The interaction of trehalose and molten globule state soybean 11S globulin and its impact on foaming capacities.

BACKGROUND: Soybean 11S globulin has good functional properties, which are widely used in the field of food. However, natural soybean 11S globulin (N-11S) has low flexibility and is easy to aggregate, impacting its foaming process. Studies have shown that soybean 11S globulin in molten globule state (MG-11S) has better molecular flexibility than N-11S, and trehalose has been shown to improve the properties of proteins. Therefore, this study investigated the interaction mechanism between trehalose and MG-11S, and its impact on rheological and foaming properties of MG-11S. RESULTS: The molecular docking and intrinsic fluorescence results showed that hydrogen bonding was the main interaction force at lower than 0.5 mol L-1 trehalose added. Meanwhile, rheology and foaming showed that the MG-11S-trehalose complexes had better viscoelasticity, foaming ability (66.67-86.67%) and foaming stability (75.00-89.29%) than N-11S (16.67% foaming ability and 40.00% foaming stability); however, when the trehalose was higher than 0.5 mol L-1 , molecular crowding occurred and H-bonds were weakened, resulting in reduction of foaming capacities. Microstructure determination showed that trehalose attached to the surface of foam membrane; meanwhile, the foaming structure of the complex with 0.5 mol L-1 trehalose had a thicker liquid film with decreased drainage rate, less agglomeration and disproportionation of foam, illustrating the best foaming ability and foaming stability. CONCLUSION: The results suggested that trehalose at different concentrations can interact with MG-11S through different mechanisms, and improve the foaming capacity of MS-11S. This provided a reference for the application of MS-11S in foaming food. © 2022 Society of Chemical Industry.

Evaluating the effectiveness of Hong Kong's border restriction policy in reducing COVID-19 infections.

This study evaluates the effectiveness of Hong Kong's strict border restrictions with mainland China in curbing the transmission of COVID-19. Combining big data from Baidu Population Migration with traditional meteorological data and census data for over 200 Chinese cities, we utilize an advanced quantitative approach, namely synthetic control modeling, to produce a counterfactual "synthetic Hong Kong" without a strict border restriction policy. We then simulate infection trends under the hypothetical scenarios and compare them to actual infection numbers. Our counterfactual synthetic control model demonstrates a lower number of COVID-19 infections than the actual scenario, where strict border restrictions with mainland China were implemented from February 8 to March 6, 2020. Moreover, the second synthetic control model, which assumes a border reopen on 7 May 2020 demonstrates nonpositive effects of extending the border restriction policy on preventing and controlling infections. We conclude that the border restriction policy and its further extension may not be useful in containing the spread of COVID-19 when the virus is already circulating in the local community. Given the substantial economic and social costs, and as precautionary measures against COVID-19 becomes the new normal, countries can consider reopening borders with neighbors who have COVID-19 under control. Governments also need to closely monitor the changing epidemic situations in other countries in order to make prompt and sensible amendments to their border restriction policies.

Spatial congruency or mismatch? Analyzing the COVID-19 potential infection risk and urban density as businesses reopen.

Countries worldwide are reopening their businesses despite the continuing COVID-19 crisis and the emergence of new variants. In this context, knowing whether the reopening of businesses at various locations exposes higher risk to the public is essential. Whether urban density correlates with the potential infection risk as concluded by previous studies of the COVID-19 pandemic remains unknown. In this study, taking the Macau Peninsula as a testbed, we first identified business locations for daily activities according to the latest point of interest (POI) data and generated the potential risk surface for COVID-19 infection. Then, using the cellular phone network and urban footprint data, we further analyzed the spatial relationship between COVID-19 potential risk and urban density of population and morphology through visual analytics. Results show that while some degree of spatial congruency exists between medium-risk peaks and urban density hotspots, apparent spatial mismatch exists for high-risk peaks, indicating that the traditional planning control based on urban density is inadequate for mitigating public health risks. POI-based spatial layout and configuration better reflecting business services and associated human activities are recommended in future planning and policy-making for more resilient cities in the post-pandemic era.

Comprehensive Analysis of CRISPR/Cas9-Mediated Mutagenesis in Arabidopsis thaliana by Genome-wide Sequencing.

