Precursor Quantitation Methods for Next Generation Food Production.

Food is essential for human survival. Nowadays, traditional agriculture faces challenges in balancing the need of sustainable environmental development and the rising food demand caused by an increasing population. In addition, in the emerging of consumers' awareness of health related issues bring a growing trend towards novel nature-based food additives. Synthetic biology, using engineered microbial cell factories for production of various molecules, shows great advantages for generating food alternatives and additives, which not only relieve the pressure laid on tradition agriculture, but also create a new stage in healthy and sustainable food supplement. The biosynthesis of food components (protein, fats, carbohydrates or vitamins) in engineered microbial cells often involves cellular central metabolic pathways, where common precursors are processed into different proteins and products. Quantitation of the precursors provides information of the metabolic flux and intracellular metabolic state, giving guidance for precise pathway engineering. In this review, we summarized the quantitation methods for most cellular biosynthetic precursors, including energy molecules and co-factors involved in redox-reactions. It will also be useful for studies worked on pathway engineering of other microbial-derived metabolites. Finally, advantages and limitations of each method are discussed.

The effects of Jin's three-needle acupuncture therapy on EEG alpha rhythm of stroke patients.

BACKGROUND: In China, Jin's three-needle acupuncture therapy has been widely used for stroke treatment. However, its mechanism remains unclear. The aim of this study was to investigate the effects of Jin's three-needle acupuncture therapy on stroke patients using multi-channel EEGs. MATERIALS AND METHODS: Fifty stroke patients participated and their EEG signals were recorded before, during, and after acupuncture. RESULTS: Comparing with baseline before acupuncture, the alpha rhythm was significantly increased during the acupuncture needle retention stage and became stronger after removing the needles. The increase of alpha rhythm occurred when inserting and removing the needles, with its high amplitude remaining during retention and after removal of the needles. As the alpha rhythm was significantly correlated with patients' movement and daily-living abilities, the alteration of alpha rhythm during and after acupuncture suggests a possible electrophysiological mechanism of the curative effect of acupuncture for stroke rehabilitation. CONCLUSION: From the electrophysiological study, Jin's three-needle acupuncture therapy is efficient in stroke patients.

Effect of Taichong (LR 3) acupuncture in spontaneously hypertensive rats.

OBJECTIVE: To evaluate the effects of Taichong (LR 3) acupuncture points (acupoints) on the expression of glucose transporter protein 1 (GLUT1) in the hypothalamus of spontaneously hypertensive rats (SHRs) as measured by combined positron emission tomography and computed tomography (PET-CT). METHODS: Spontaneously hypertensive rats (SHR) were divided into model, Taichong (LR 3) acupuncture, and sham groups. Additionally, Tokyo Wistar rats were used as the control group. Changes in blood pressure were recorded in different groups of rats before and after the corresponding treatment. Hematoxylin and eosin (HE) staining was used to study basic morphological changes, and immunohistochemistry was used to determine GLUT1 expression in the hypothalamus. Further, PET-CT was utilized to elucidate the antihypertensive mechanism after acupuncture at the Taichong (LR 3) acupoints. RESULTS: PET-CT indicated activation of the hypothalamus. Measurement of blood pressure showed that acupuncture at the Taichong (LR 3) acupoints lowered blood pressure. HE staining did not show any significant pathological changes, although differences in cell number were observed. Immunohistochemical analysis indicated a GLUT1 downregulation in the SHRs of the Taichong (LR 3) acupuncture group after the treatment. CONCLUSION: Acupuncture at Taichong (LR 3) acupoints lowered blood pressure in SHRs, with possible mechanisms being changes in cell number and GLUT1 expression in the hypothalamus.

[Using Machine Learning to Investigate Factors Influencing the Efficacy of "Shoulder Tri-needles Therapy" in Treatment of Shoulder-hand Syndrome in 586 Stroke Patients].

