IMPATIENT-qPCR: monitoring SELEX success during in vitro aptamer evolution.

SELEX (Systematic Evolution of Ligands by Exponential enrichment) processes aim on the evolution of high-affinity aptamers as binding entities in diagnostics and biosensing. Aptamers can represent game-changers as constituents of diagnostic assays for the management of instantly occurring infectious diseases or other health threats. Without in-process quality control measures SELEX suffers from low overall success rates. We present a quantitative PCR method for fast and easy quantification of aptamers bound to their targets. Simultaneous determination of melting temperatures (Tm) of each SELEX round delivers information on the evolutionary success via the correlation of increasing GC content and Tm alone with a round-wise increase of aptamer affinity to the respective target. Based on nine successful and published previous SELEX processes, in which the evolution/selection of aptamer affinity/specificity was demonstrated, we here show the functionality of the IMPATIENT-qPCR for polyclonal aptamer libraries and resulting individual aptamers. Based on the ease of this new evolution quality control, we hope to introduce it as a valuable tool to accelerate SELEX processes in general. IMPATIENT-qPCR SELEX success monitoring. Selection and evolution of high-affinity aptamers using SELEX technology with direct aptamer evolution monitoring using melting curve shifting analyses to higher Tm by quantitative PCR with fluorescence dye SYBR Green I. KEY POINTS: • Fast and easy analysis. • Universal applicability shown for a series of real successful projects.

Robust Fluorometric Aptamer Assay for Direct and Rapid Detection of Clinical Isolates of Candida spec.

Infections caused by yeasts of the genus Candida are likely to occur not only in immunocompromised patients but also in healthy individuals, leading to infections of the gastrointestinal tract, urinary tract, and respiratory tract. Due to the rapid increase in the frequency of reported Candidiasis cases in recent years, diagnostic research has become the subject of many studies, and therefore, we developed a polyclonal aptamer library-based fluorometric assay with high specificity and affinity towards Candida spec. to quantify the pathogens in clinical samples with high sensitivity. We recently obtained the specific aptamer library R10, which explicitly recognized Candida and evolved it by mimicking an early skin infection model caused by Candida using the FluCell-SELEX system. In the follow-up study presented here, we demonstrate that the aptamer library R10-based bioassay specifically recognizes invasive clinical Candida isolates, including not only C. albicans but also strains like C. tropcialis, C. krusei, or C. glabrata. The next-generation fluorometric bioassay presented here can reliably and easily detect an early Candida infection and could be used for further clinical research or could even be developed into a full in vitro diagnostic tool.

A Polyclonal Aptamer Library for the Specific Binding of the Gut Bacterium Roseburia intestinalis in Mixtures with Other Gut Microbiome Bacteria and Human Stool Samples.

Roseburia intestinalis has received attention as a potential probiotic bacterium. Recent studies have demonstrated that changes in its intestinal abundance can cause various diseases, such as obesity, enteritis and atherosclerosis. Probiotic administration or fecal transplantation alter the structure of the intestinal flora, offering possibilities for the prevention and treatment of these diseases. However, current monitoring methods, such as 16S rRNA sequencing, are complex and costly and require specialized personnel to perform the tests, making it difficult to continuously monitor patients during treatment. Hence, the rapid and cost-effective quantification of intestinal bacteria has become an urgent problem to be solved. Aptamers are of emerging interest because their stability, low immunogenicity and ease of modification are attractive properties for a variety of applications. We report a FluCell-SELEX polyclonal aptamer library specific for R. intestinalis isolated after seven evolution rounds, that can bind and label this organism for fluorescence microscopy and binding assays. Moreover, R. intestinalis can be distinguished from other major intestinal bacteria in complex defined mixtures and in human stool samples. We believe that this preliminary evidence opens new avenues towards aptamer-based electronic biosensors as new powerful and inexpensive diagnostic tools for the relative quantitative monitoring of R. intestinalis in gut microbiomes.