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) system has been widely applied in functional genomics research and plant breeding. In contrast to the off-target studies of mammalian cells, there is little evidence for the common occurrence of off-target sites in plants and a great need exists for accurate detection of editing sites. Here, we summarized the precision of CRISPR/Cas9-mediated mutations for 281 targets and found that there is a preference for single nucleotide deletions/insertions and longer deletions starting from 40 nt upstream or ending at 30 nt downstream of the cleavage site, which suggested the candidate sequences for editing sites detection by whole-genome sequencing (WGS). We analyzed the on-/off-target sites of 6 CRISPR/Cas9-mediated Arabidopsis plants by the optimized method. The results showed that the on-target editing frequency ranged from 38.1% to 100%, and one off target at a frequency of 9.8%-97.3% cannot be prevented by increasing the specificity or reducing the expression level of the Cas9 enzyme. These results indicated that designing guide RNA with high specificity may be the preferred factor to avoid the off-target events, and it is necessary to predict or detect off-target sites by WGS-based methods for preventing off targets caused by genome differences in different individuals.

Ultrasensitive Single Fluorescence-Labeled Probe-Mediated Single Universal Primer-Multiplex-Droplet Digital Polymerase Chain Reaction for High-Throughput Genetically Modified Organism Screening.

As genetically modified (GM) technology develops and genetically modified organisms (GMOs) become more available, GMOs face increasing regulations and pressure to adhere to strict labeling guidelines. A singleplex detection method cannot perform the high-throughput analysis necessary for optimal GMO detection. Combining the advantages of multiplex detection and droplet digital polymerase chain reaction (ddPCR), a single universal primer-multiplex-ddPCR (SUP-M-ddPCR) strategy was proposed for accurate broad-spectrum screening and quantification. The SUP increases efficiency of the primers in PCR and plays an important role in establishing a high-throughput, multiplex detection method. Emerging ddPCR technology has been used for accurate quantification of nucleic acid molecules without a standard curve. Using maize as a reference point, four heterologous sequences ( 35S, NOS, NPTII, and PAT) were selected to evaluate the feasibility and applicability of this strategy. Surprisingly, these four genes cover more than 93% of the transgenic maize lines and serve as preliminary screening sequences. All screening probes were labeled with FAM fluorescence, which allows the signals from the samples with GMO content and those without to be easily differentiated. This fiveplex screening method is a new development in GMO screening. Utilizing an optimal amplification assay, the specificity, limit of detection (LOD), and limit of quantitation (LOQ) were validated. The LOD and LOQ of this GMO screening method were 0.1% and 0.01%, respectively, with a relative standard deviation (RSD) < 25%. This method could serve as an important tool for the detection of GM maize from different processed, commercially available products. Further, this screening method could be applied to other fields that require reliable and sensitive detection of DNA targets.

Multiplex enrichment quantitative PCR (ME-qPCR): a high-throughput, highly sensitive detection method for GMO identification.

Among all of the high-throughput detection methods, PCR-based methodologies are regarded as the most cost-efficient and feasible methodologies compared with the next-generation sequencing or ChIP-based methods. However, the PCR-based methods can only achieve multiplex detection up to 15-plex due to limitations imposed by the multiplex primer interactions. The detection throughput cannot meet the demands of high-throughput detection, such as SNP or gene expression analysis. Therefore, in our study, we have developed a new high-throughput PCR-based detection method, multiplex enrichment quantitative PCR (ME-qPCR), which is a combination of qPCR and nested PCR. The GMO content detection results in our study showed that ME-qPCR could achieve high-throughput detection up to 26-plex. Compared to the original qPCR, the Ct values of ME-qPCR were lower for the same group, which showed that ME-qPCR sensitivity is higher than the original qPCR. The absolute limit of detection for ME-qPCR could achieve levels as low as a single copy of the plant genome. Moreover, the specificity results showed that no cross-amplification occurred for irrelevant GMO events. After evaluation of all of the parameters, a practical evaluation was performed with different foods. The more stable amplification results, compared to qPCR, showed that ME-qPCR was suitable for GMO detection in foods. In conclusion, ME-qPCR achieved sensitive, high-throughput GMO detection in complex substrates, such as crops or food samples. In the future, ME-qPCR-based GMO content identification may positively impact SNP analysis or multiplex gene expression of food or agricultural samples. Graphical abstract For the first-step amplification, four primers (A, B, C, and D) have been added into the reaction volume. In this manner, four kinds of amplicons have been generated. All of these four amplicons could be regarded as the target of second-step PCR. For the second-step amplification, three parallels have been taken for the final evaluation. After the second evaluation, the final amplification curves and melting curves have been achieved.

