Abstract Aiming at the problems of low accuracy and large prediction errors caused by the serious overlap of multi-metal spectral signals in zinc smelting industrial wastewater, a characteristic interval modeling method is proposed. First, according to the absorption spectra of mixed solution, the characteristic intervals of copper and nickel are preliminarily screened by using different partition lengths. Second, take the smallest root mean squares error of cross validation and the largest correlation coefficient as the evaluation indicators, compare the full-spectral model and each local model, and select the optimal feature sub-intervals of copper and nickel. Last, the partial least squares method is used to model the combined wavelengths of the optimal sub-intervals to realize the simultaneous detection of copper and nickel. The linear determination ranges are 0.3–3.0 mg/L for copper and nickel. the correlation coefficients of copper and nickel are 0.9974 and 0.9966, respectively. The results show that the method reduces the complexity of the wavelength variable screening process, improves the accuracy of the model, and lays the foundation for the accurate analysis of polymetallic ions in zinc smelting industrial wastewater. The zinc hydrometallurgy is mainly composed of a series of processes such as roasting, acid leaching, electrowinning and casting . When th−4–10−6/L), which is in line with the concentration of ions in zinc hydrometallurgy industrial wastewater  spectrophotometry, inductively coupled plasma mass spectrometry, liquid chromatography and polarography . Considestewater . 2) Becastewater . 3) The stewater . Howeverstewater , their astewater , and thestewater , resultistewater . Therefostewater .At present, the commonly used wavelength variable selection methods mainly include: interval partial least squares method, Monte Carlo uninformative variable elimination method and competitive adaptive weighting method. However, in the case of serious spectral overlap, the first method rely on experience to define the size of the wavelength division and the number of combination intervals, and the screening process often results in multiple selections or omissions of the characteristic wavelength variables. The latter two methods separately sample and screen each wavelength, the screening process is complex, and the sampling process is random, which will lead to inaccurate calculation of variable stability index, thus affecting the screening results of characteristic wavelength variables. Therefore, this paper proposes an efficient and high-precision characteristic interval modeling method. This method aims to select the optimal characteristic points of each ion to be measured, reduce the blank information, solve the problems of serious spectral masking and the difference of inhibition degree, and improve the model detection accuracy.R2), and the two calculation formulas are as follows:ith sample, ith sample, R2 is to 1, the higher the accuracy of the model. Therefore, by comparing the RMSECV and R2 values of each sub interval of each metal ion to be measured, the optimal sub interval set is selected. The basic steps of the algorithm are:R2 values of Cu and Ni are calculated according to (1) The global PLS model of Cu and Ni are established in the full spectral range, and the RMSECV and R2 values of copper and nickel in P sub-interval.(2) Set P (5 < P < 15) cycles, divide the full spectra into P equal width sub intervals, establish the local PLS model of copper and nickel on each sub-interval, and calculate the RMSECV and R2 values of the full spectral model and each local model, remove the wavelength interval in which RMSECV is greater than the global RMSECV and R2 is less than the global R2, and take out the remaining Q sub-interval with smaller RMSECV and larger R2 value.(3) Compare the RMSECV and R2 values of all possible combination intervals.(4) Combine t (1 ≤ t ≤ Q) intervals in the remaining Q sub-intervals for PLS modeling, and calculate the RMSECV and R2 as the evaluation criteria of each combination interval, the interval combination corresponding to the minimum RMSECV and the maximum R2 are selected as the optimal interval combination. The algorithm flow chart is shown in (5) Similarly, taking the values of RMSECV and The characteristic interval modeling method is a method for screening the characteristic wavelength variables with high contribution to the model to improve the accuracy of the model. This method performs overall and regional screening of the characteristic wavebands of each metal ion to be measured, avoids the instability of the random sampling process of a single wavelength, and reduces the complexity of the wavelength variable screening process. At the same time, through the evaluation index of local interval, the bands with serious spectral overlap of metal ions are effectively avoided, and the characteristic bands of metal ions are selected. The measurement indicators of characteristic interval variables are root mean squares error of cross validation (RMSECV) and correlation coefficient . The T9 spectrophotometer is a high-sensitivity scientific grade UV-Vis spectrophotometer, equipped