Identification of the Xyloglucan Endotransglycosylase/Hydrolase (XTH) Gene Family Members Expressed in Boehmeria nivea in Response to Cadmium Stress.

Xyloglucan endotransglycosylase/hydrolase (XTH) genes play an important role in plant resistance to abiotic stress. However, systematic studies of the response of Boehmeria nivea (ramie) XTH genes (BnXTHs) to cadmium (Cd) stress are lacking. We sought to identify the BnXTH-family genes in ramie through bioinformatics analyses and to investigate their responses to Cd stress. We identified 19 members of the BnXTH gene family from the ramie genome, referred to as BnXTH1-19, among which BnXTH18 and BnXTH19 were located on no chromosomes and the remaining genes were unevenly distributed across 11 chromosomes. The 19 members were divided into four groups, Groups I/II/IIIA/IIIB, according to their phylogenetic relationships, and these groups were supported by analyses of intron-exon structure and conserved motif composition. A highly conserved catalytic site (HDEIDFEFLG) was observed in all BnXTH proteins. Additionally, three gene pairs (BnXTH6-BnXTH16, BnXTH8-BnXTH9, and BnXTH17-BnXTH18) were obtained with a fragment and tandem-repeat event analysis of the ramie genome. An analysis of cisregulatory elements revealed that BnXTH expression might be regulated by multiple hormones and abiotic and biotic stress responses. In particular, 17 cisregulatory elements related to abiotic and biotic stress responses and 11 cisregulatory elements related to hormone responses were identified. We also found that most BnXTH genes responded to Cd stress, and BnXTH1, BnXTH3, BnXTH6, and BnXTH15 were most likely to contribute to the Cd tolerance of ramie, as evidenced by the substantial increases in expression under Cd treatment. Heterologous expression of BnXTH1, BnXTH6, and BnXTH15 significantly enhanced the Cd tolerance of transgenic yeast cells. These results suggest that the BnXTH gene family is involved in Cd stress responses, laying a theoretical foundation for functional studies of BnXTH genes and the innovative breeding of Cd-tolerant ramie.

Polyclonal Aptamer Libraries from a FluRoot-SELEX for the Specific Labeling of the Apical and Elongation/Differentiation Zones of Arabidopsis thaliana Roots.

In more than 30 years of aptamer research, it has become widely accepted that aptamers are fascinating binding molecules for a vast variety of applications. However, the majority of targets have been proteins, although special variants of the so-called SELEX process for the molecular evolution of specific aptamers have also been developed, allowing for the targeting of small molecules as well as larger structures such as cells and even cellular networks of human (tumor) tissues. Although the provocative thesis is widely accepted in the field, that is, in principle, any level of complexity for SELEX targets is possible, the number of studies on whole organs or at least parts of them is limited. To pioneer this thesis, and based on our FluCell-SELEX process, here, we have developed polyclonal aptamer libraries against apices and the elongation/differentiation zones of plant roots as examples of organs. We show that dedicated libraries can specifically label the respective parts of the root, allowing us to distinguish them in fluorescence microscopy. We consider this achievement to be an initial but important evidence for the robustness of this SELEX variant. These libraries may be valuable tools for plant research and a promising starting point for the isolation of more specific individual aptamers directed against root-specific epitopes.

A Polyclonal Selex Aptamer Library Directly Allows Specific Labelling of the Human Gut Bacterium Blautia producta without Isolating Individual Aptamers.

Recent studies have demonstrated that changes in the abundance of the intestinal bacterium Blautia producta, a potential probiotic, are closely associated with the development of various diseases such as obesity, diabetes, some neurodegenerative diseases, and certain cancers. However, there is still a lack of an effective method to detect the abundance of B. producta in the gut rapidly. Especially, DNA aptamers are now widely used as biometric components for medical testing due to their unique characteristics, including high chemical stability, low production cost, ease of chemical modification, low immunogenicity, and fast reproducibility. We successfully obtained a high-affinity nucleic acid aptamer library (B.p-R14) after 14 SELEX rounds, which efficiently discriminates B. producta in different analysis techniques including fluorometric suspension assays or fluorescence microscopy from other major gut bacteria in complex mixtures and even in human stool samples. These preliminary findings will be the basis towards aptamer-based biosensing applications for the fast and reliable monitoring of B. producta in the human gut microbiome.