Real-time quantitative nicking endonuclease-mediated isothermal amplification with small molecular beacons.

Techniques of isothermal amplification have recently made great strides, and have generated significant interest in the field of point-of-care detection. Nicking endonuclease-mediated isothermal amplification (NEMA) is an example of simple isothermal technology. In this paper, a real-time quantitative nicking endonuclease-mediated isothermal amplification with small molecular beacons (SMB-NEMA) of improved specificity and sensitivity is described. First, we optimized the prohibition of de novo synthesis by choosing Nt·BstNBI endonuclease. Second, the whole genome was successfully amplified with Nt·BstNBI (6 U), betaine (1 M) and trehalose (60 mM) for the first time. Third, we achieved 10 pg sensitivity for the first time after adding a small molecular beacon that spontaneously undergoes a conformational change when hybridizing to target, and the practical test validated the assay's application. The small molecular beacon has a similar melting temperature to the reaction temperature, but is approximately 10 bp shorter than the length of a traditional molecular beacon. A new threshold regulation was also established for isothermal conditions. Finally, we established a thermodynamic model for designing small molecular beacons. This multistate model is more correct than the traditional algorithm. This theoretical and practical basis will help us to monitor SMB-NEMA in a quantitative way. In summary, our SMB-NEMA method allows the simple, specific and sensitive assessment of isothermal DNA quantification.

A Novel Pretreatment-Free Duplex Chamber Digital PCR Detection System for the Absolute Quantitation of GMO Samples.

Digital polymerase chain reaction (PCR) has developed rapidly since it was first reported in the 1990s. However, pretreatments are often required during preparation for digital PCR, which can increase operation error. The single-plex amplification of both the target and reference genes may cause uncertainties due to the different reaction volumes and the matrix effect. In the current study, a quantitative detection system based on the pretreatment-free duplex chamber digital PCR was developed. The dynamic range, limit of quantitation (LOQ), sensitivity and specificity were evaluated taking the GA21 event as the experimental object. Moreover, to determine the factors that may influence the stability of the duplex system, we evaluated whether the pretreatments, the primary and secondary structures of the probes and the SNP effect influence the detection. The results showed that the LOQ was 0.5% and the sensitivity was 0.1%. We also found that genome digestion and single nucleotide polymorphism (SNP) sites affect the detection results, whereas the unspecific hybridization within different probes had little side effect. This indicated that the detection system was suited for both chamber-based and droplet-based digital PCR. In conclusion, we have provided a simple and flexible way of achieving absolute quantitation for genetically modified organism (GMO) genome samples using commercial digital PCR detection systems.

Development and application of a quantitative loop-mediated isothermal amplification method for detecting genetically modified maize MON863.

BACKGROUND: A SYBR Green I-based quantitative loop-mediated isothermal amplification (LAMP) assay was developed for the rapid detection of genetically modified maize MON863. A set of primers was designed based on the integration region of the Cry3Bb1 and tahsp17 genes. RESULTS: The qualitative and quantitative reaction conditions (dNTPs, betaine, primers, Mg(2+), Bst polymerase, temperature, reaction time) were optimized. The concentrations of Mg(2+) and betaine were found to be important to the LAMP assay. The detection limits of both qualitative and quantitative LAMP for MON863 were as low as 4 haploid genomic DNA, and the LAMP reactions can be completed within 1 h at an isothermal temperature of 65 °C. CONCLUSION: The results of this study demonstrate that this new SYBR Green I-based quantitative LAMP assay system is reliable, sensitive and accurate.

A-T linker adapter polymerase chain reaction for determining flanking sequences by rescuing inverse PCR or thermal asymmetric interlaced PCR products.

The polymerase chain reaction (PCR)-based genome walking method has been extensively used to isolate unknown flanking sequences, whereas nonspecific products are always inevitable. To resolve these problems, we developed a new strategy to isolate the unknown flanking sequences by combining A-T linker adapter PCR with inverse PCR (I-PCR) or thermal asymmetric interlaced PCR (TAIL-PCR). The result showed that this method can be efficiently achieved with the flanking sequence from the Arabidopsis mutant and papain gene. Our study provides researchers with an additional method for determining genomic DNA flanking sequences to identify the target band from bulk of bands and to eliminate the cloning step for sequencing.