OBJECTIVE: To analyze the factors influencing the therapeutic effect of "Shoulder Tri-needles therapy" in the treatment of shoulder-hand syndrome of stroke patients by using machine learning approach, so as to provide a feasibility for improving clinical efficacy. METHODS: A total of 586 stroke patients with shoulder-hand syndrome eligible for this study were involved in our machine learning experiments for classification of the influential factors. Their data including the age, gender, pulse condition, complexion, tongue quality, tongue coating, disease stage, body mass index (BMI), blood pressure, blood glucose, blood triglyceride, blood total cholesterol, smoking history, drinking history, and final outcomes were extracted from the medical record system (from Oct. of 2014 to Jan. of 2017 in the First Affiliated Hospital and Shenzhen Futian Hospital of Guangzhou University of Chinese Medicine). The single rule algorithm (1 R) was adopted to learn, followed by optimization with Repeated Incremental Pruning to Produce Error Reduction (RIPPER) algorithm, and C 5.0 decision tree algorithm. RESULTS: The accurate classification rates of 1 R, RIPPER and decision tree model were 87.37%(512/586), 95.90% (562/586), and 97.10% (569/586), respectively. The final outcomes of machine learning of this study showed that the disease stage (acute or recovery stage), complexion difference, tongue coating difference, blood pressure level, consumption of alcohol, BMI, and smoking habit were the most important factors influencing the therapeutic effect of "Shoulder Tri-needles" in the treatment of shoulder-hand syndrome of stroke patients. CONCLUSION: The disease stage, complexion and tongue identification, blood pressure level, alcohol drinking and smoking habits, and BMI are the principal factors affecting the therapeutic effect of "Shoulder Tri-needles therapy" in the treatment of shoulder-hand syndrome of stroke patients.

[A Meta-analysis of Effectiveness of Acupoint Catgut Embedding Therapy for Insomnia].

OBJECTIVE: To evaluate the effectiveness and superiority of acupoint catgut-embedding in the treatment of insomnia. METHODS: Randomized controlled trials (RCTs) for insomnia treated by acupoint implantation of catgut were included in the present study. We searched the related articles from 6 databases (PubMed, Google Scholar, CBM, CNKI, VIP and Wanfang) from the date of inception to March 19, 2017 by using keywords "catgut implantation at acupuncture point" or "catgut implantation at acupoint" and "insomnia" or "agrypnia" and "randomised (or randomized) clinical trial". Quality assessments were performed by two researchers independently using GRADE profile 3.6, followed by crosscheck. Data were analyzed using software Revman 5.3 and STATA 14.0. RESULTS: Twenty-two trials met the inclusion criteria were included. They recruited 1 714 cases of insomnia. GRADE grading suggests a low quality of these articles. Heterogeneous test (numerical test) showed χ2＝31.11, P＝0.07, I2＝32%. Gabriz chart and sensitivity analysis ultimately determined that these trials have smaller or no heterogenity. The effective rate of acupoint catgut embedding for insomnia was higher than that of the control group (OR＝ 3.77; 95% CI: 2.63－5.41; P<0.00001). CONCLUSION: Acupoint catgut embedding therapy has a positive role in clinical treatment of insomnia. But due to lower quality of current literature, more high-quality randomized controlled clinical trials are required.

[A Meta-analysis of the Effectiveness of Acupuncture in the Treatment of Epilepsy].

OBJECTIVE: To evaluate the efficacy and safety of acupuncture in the treatment of epilepsy. METHODS: We first collected research data about randomized control trials (RCTs) of acupuncture treatment of epilepsy from databases CNKI, CBM, VIP and Wanfang using key words "(acupuncture)", "(epilepsy)"or "(epilepsy)"or "(epilepsy)", and from PubMed, Google Scholar, and Cochrane Library using key words "acupuncture" "needling" "prod" "epilepsy" "epilepsies" or "seizure disorder" or "simple seizure" from the date of database construction to May 5, 2017. Then, two researchers extracted the study outcomes and evaluated the evidence qualification of the research data independently using GRADE profile 3.6 software and analyzed the primary outcome indexes using RevMan 5.3 and STATA 14.0. RESULTS: Eleven articles containing 827 cases were included in the Meta-analysis. We found that acupuncture or acupuncture in combination with other conventional drugs had a significantly better efficacy than conventional drugs in the treatment of epilepsy [OR=3.94, 95%CI (2.49, 6.24), Z=5.85, P<0.000 01]. There was a statistical heterogeneity (P=0.04, l2=60%), and the Meta-analysis was not feasible for the studies of acupuncture vs medication. Combination of acupuncture and Chinese traditional drugs was significantly better than simple Chinese traditional drugs [OR = 4.61, 95%CI (2.18, 9.74), Z=4.01, P<0.000 1], and combination of acupuncture and wes-tern medicines was obviously better than simple western medicines [OR=3.07, 95%CI(1.24, 7.65), Z=2.41, P=0.02] in the treatment of epilepsy. CONCLUSION: Acupuncture therapy may have a positive effect in the treatment of epilepsy but the conclusion needs further verification due to very fewer high-quality and well-designed RCTs found at the present.