Transcallosal-lateral ventricle-choroid fissure approach for excising large pineal region tumors extending into the third ventricle: experience in 15 pediatric cases.

Pineal region is deep located and tightly connected with surrounding important nerves, blood vessels, and other critical structure. Tumors in the regions are more commonly observed in children with complex pathology, difficult surgery, and poor prognosis. However, excision surgery on large tumor in the pineal region and extending into the third ventricle is characterized by high difficulty and uneasy treatment. Related experience and knowledge about excising large tumor in children pineal region which extending into the third ventricle by transcallosal-lateral ventricle-choroid fissure approach was described in the paper. Clinical data of 15 children patients with large tumors in pineal region which extending into the third ventricle treated by our group from 2011 to 2015 was retrospectively analyzed (< 15 years of age, tumor size > >3 × 3 × 3 cm3). Fifteen cases underwent tumor excision with surgery by transcallosal-lateral ventricle-choroid fissure approach including 11 male patients and 4 female patients with average age of 9.1 years and tumor volume from 3 × 3 × 3 to 5 × 6 × 7 cm3. They were assessed according to postoperative clinical symptom remission rate, blood loss during surgery, tumor removal rate, postoperative recurrence rate, etc. as indicators. Five cases belonged to germ cell tumors, 4 for teratoma, 1 for malignant teratoma, 2 for glioma, 2 for meningioma, and 1 for pineocytoma. Surgical total excision rate was 100%, while the mean intraoperative blood loss was about 300 ml. No case was dead due to surgery, and all cases had good postoperative recovery. Children patients are characterized by less blood than adults, larger volume size during surgery treatment, poor postoperative anti-infectious ability, etc. High requirements are proposed on surgery treatment, by long-term clinical exploration and experience accumulation. Our group believes that surrounding structures such as veins and dome are stretched and stimulated rarely in excision of children in large and medium tumor in pineal region which extending into the third ventricle by transcallosal-lateral ventricle-choroid fissure approach; thereby, the tumor can be completely excised, and larger veins and other important structures can be well-preserved with fewer complications, high full cut rate, and rapid recovery.

BSA Hydrogel Beads Functionalized with a Specific Aptamer Library for Capturing Pseudomonas aeruginosa in Serum and Blood.

Systemic blood stream infections are a major threat to human health and are dramatically increasing worldwide. Pseudomonas aeruginosa is a WHO-alerted multi-resistant pathogen of extreme importance as a cause of sepsis. Septicemia patients have significantly increased survival chances if sepsis is diagnosed in the early stages. Affinity materials can not only represent attractive tools for specific diagnostics of pathogens in the blood but can prospectively also serve as the technical foundation of therapeutic filtration devices. Based on the recently developed aptamers directed against P. aeruginosa, we here present aptamer-functionalized beads for specific binding of this pathogen in blood samples. These aptamer capture beads (ACBs) are manufactured by crosslinking bovine serum albumin (BSA) in an emulsion and subsequent functionalization with the amino-modified aptamers on the bead surface using the thiol- and amino-reactive bispecific crosslinker PEG4-SPDP. Specific and quantitative binding of P. aeruginosa as the dedicated target of the ACBs was demonstrated in serum and blood. These initial but promising results may open new routes for the development of ACBs as a platform technology for fast and reliable diagnosis of bloodstream infections and, in the long term, blood filtration techniques in the fight against sepsis.

FluCell-SELEX Aptamers as Specific Binding Molecules for Diagnostics of the Health Relevant Gut Bacterium Akkermansia muciniphila.