Effects of the adjunctive therapy of Sun's abdominal acupuncture on acute ischemic stroke and inflammatory factors: a randomized controlled trial. / å­™æ°è ¹é’ˆè¾ åŠ©æ²»ç–—æ€¥æ€§è„‘æ¢—æ­»åŠå¯¹ç‚Žæ€§å› å­çš„å½±å“ï¼šéšæœºå¯¹ç §è¯•éªŒ.

OBJECTIVES: To compare the clinical effect on acute ischemic stroke (AIS) between the combined treatment of Sun's abdominal acupuncture combined with the routine acupuncture and the simple routine acupuncture, and explore the influences on inflammatory factors i.e. interleukin (IL)-1ß and IL-10. METHODS: Eighty patients with AIS were randomly divided into an observation group (40 cases, 1 case dropped out) and a control group (40 cases, 1 case dropped out). The routine regimens of western medicine were administered in the two groups. In addition, the routine scalp acupuncture and the body acupuncture were used in the patients of the control group. The scalp acupuncture stimulation region and acupoints included the anterior parietal-temporal oblique line, Hegu (LI 4), Chize (LU 5), Shousanli (LI 10), etc. of affected side. In the observation group, on the base of the intervention of the control group, electroacupuncture was applied to "fouth abdominal area" of Sun's abdominal acupuncture, with the continuous wave and the frequency of 5 Hz. In the two groups, the intervention was given twice per day, once in every morning and afternoon separately, with the needles retained for 40 min in each intervention. The interventions were delivered for 6 days a week, lasting 3 weeks. The scores of Fugl-Meyer assessment scale (FMA), Berg balance scale (BBS) and the modified Barthel index (MBI), and the levels of IL-1ß and IL-10 in the serum were observed before and after treatment in the two groups; the effect and safety of interventions were compared between the two groups. RESULTS: After treatment, the scores of FMA, BBS and MBI increased in comparison with those before treatment in the two groups (P<0.01), and these scores in the observation group were higher than those in the control group (P<0.05, P<0.01). After treatment, in the two groups, the levels of IL-1ß in the serum were reduced in comparison with those before treatment (P<0.01), and the IL-1ß level in the observation group was lower than that in the control group (P<0.05); the levels of IL-10 in the serum were elevated in comparison with those before treatment in the two groups (P<0.01) and the IL-10 level in the observation group was higher than that in the control group (P<0.05). The total effective rate was 92.3% (36/39) in the observation group, which was superior to that in the control group (84.6% [33/39], P<0.05). There was no significant difference in the incidence of adverse reactions between the two groups (P>0.05). CONCLUSIONS: Sun's abdominal acupuncture combined with the routine acupuncture can ameliorate the motor impairment, adjust the balance dysfunction and improve the activities of daily living in the patients with AIS. The therapeutic effect of this combined regimen is better than that of the routine acupuncture, which may be associated with the regulation of the inflammatory factors after cerebral infarction.

Professor SUN Shentian's clinical experience in treatment of qiaobi with acupuncture. / 孙申田针灸治疗窍闭临床经验荟萃.

The paper introduces the clinical experience of professor SUN Shentian in treatment of qiaobi (orifice obstruction) with acupuncture. In pathogenesis, qiaobi refers to the obstruction of qi movement and abnormal ascending and descending of qi. "Obstruction of zangfu orifices" is the biao (symptoms), while "obstruction of brain orifice" is ben (root cause) of diseases. In the principle of treatment, bian and ben should be considered simultaneously, and it is proposed to regulating the spirit and unblocking the orifices in treatment. Baihui (GV 20) and the cognitive emotional area on the head are selected for regulating the spirit and opening the brain orifice, focusing on the root cause of diseases. Based on the connection between the symptoms and the cerebral cortical functions, the different points on the head are used, the Siguan points (Hegu [LI 4] and Taichong [LR 3]) and yuan-source points combined, and the acupoint prescription is composed according to the interior-exterior relationship of meridians. With repetitive transcranial acupuncture stimulation at the points on the head, the therapeutic effect is strengthened. It provides a novel approach to treatment of diseases related to qiaobi.

