the amount of free carbon sites, which can be provided by the oxidation of soot by O(2) at higher temperatures. The interaction of NO(x) and catalyst results in the formation of nitrates and nitrites, which poisoned the active K sites.

[Identification and characteristics of a marine aerobic denitrifying bacterium].

OBJECTIVE: An aerobic denitrifying bacterium, strain 2-8, was isolated from a biological aerated filter in a recirculating marine aquaculture system, phylogeny and characteristics of the strain was further studied. METHODS: Sequence of the 16S rRNA gene was analyzed and the factors affect the denitrifying ability of strain 2-8 were investigated, including carbon source, C/N ratio, initial pH, NaCl concentration, temperature and shaker speed. RESULTS: Strain 2-8 was identified as Pseudomonas sp. based on the analysis of its 16S rRNA gene sequence which showed highest similarity (99.9%) to Pseudomonas segetis FR1439(T) (AY770691). The results indicated that carbon source and C/N ratio exhibited significant influences on aerobic denitrifying capacity of strain 2-8. Strain 2-8 could grow a little on acetate, succinate and citrate as sole carbon source, and the removal rates of NO3- -N at 140 mg/L were higher than 65% despite of the accumulation of NO2- -N around 35 mg/L. It grew quite well on glucose as sole carbon source, however, the removal rate of nitrate was not so high as on other carbon source. The optimum C/N ratio was 15, as lower C/N ratios may lead to nitrite accumulation. The optimum temperature and pH for its aerobic denitrification were 30 degrees C and 7.5, respectively. Strain 2-8 could grow and exhibit aerobic denitrifying ability at a wide range of NaCl concentrations (0-30 g/L). The highest nitrogen removal appeared under the condition of 160 r/min shaking culture. CONCLUSION: When cultured in the conditions of NO3- -N at 140 mg/L, sodium citrate as sole carbon source, C/N ratio at 15, pH 7.5, NaCl at 30 g/L, 30 degrees C and 160 r/min of the shaker, strain 2-8 removed up to 92% of the nitrogen within 48 hours, and no nitrite accumulation.

[Exploration and practice of micro-video teaching of Characteristic Clinical Technology of Acupuncture and Moxibustion].

In this paper, the micro-video teaching mode was explored in the course construction of Characteristic Clinical Technology of Acupuncture and Moxibustion. The micro-video teaching contents include the academic thought, experience in diagnosis and treatment, characteristic technology and clinical manipulation of famous acupuncture experts in the Henan University of CM. Each micro-video film is designed within 15-18 min, including three sections of knowledge, i.e. basic theory, technological application and clinical manipulation. Each section is designed within 5-6 min. The construction of the teaching course of Characteristic Clinical Technology of Acupuncture and Moxibustion is the innovation of practice mode of TCM and the new approach to the inheritance of the experience of experts. The construction of micro-video teaching course propels the reform of teaching mode, improves the learning initiative of students and clinical manipulative ability so as to improve the teaching effect and quality.