with a high-performance PMT receiver and a special grating, mixed C-T dual monochromator and dual beam, so it has a high dynamic range and good SNR. The hexadecyl trimethyl ammonium bromide was used as stabilizer solution, nitroso R salt solution (0.4%) was chosen as chromogenic reagent, acetic acid-sodium acetate buffer (pH = 5.5) was prepared as buffer solution, and standard stock solutions of copper (12.5 mg/L) and nickel (12.5 mg/L) were prepared.First, transfer the appropriate amount of copper and nickel standard solutions to a 25 ml volumetric flask. Then, add 2 ml nitroso R salt, 3 ml hexadecyl trimethyl ammonium bromide and 5 ml acetic acid-sodium acetate buffer. Finally, dilute and mix the solution with distilled water to ensure uniformity. The ultraviolet visible absorption spectra of the solution is measured with a reagent blank in the wavelength range of 350–600 nm. The prepared copper and nickel concentration ranges are both 0.3–3 mg/L 60 groups of copper and nickel mixed solutions in different proportions were prepared for spectral modeling analysis.The spectral signal of copper and nickel single ion solution and the mixed solution are shown in According to R2) are shown in To evaluate the linearity of copper and nickel in the zinc wastewater solution, multiple wavelengths were randomly selected to perform linear fitting of absorbance and concentration. The linear regression equation and correlation coefficient (R2) is used to characterize the linear correlation of ions. The larger the value of R2, the better the linearity. It can be seen from R2) at wavelengths of 375 nm, so the linearity is very poor; it has good linearity at 417, 450 and 496 nm. Because the effective information of copper and nickel at different wavelength points is different, when modeling and detecting the concentration of copper and nickel by the partial least squares method, it is necessary to select the characteristic wavelength variables, and select the wavelength variables with high spectral signal sensitivity, large amount of information and strong linear correlation, so as to improve the accuracy of simultaneous detection of copper and nickel.The correlation coefficient  characteristic intervals are combined in turn in the remaining Q sub intervals to establish a joint interval PLS model. The combined modeling of the optimal characteristic intervals of copper and nickel after each interval division is shown in According to the steps of characteristic interval wavelength selection method described in R2 is 0.9978. The optimal feature interval combination of nickel is to divide the whole band into 12 intervals, and when the fourth, sixth and seventh intervals are combined in sequence, the minimum value of RMSECV is 0.0459 and the maximum value of R2 is 0.9965. The results of the combined wavelength screening of characteristic intervals are shown in It can be seen from The proposed characteristic interval modeling method (CIM) is used to detect the concentration of copper and nickel mixed solutions in 30 groups of prediction sets, and the performance is compared with full band partial least squares method (FBPLS), competitive adaptive weighting method (CARS) and Monte Carlo uninformative variable elimination method (MC_UVE). R2) is higher. The average relative errors of Cu and Ni in the 30 validation sets are 3.27% and 4.12%. The maximum error of Cu is 7.16%, and the maximum error of Ni is 8.33%. The relative error is within 5%, which meets the requirements of industrial detection. As can be seen from R2) of Cu and Ni are 0.9974 and 0.9966, respectively. The results indicate that the proposed characteristic interval modeling method can effectively realize the simultaneous detection of copper and nickel in zinc industrial wastewater.It can be seen from A variety of heavy metal ions coexist in zinc industrial wastewater. Their chemical properties are similar, their mutual influence is serious, and their absorption spectra overlap severely. The full spectra information contains a lot of redundant non-characteristic information, which leads to low accuracy of analysis models and large prediction errors. Therefore, the traditional multi-variable linear analysis method of modeling the full-band information is difficult to achieve accurate detection of polymetallic ions in zinc smelting wastewater. Aiming at the problems of low accuracy and large prediction errors caused by the serious overlap of multi-metal spectral signals in zinc smelting industrial wastewater, a characteristic interval modeling method is proposed. This method screens wavelength variables in a coordinated and partitioned manner, avoids overlapping serious bands, ensures the stability of the screening process, reduces the complexity of wavelength screening, and improves the accuracy of the model. The proposed method is successfully used to analyze the severely overlapping spectra of Cu and Ni. The results show that the characteristic interval modeling method has obvious advantages in the separation of overlapping spectra compared with other methods. The proposed method lays a solid foundation for the accurate analysis of polymetallic ions in zinc smelting industrial wastewater.