[Repetitive transcranial acupuncture stimulation combined with electroacupuncture in treatment of acute facial palsy with retroauricular pain: a randomized controlled trial].

OBJECTIVE: To observe the clinical effect of repetitive transcranial acupuncture stimulation (rTAS) combined with electroacupuncture (EA) in treatment of acute facial palsy with retroauricular pain. METHODS: Sixty-eight patients of acute facial palsy with retroauricular pain were randomly divided into an observation group (34 cases, 3 cases dropped out) and a control group (34 cases, 3 cases dropped out). On the basis of conventional therapy, in the control group, Yangbai (GB 14), Cuanzhu (BL 2), Sibai (ST 2), Quanliao (SI 18), Dicang (ST 4), Yifeng (TE 17), Qianzheng (Extra point) and Taiyang (EX-HN 5) on the affected side, and bilateral Hegu (LI 4) were selected. EA was attached to Yangbai (GB 14) and Cuanzhu (BL 2), and Sibai (ST 2) and Dicang (ST 4), respectively, using intermittent wave. In the observation group, on the basis of the regimen as the control group, rTAS was delivered at Baihui (GV 20) and the 1/5 of the lower motor area on the bilateral sides; EA of dense wave was given at the sites of the mastoidⅠand Ⅱ. The intervention of each group was delivered once a day, 6 times a week as one course for 4 courses and taking a day off every course. Before treatment and at the moment after the first treatment completion, the score of visual analogue scale (VAS) was observed in the two groups and the days of retroauricular pain were recorded. Before and after treatment, the score of Sunnybrook facial grading system (SFGS), the grade of House-Brackmann facial nerve function evaluation system (H-B), the latency and amplitude of the motor conduction from the foramina stylomastoideum to the frontal muscle, the orbicularis oris muscle and the orbicularis oculi muscle on the affected facial nerve, were observed in the patients of two groups and the clinical effect was compared between the two groups after treatment. RESULTS: After treatment, SFGS score was increased (P<0.05), H-B grade was improved (P<0.05), the latency was shortened in the motor conduction from the foramina stylomastoideum to the frontal muscle, the orbicularis oris muscle and the orbicularis oculi muscle on the affected facial nerve (P<0.05) and its amplitude elevated (P<0.05) when compared with those before treatment in the two groups. In the observation group, SFGS score was higher (P<0.05), H-B grade was superior (P<0.05), the latency was shorter in the motor conduction from the foramina stylomastoideum to the frontal muscle, the orbicularis oris muscle and the orbicularis oculi muscle on the affected facial nerve (P<0.05) and its amplitude was higher (P<0.05) when compared with those of the control group after treatment. After the completion of the first treatment, VAS score of either group was reduced in comparison with that before treatment (P<0.05), and the score in the observation group was lower than that of the control group (P<0.05). The duration of retroauricular pain was shortened in the observation group when compared with that of the control group (P<0.05). The total effective rate was 87.1% (27/31) in the observation group, which was higher than 77.4% (24/31) of the control group (P<0.05). CONCLUSION: The rTAS combined with EA is effective for reducing neurologic impairment of acute facial palsy and alleviating retroauricular pain in the patients.

Elucidating the interaction mechanism of rice glutelin and soybean 11S globulin using multi-spectroscopy and molecular dynamics simulation methods.

Rice gluten, as the hydrophobic protein, exhibits restricted application value in hydrophilic food, which may be enhanced through interaction with soybean 11S globulin, characterized by favorable functional properties. This study aims at revealing their interaction mechanism via multi-spectroscopy and molecular dynamics simulation. The formation and structural change of rice glutelin-soybean 11S globulin complexes were detected using fluorescence, ultra-violet and circular dichroism spectra. The addition of 11S globulin increased the contents of α-helix, ß-turn and random coil, but decreased ß-sheet content, and the change in secondary structure was correlated with particle size. Moreover, exposure of hydrophobic groups and formation of disulfide bonds occurred in the complexes. Molecular dynamics simulation verified these experimental results through analyses of root mean square deviation and fluctuation, hydrogen bond, secondary structure, and binding free energy analysis. This study contributes to expounding the interaction mechanism of protein and protein from the molecular level.

Single universal primer multiplex ligation-dependent probe amplification with sequencing gel electrophoresis analysis.