Based on their unique properties, oligonucleotide aptamers have been named a gift of biological chemistry to life science. We report the development of DNA aptamers as the first high-affinity binding molecules available for fast and rapid labeling of the human gut bacterium Akkermansia muciniphila with a certain impact on Alzheimer´s disease. Fast and reliable analyses of the composition of microbiomes is an emerging field in microbiology. We describe the molecular evolution and biochemical characterization of a specific aptamer library by a FluCell-SELEX and the characterization of specific molecules from the library by bioinformatics. The aptamer AKK13.1 exerted universal applicability in different analysis techniques in modern microbiology, including fluorimetry, confocal laser scanning microscopy and flow cytometry. It was also functional as a specific binding entity hybridized to anchor primers chemically coupled via acrydite-modification to the surface of a polyacrylamide-hydrogel, which can be prototypically used for the construction of affinity surfaces in sensor chips. Together, the performance and methodological flexibility of the aptamers presented here may open new routes not only to develop novel Akkermansia-specific assays for clinical microbiology and the analyses of human stool samples but may also be an excellent starting point for the construction of novel electronic biosensors.

[Long Non-coding RNA Act as Regulators of Autophagy in Disease Treatment].

Long non-coding RNA (lncRNA) refer to transcripts longer than 200 nucleotides that have a low coding potential. Autophagy,a unique life phenomenon of eukaryotic cells,removes excess or damaged organelles during cell growth and development and plays a key role in maintaining homeostasis. As a key regulator of cellular metabolism,lncRNA are involved in disease treatment by regulating autophagy. This article summarizes the role of lncRNA in the treatment of cancer,cardiovascular and cerebrovascular diseases,neurodegenerative diseases,and bacterial infections and analyzes the molecular mechanisms of lncRNA in regulating autophagy,along with prospects of its applications in other areas.

[Multi Spectral Detection of Ethanol Content in Gasoline Based on SiPLS Feature Extraction and Information Fusion].

The ethanol content in ethanol gasoline was detected with ultraviolet/visible(UV/vis) and near-infrared (NIR) spectroscopy while information fusion technology and synergy interval PLS(SiPLS) algorithm were used as the feature extraction method with the establishment of partial least squares(PLS) regression model. Using the information fusion theory, UV/vis and NIR spectra were used for data fusion, the data level fusion (Low level data fusion, LLDF) and feature level fusion(Mid-level data fusion, MLDF) model were established. The results were compared with the single source modelwith low level data fusion before vector normalization(LLDF-VN1) selected for the optimal model. Finally, the optimal model was tested using the spectral data collected from the samples of high ethanol content and commercial gasoline. The results showed that both UV/vis and NIR can be used to detect and provide good prediction results, whereas direct fusion of the UV/vis and NIR spectral data provided the best results in the regression model based on the calibration set, with the highest correlation coefficient rc, the smallest Biasc and RMSECV values, as 0.999 9, 0.125 8 and 0.000 6, respectively. And the prediction effect of the model of LLDF-VN1(low level data fusion before vector normalization) was the best, r(p)=0.999 1，Bias(p)=0.352 7，RMSEP=-0.073 8. In the verification of the optimal model (LLDF-VN1) by the self distribution solution, rp=0.999 7, Bias(p)=0.102 2, RMSEP=0.329 1; and that for gasoline sold on market, r(p)=0.990 1, RMSEP=0.675 1, Bias(p)=0.892 7, respectively. It showed that the data level fusion based on UV/vis and NIR spectral information could be used to detect the content of ethanol in ethanol-gasoline quickly and accurately, achieving a wide range of ethanol concentration detection, which laid a foundation for further realization of the rapid detection of substances in the blended fuel oil.

[An Extraction and Recognition Method of the Distributed Optical Fiber Vibration Signal Based on EMD-AWPP and HOSA-SVM Algorithm].