[Influencing factors of the occurrence of propagated sensation effects at Zusanli (ST 36) based on artificial intelligence technology].

OBJECTIVE: To explore the influencing factors of the occurrence of propagated sensation effects at Zusanli (ST 36) based on artificial intelligence technology. METHODS: A total of 272 undergraduates and postgraduates of Guangzhou University of TCM were recruited. The basic information, including TCM constitution, body weight index (BMI), pulse, tongue and coating, face complexion, as well as the occurrence of propagated sensation effects at Zusanli (ST 36) with different acupuncture needles and techniques, were recorded. The repeated incremental pruning to produce error reduction (RIPPER) algorithm was applied to predict the results and summarize the rules of relevant factors. RESULTS: ① The propagated sensation effects were often observed in the subject with yin-deficiency constitution (55 cases in total, of them 1 case was misclassified). ② The propagated sensation effects were observed in the subject with balanced yin-yang constitution (39 cases in total, of them 0 case was misclassified). ③ The propagated sensation effects were observed in the subject with phlegm-damp constitution combined with thin tongue and white coating (31 cases in total, of them 8 cases were misclassified). ④ The propagated sensation effects were observed in the subject with phlegm-damp constitution combined with light tongue coating (7 cases in total, of them 2 cases were misclassified). ⑤ The propagated sensation effects were observed in all the subject with special constitution (6 cases in total, of them 0 cases were misclassified). ⑥ The propagated sensation effects were not observed in the subject with other constitution or the combination of constitution and symptoms (134 cases in total, of them 4 cases were misclassified). The accuracy rate of classification was 94.49%. The difference of acupuncture needles and techniques was not significant in propagated sensation effects (P>0.05). CONCLUSION: The TCM constitution may be the most essential factor in the occurrence of propagated sensation effects.

Rational design of a Kv1.3 channel-blocking antibody as a selective immunosuppressant.

A variable region fusion strategy was used to generate an immunosuppressive antibody based on a novel "stalk-knob" structural motif in the ultralong complementary-determining region (CDR) of a bovine antibody. The potent Kv1.3 channel inhibitory peptides Moka1-toxin and Vm24-toxin were grafted into different CDRs of the humanized antibodies BVK and Synagis (Syn) using both ß-sheet and coiled-coil linkers. Structure-activity relationship efforts led to generation of the fusion protein Syn-Vm24-CDR3L, which demonstrated excellent selectivity and potency against effector human memory T cells (subnanomolar to picomolar EC50 values). This fusion antibody also had significantly improved plasma half-life and serum stability in rodents compared with the parent Vm24 peptide. Finally, this fusion protein showed potent in vivo efficacy in the delayed type hypersensitivity in rats. These results illustrate the utility of antibody CDR fusions as a general and effective strategy to generate long-acting functional antibodies, and may lead to a selective immunosuppressive antibody for the treatment of autoimmune diseases.

Microstructure and properties of the Ti/Al2O3/NiCr composites fabricated by explosive compaction/cladding.

Titanium/aluminum oxide/nickel chromium (Ti/Al2O3/NiCr) composite bar prepared by explosive compaction/cladding technique represents a new kind of sandwich-structural composites for medical application. Formation of the interfaces of Ti/Al2O3 and Al2O3/NiCr govern the properties of the composite material. The electrical resistivity and microstructure of the intermediate layer and the interfaces of the Ti/Al2O3/NiCr explosive compaction/cladding bar are investigated by means of four-point probe analysis, optical microscopy, scanning electron microscopy, electron microprobe analysis, and X-ray diffraction. The Ti/Al2O3/NiCr composite bar is characterized by the consolidated ceramic intermediate layer and the metallurgical bonding interfaces. The intermediate ceramic layer plays a role of insulation and thermal conductance in this composite. The average shear strength of the composite bar is about 9.36 MPa. The heat affected zone characterized by relatively larger sizes of grains is distinguished from the other part of the Ti tube. The intermetallics AlTi3 and Al0.9Ni4.22 are generated at the intermediate ceramic layer. Formation mechanism of the interfaces of the explosive compaction/cladding bar are described.