In this study, a novel single universal primer multiplex ligation-dependent probe amplification (SUP-MLPA) technique that uses only one universal primer to perform multiplex polymerase chain reaction (PCR) was developed. Two reversely complementary common sequences were designed on the 5' or 3' end of the ligation probes (LPs), which allowed the ligation products to be amplified through only a single universal primer (SUP). SUP-MLPA products were analyzed on sequencing gel electrophoresis with extraordinary resolution. This method avoided the high expenses associated with capillary electrophoresis, which was the commonly used detection instrument. In comparison with conventional multiplex PCR, which suffers from low sensitivity, nonspecificity, and amplification disparity, SUP-MLPA had higher specificity and sensitivity and a low detection limit of 0.1 ng for detecting single crop species when screening the presence of genetically modified crops. We also studied the effect of different lengths of stuffer sequences on the probes for the first time. Through comparing the results of quantitative PCR, the LPs with different stuffer sequences did not affect the ligation efficiency, which further increased the multiplicity of this assay. The improved SUP-MLPA and sequencing gel electrophoresis method will be useful for food and animal feed identification, bacterial detection, and verification of genetic modification status of crops.

[Repeated transcranial acupuncture combined with electroacupuncture for flat descending sudden deafness: a randomized controlled trial].

OBJECTIVE: To observe the clinical effect of repeated transcranial acupuncture combined with electroacupuncture on flat descending sudden deafness. METHODS: A total of 80 patients with flat descending sudden deafness were randomly divided into an acupuncture and medication group (39 cases, 1 case dropped off) and a western medication group (39 cases, 1 case dropped off). Patients in the western medication group were treated with glucocorticoid + batroxobin + ginkgo leaf preparation. On the basis of the western medication group, patients in the acupuncture and medication group were treated with acupuncture at Baihui (GV 20), Ningshen, Yunting area, Mastoid 1 point, Mastoid 2 point, Fengchi (GB 20), Gongxue, Tinghui (GB 2), etc., repeated transcranial acupuncture was applied at Baihui (GV 20), Ningshen and Yunting area; Mastoid 1 point and Mastoid 2 point, Fengchi (GB 20) and Gongxue of the affected side were connected to the electroacupuncture instrument, continuous wave, 2 Hz in frequency. The needle was retained for 30 min, once a day, and rest for 1 d after 6 d of continuous treatment. All patients were treated for 3 weeks. The average hearing threshold, tinnitus handicap inventory (THI) score, dizziness handicap inventory (DHI) score and speech discrimination test score were compared between the two groups before and after treatment, and the clinical effect was evaluated. RESULTS: After treatment, the average hearing threshold, THI and DHI scores of the two groups were lower than those before treatment (P<0.05), and above indexes in the acupuncture and medication group were lower than those in the western medication group (P<0.05). The speech discrimination test scores of the two groups were higher than those before treatment (P<0.05), and the score in the acupuncture and medication group was higher than that in the western medication group (P<0.05). The total effective rate was 87.2% (34/39) in the acupuncture and medication group, which was higher than 74.4% (29/39) in the western medication group (P<0.05). CONCLUSION: Repeated transcranial acupuncture combined with electroacupuncture can improve the hearing level of patients with flat descending sudden deafness, relieve tinnitus and vertigo symptoms.

Effect of cold plasma on breaking activated sludge and the output dominance of protein.

Upon contacting with water, cold plasma should produce numerous ozone molecules and free electrons at room temperature. In this study, a cold plasma generator was used to break the walls of residual activated sludge obtained from domestic sewage. The impact was mainly influenced by the ozone generated. With 800 W power, sludge wastewater pH of 12.0, and under continuous treatment for 10 h, the system's reduction efficiency for the dry sludge was ≈90%. Furthermore, the organic matter content (especially protein) of the upper layer of the sludge solution increased a lot after the sludge digestion. This observation proved the reduction of sludge from both sides. Moreover, when the cold plasma technique was compared with thermal acid hydrolysis, thermal alkali hydrolysis, and ultrasonication for extracting protein from activated sludge, cold plasma wall-breaking sludge exhibited the highest efficiency, reaching 38.2% under ambient temperature. After the analysis, the toxic metal content in the extracted protein was near zero, which is a level other protein extraction methods via sludge breaking have not achieved to date, we attribute this efficiency to free electrons the cold plasma produce. These species promote the transformation of metal ions into atomic metals, thereby facilitating their removal.