Given that the traditional signal processing methods can not effectively distinguish the different vibration intrusion signal, a feature extraction and recognition method of the vibration information is proposed based on EMD-AWPP and HOSA-SVM, using for high precision signal recognition of distributed fiber optic intrusion detection system. When dealing with different types of vibration, the method firstly utilizes the adaptive wavelet processing algorithm based on empirical mode decomposition effect to reduce the abnormal value influence of sensing signal and improve the accuracy of signal feature extraction. Not only the low frequency part of the signal is decomposed, but also the high frequency part the details of the signal disposed better by time-frequency localization process. Secondly, it uses the bispectrum and bicoherence spectrum to accurately extract the feature vector which contains different types of intrusion vibration. Finally, based on the BPNN reference model, the recognition parameters of SVM after the implementation of the particle swarm optimization can distinguish signals of different intrusion vibration, which endows the identification model stronger adaptive and self-learning ability. It overcomes the shortcomings, such as easy to fall into local optimum. The simulation experiment results showed that this new method can effectively extract the feature vector of sensing information, eliminate the influence of random noise and reduce the effects of outliers for different types of invasion source. The predicted category identifies with the output category and the accurate rate of vibration identification can reach above 95%. So it is better than BPNN recognition algorithm and improves the accuracy of the information analysis effectively.

[A Multi-Peak Brillouin Scattering Spectrum Feature Extraction Method Based on Multi-Criteria Decision-Making and Particle Swarm Optimization-Levenberg Marquardt Hybrid Optimization Algorithm].

As to fitting the multi-peaks Brillouin scattering spectrum with traditional method, the maximum power point is usually selected as the benchmark while other extreme value points which are less than the maximum power are lost. The fitting curve only has one peak because the multi-peaks Brillouin scattering spectrum is simplified into the highest peak and several small peaks. So it will lead to the loss of useful information. In order to improve the feature extraction accuracy of Brillouin scattering spectrum, a hybrid optimization algorithm named MCDM-PSO-LM algorithm is presented based on MCDM and PSO-LM algorithm. The MCDM algorithm can identify and locate the peaks and valleys of multi-peaks Brillouin scattering spectrum accurately. The PSO-LM hybrid algorithm can realize the curve fitting on every peak and valley, and it can seach the center frequency shift of each peak. The PSO-LM hybrid algorithm can solves these disadvantages, which PSO algorithm premature convergence to local minimum and LM algorithm depends on the initial value problem. It can also combine the global search ability of PSO algorithm and the local search ability of LM algorithm. Compared with traditional algorithms, MCDM-PSO-LM algorithm can ensure the solving speed and accuracy to the optimal value, and the analytical solution will be close to the optimal value sufficiently. So it improves the operation ability. With different signal to noise ratio and linewidth, the results of frequency shift and temperature error show that the MCDM-PSO-LM method can locate every peak and valley of multi-peaks Brillouin scattering spectrum accurately. Thus, it can be used for the feature extraction of multi-peaks Brillouin scattering spectrum. The recognition effect of this method is obviously better than that of traditional algorithms and it can improve the accuracy of information analysis.

[Refractive Index Insensitive Temperature Sensor Based on Cascading Single Mode Fiber with Few Mode Fiber].

A refractive index insensitive temperature sensor is proposed base on cascading single mode fiber with few mode fiber(FMF). During the sensor preparation, the splicing current is set to 100 mA, and a section of FMF is no core-offset splicing between two single-mode fibers. Therefore, it can motivate the transmission mode preferably and form optical fiber Mach-Zehnder interferometer. The mode phase difference in FMF will be changed according to the outside environment. It will cause interference fringe shift. The parameter to be measured can be achieved by detecting the amount shift of interference spectrum. The FMF can transmit four modes with LP01, LP11, LP21, LP02. The transmission spectrum is also analyzed, which shows that they have two modes of LP01 and LP11 in sensor with the length of 81.5 mm. In the refractive index and temperature sensing experiment, the cascading FMF sensor with the length of 81.5 mm is used. The results show that the transmission spectrum of sensor appears obvious blue shift as temperature is increasing, the temperature sensitivity can be up to -85.9 pm·â„ƒ-1 within the range of 27.6~93.8 ℃ with good linearity. The refractive index sensitivity is 3.697 34 nm·RIU-1 within the range of 1.347 1~1.443 9. There is no obvious shift phenomenon in the transmission spectrum with the feature of refractive index insensitive. Therefore, compared with the traditional cladding mode and multimode interferometric fiber-optic sensor, the proposed sensor based on FMF is easier to control and analyze transmission mode has the advantages of simple structure, easy process and high sensitivity. It can avoid cross-sensitivity between temperature and refractive index measurement. Thus, it can be used for temperature detection of power system, biomedicine, aerospace and other fields.

[The High Precision Analysis Research of Multichannel BOTDR Scattering Spectral Information Based on the TTDF and CNS Algorithm].

Traditional BOTDR optical fiber sensing system uses single channel sensing fiber to measure the information features. Uncontrolled factors such as cross-sensitivity can lead to a lower scattering spectrum fitting precision and make the information analysis deflection get worse. Therefore, a BOTDR system for detecting the multichannel sensor information at the same time is proposed. Also it provides a scattering spectrum analysis method for multichannel Brillouin optical time-domain reflection (BOT-DR) sensing system in order to extract high precision spectrum feature. This method combines the three times data fusion (TTDF) and the cuckoo Newton search (CNS) algorithm. First, according to the rule of Dixon and Grubbs criteria, the method uses the ability of TTDF algorithm in data fusion to eliminate the influence of abnormal value and reduce the error signal. Second, it uses the Cuckoo Newton search algorithm to improve the spectrum fitting and enhance the accuracy of Brillouin scattering spectrum information analysis. We can obtain the global optimal solution by smart cuckoo search. By using the optimal solution as the initial value of Newton algorithm for local optimization, it can ensure the spectrum fitting precision. The information extraction at different linewidths is analyzed in temperature information scattering spectrum under the condition of linear weight ratio of 1:9. The variances of the multichannel data fusion is about 0.0030, the center frequency of scattering spectrum is 11.213 GHz and the temperature error is less than 0.15 K. Theoretical analysis and simulation results show that the algorithm can be used in multichannel distributed optical fiber sensing system based on Brillouin optical time domain reflection. It can improve the accuracy of multichannel sensing signals and the precision of Brillouin scattering spectrum analysis effectively.

[A Brillouin Scattering Spectrum Feature Extraction Based on Flies Optimization Algorithm with Adaptive Mutation and Generalized Regression Neural Network].

According to the high precision extracting characteristics of scattering spectrum in Brillouin optical time domain reflection optical fiber sensing system, this paper proposes a new algorithm based on flies optimization algorithm with adaptive mutation and generalized regression neural network. The method takes advantages of the generalized regression neural network which has the ability of the approximation ability, learning speed and generalization of the model. Moreover, by using the strong search ability of flies optimization algorithm with adaptive mutation, it can enhance the learning ability of the neural network. Thus the fitting degree of Brillouin scattering spectrum and the extraction accuracy of frequency shift is improved. Model of actual Brillouin spectrum are constructed by Gaussian white noise on theoretical spectrum, whose center frequency is 11.213 GHz and the linewidths are 40-50, 30-60 and 20-70 MHz, respectively. Comparing the algorithm with the Levenberg-Marquardt fitting method based on finite element analysis, hybrid algorithm particle swarm optimization, Levenberg-Marquardt and the least square method, the maximum frequency shift error of the new algorithm is 0.4 MHz, the fitting degree is 0.991 2 and the root mean square error is 0.024 1. The simulation results show that the proposed algorithm has good fitting degree and minimum absolute error. Therefore, the algorithm can be used on distributed optical fiber sensing system based on Brillouin optical time domain reflection, which can improve the fitting of Brillouin scattering spectrum and the precision of frequency shift extraction effectively.

Cross-reactivity between human cytomegalovirus peptide 981-1003 and myelin oligodendroglia glycoprotein peptide 35-55 in experimental autoimmune encephalomyelitis in Lewis rats.