[SUN Shen-tian's clinical experience in treating Tourette's syndrome with acupuncture].

To introduce the clinical experience of professor SUN Shen-tian in treatment of Tourette's syndrome (TS) with acupuncture. TS is a psychosomatic disease and the core pathogenesis refers to blood deficiency producing internal wind. The disease is located in the heart and liver. Acupoints are selected according to the functional orientation of the cerebral cortex. The extrapyramidal system area is preferred for tic disorder, and the emotional area is for behavioral disorder. The treatment focuses on regulating the mind by multiple methods, including manual needling technique used the transcranial repeated acupuncture, and regulating the mind by taking multiple acupoints, Baihui (GV 20), Neiguan (PC 6), Shenmen (HT 7) and Dazhong (KI 4) are stimulated. For abdominal twitching and mental symptoms of TS children, the first and third abdominal areas are selected. The target symptoms (biao) are treated specially by local acupoints, the combination of the starting and ending acupoints of the affected meridian, or the acupoints of the meridians with same name. The modified chaihu longgu muli decoction and siwu decoction are prescribed to sooth liver, nourish blood and soothe wind. In association with the characteristics and target symptoms of TS, the sequential therapy is used with filiform needling, intradermal needling, Chinese herbal medication and psychotherapy.

[GAO Wei-bin's clinical experience in treatment of neurogenic bladder with acupuncture].

The paper introduces GAO Wei-bin's clinical experience in acupuncture treatment for neurogenic bladder. In association with the etiology, the location and types of neurogenic bladder and in accordance with nerve anatomy and meridian differentiation, the acupoints are selected accurately in treatment. Four acupoint prescriptions are allocated. For frequent urination and urinary incontinence, the foot-motor-sensory area of scalp acupuncture, Shenshu (BL 23) and Huiyang (BL 35) are used. For all kinds of urine retention, especially the patients who are not suitable for acupuncture at the lumbar region, Zhongji (CV 3), Qugu (CV 2), Henggu (KI 11) and Dahe (KI 12) are selected. For all kinds of urine retention, Zhongliao (BL 33) and Ciliao (BL 32) are applicable. For the patients with both dysuria and urinary incontinence, Zhongliao (BL 33), Ciliao (BL 32) and Huiyang (BL 35) are chosen. In treatment of neurogenic bladder, both biao (root causes) and ben (primary symptoms) are considered, as well as the accompanying symptoms; and electroacupuncture is combined accordingly. During the delivery of acupuncture, the sites where the acupoints located are detected and palpated so as to rationally control the depth of needle insertion and the operation of reinforcing and reducing needling techniques.

The impact of mass gatherings on the local transmission of COVID-19 and the implications for social distancing policies: Evidence from Hong Kong.

Mass gatherings provide conditions for the transmission of infectious diseases and pose complex challenges to public health. Faced with the COVID-19 pandemic, governments and health experts called for suspension of gatherings in order to reduce social contact via which virus is transmitted. However, few studies have investigated the contribution of mass gatherings to COVID-19 transmission in local communities. In Hong Kong, the coincidence of the relaxation of group gathering restrictions with demonstrations against the National Security Law in mid-2020 raised concerns about the safety of mass gatherings under the pandemic. Therefore, this study examines the impacts of mass gatherings on the local transmission of COVID-19 and evaluates the importance of social distancing policies. With an aggregated dataset of epidemiological, city-level meteorological and socioeconomic data, a Synthetic Control Method (SCM) is used for constructing a 'synthetic Hong Kong' from over 200 Chinese cities. This counterfactual control unit is used to simulate COVID-19 infection patterns (i.e., the number of total cases and daily new cases) in the absence of mass gatherings. Comparing the hypothetical trends and the actual ones, our results indicate that the infection rate observed in Hong Kong is substantially higher than that in the counterfactual control unit (2.63% vs. 0.07%). As estimated, mass gatherings increased the number of new infections by 62 cases (or 87.58% of total new cases) over the 10-day period and by 737 cases (or 97.23%) over the 30-day period. These findings suggest the necessity of tightening social distancing policies, especially the prohibition on group gathering regulation (POGGR), to prevent and control COVID-19 outbreaks.