Multiple sclerosis (MS) has been documented to have various clinical and pathological presentations. However the underlying mechanisms remain unknown. Viral infections may play a certain role in the etiopathogenesis of MS. This study was designed to explore whether different phospholipid peptides and viral mimic peptides induce antigen specific lesion in experimental autoimmune encephalomyelitis (EAE), an MS animal model. In the present study, Lewis rats immunized with myelin basic protein (MBP) 82-99 or MBP68-86 exhibited clinical signs of EAE and inflammatory infiltrates throughout CNS. Immunization with myelin oligodendroglia glycoprotein (MOG) 35-55 also induced inflammatory infiltrates in spinal cords. Although cytomegalovirus (CMV) 981-1003 failed to induce clinical signs of EAE and inflammatory infiltrates, immunological examination revealed that CMV981-1003 cross-reacted with serum from rats immunized with MOG35-55, and vice versa. Further, MOG35-55 triggered CMV981-1003 specific lymphocytes recruitment in spleen. Together these, this study provides certain evidences for various pathological manifestations of EAE and the linkage of viral mimic peptides with phospholipid peptides. Molecular mimicry may be an explanation the pathogenesis of MS.

Somatosensory reflex seizures in a patient with hypertrophic cranial pachymeningitis.

Somatosensory-evoked reflex epilepsy is characterized by seizures in response to specific stimuli. It is highly uncommon for somatosensory-evoked focal seizures to be caused by movement or a change in posture. Reflex epilepsy induced by both somatosensory and proprioceptive stimulations has not been previously reported. In this study, we present a case of reflex epilepsy evoked by somatosensory and proprioceptive stimulation in a patient with hypertrophic cranial pachymeningitis. After comparing our patient with other cases of previously reported somatosensory-evoked reflex epilepsy, we determined that our patient had an unusual cause of reflex epilepsy.

[The nitrate and temperature impact analysis in the detection of COD by UV spectrum].

Configured standard solution of chemical oxygen demand with potassium hydrogen phthalate was used as experimental subjects, collected ultraviolet absorption spectra of the standard solution in the range of 1,800 mg x L(-1), were collected, and PLS (partial least squares) algorithm was used to establish the correction model of different spectral region, the results showed that. The model in the spectral region of 265-310 nm had the highest correlation and smallest error; In order to eliminate the impact of nitrates and temperature on the detection of the COD , studied the changes of the UV absorption spectrum with different concentrations of sodium standard solution and different temperature. The results showed that absorption of nitrate in 208-238 nm was apparent, and the model for spectral region of 265-310 nm was free from the influence of nitrate; In the full range of spectrum, temperature rising leads to an increase in absorbance, thus the temperature compensation model was established for the different spectral region through predictive analysis.

Sustainable tourism development for traditional Chinese drama's intangible cultural heritage.

This study establishes an expert-driven evaluation system to assess the sustainable tourism development of drama-related intangible cultural heritage in China. Utilizing the Saaty 1-9 scale and hierarchical analysis method, 52 experts determined indicator weights and current development levels. Four dimensions are evaluated: humanistic value, project quality, tourism development, and sustainability. Results reveal humanistic value as most vital at 41.70 % weight. Secondary factors are project quality (29.89 %), tourism development (20.87 %), and sustainability (7.54 %). Aesthetic value, dissemination degree, and location conditions proved to the crucial tourism indicators. The ideological value of drama is paramount, alongside visibility and climate factors. The evaluation demonstrated strong preservation of humanistic value but deficiencies in tourism development, especially regarding infrastructure. Key recommendations include balancing preservation, dissemination, and innovation; emphasizing ideological value, visibility, and climate suitability; maintaining humanistic diversity; and improving site infrastructure. Further testing of evaluation indicators across periods is warranted alongside examining green revitalization potential. This assessment, guided by experts, offers a thorough framework for the sustainable development and preservation of the precious intangible heritage embodied in Chinese drama.