Oral mucositis (OM) is a painful lesion that takes place in the mucosa of the oral cavity, usually its etiology is associated with drug therapies in cancer patients. It is presented as well-defined ulcers whose painful symptomatology sometimes implies the suspension of oncological treatment or parenteral feeding, being therefore an important adverse effect, marking the evolution of these types of therapies against cancer. The present work aim is to know the prevalence of oral mucositis in oral cancer immunotherapy compared to its prevalence in standard therapy. A protocol was developed for a systematic review following PRISMA® guidelines and a focused question (PICO) was constructed. A comprehensive literature search was conducted on electronic databases including PubMed, the SCOPUS database, the Cochrane library and the Web of Science (WOS). Six clinical trials were included that met the different inclusion criteria. In these articles, a discrepancy between the prevalence of OM in patients treated with chemotherapy and patients treated with immunotherapy related to the immune checkpoint PD-1/PD-L1 (Nivolumab and Pembrolizumab) was observed. The prevalence of oral mucositis is lower in new immunotherapy with monoclonal antibodies against oral cancer than drugs used so far . However, more studies should be carried out to confirm these data. Key words:Oral mucositis, PD1, PD-L1, oral cancer, immunotherapy. Cancer represents a disease with a high mortality rate, corresponding to one of the leading causes of death worldwide . Oral sqMost patients with OSCC have a poor prognosis due to the late diagnosis of the disease, in which surgical or pharmacological treatment with chemotherapy is limited and with negative results, with a life expectancy of 5 years in 50% of the cases after diagnosis. Besides, unfortunately, the average survival of patients with disease recurrence or metastasis is around 8-10 months .Recent studies indicate that immunotherapy is amongst the most promising strategies in modern cancer therapy. It aims to strengthen innate and adaptive immunity, highly specific for tumor cells, and with low toxicity to healthy host cells -6.Cancer triggers an innate and adaptive immune response of the host during its evolution. Immune tolerance to the tumor is recognized as one of the main characteristics of cancer . Cancer The expression of PD1 and PD-L1 in neoplasms has been shown to be relevant in clinical trials with monoclonal antibodies used for their treatment. In head and neck cancer, the development of clinical trials to understand the effect of immunotherapy on the tumor is thriving, and currently, two monoclonal antibodies have been approved for the treatment of this cancer (Nivolumab and Pembrolizumab) that have been used in studies of patients with advanced, recurrent disease and resistant to chemotherapy, allowing their comparative study -6.- Standard therapy and chemotherapyFor oral cancer, the first-line treatment modality is surgery. Unlike other types of cancer where adjuvant treatment such as chemotherapy is given after surgery, this does not take place in the case of oral cancer and chemotherapy is reserved for more advanced carcinomas, depending on their location, size and risk of recurrence ,7.The two primary modalities with curative potential in the OSCC are surgery and radiotherapy. In the case of chemotherapy, no promising results have been obtained and its use is mainly as palliative treatment ,7. AmongA more recent drug in standard therapy, used in combination with radiotherapy, is cetuximab, a monoclonal antibody directed against the epidermal growth factor receptor, which has an established role in the primary treatment of advanced and unfavorably located oral cancer ,11. Howe- Oral MucositisCancer treatment usually produces a series of changes on normal cells. The gastrointestinal mucosa, especially the oral mucosa, are very susceptible to both direct and indirect toxic effects from chemotherapy and radiotherapy. In the oral cavity, this risk is the result of several factors, such as the high rate of mucosal cell renewal, the existence of a complex and diverse microflora, and trauma to oral tissues during normal oral function -15.Mucositis is the painful inflammation and ulceration of the mucous membranes that line the digestive tract, commonly developing secondary to cancer treatment with radiotherapy or chemotherapy. Mucositis can occur at any level in the gastrointestinal tract, although it is more common in the mouth, receiving the name of oral mucositis (OM) -16.The World Health Organization (WHO) developed a classification of mucositis, being the mildest form the one that has an erythema with non-specific discomfort, burning sensation and hypersensitivity to food (Grade 1). Grade 2 also causes extensive ulcers and mild pain, and food may still be swallowed. Grade 3 increases the severity of the ulcers and therefore the pain, there is difficulty in speaking and the possibility of intake is limited to liquids. The final grade in this classification, grade 4, is characterized by very extensive ulcers, hyposialia, and very significant pain that makes swallowing and even fluid intake impossible ,18.The intensity of the antineoplastic treatment conditions, to a great extent, the appearance of the oral mucosa's adverse effects. The occurrence of OM can be as high as 100% in patients with chemotherapeutic treatments and with radiation in the head and neck region. Patients also report that this is the most uncomforTable side effect they experience from antineoplastic treatment .The present paper aims is to determine the prevalence of oral mucositis in immunotherapy of the immune checkpoint PD1/PD-L1 against cancer compared to its prevalence in standard therapy.P=patient/problem/population; I=intervention; C=comparison; O=outcome) was constructed, taking into account different headings corresponding to the study population, the intervention, the comparison and the outcome. In this case, the question asked is the following:A protocol was developed to conduct a systematic review following the PRISMA® Statement guidelines  ,20. Base“Is the prevalence of OM in patients with head and neck cancer treated with immunotherapy lower compared to the prevalence of OM in those treated with chemotherapy drugs?”- Inclusion criteriaTypes of studies: (a) Original clinical trial-type articles published in scientific journals; (b) studies carried out on human beings; and (c) studies in the English language.Type of population: Patients undergoing treatment with FDA-approved immunotherapy drugs for the treatment of head and neck cancer  or undergoing approval currently being evaluated in clinical trials such as the monoclonal antibody Durvalumab. Studies should reflect adverse effects and, if possible, include a placebo group or be compared with a group with standard therapy used in head and neck cancer.- Exclusion criteriaExcluded studies have the following characteristics: (a) Those studies that did not reflect the prevalence of mucositis in the compared groups; (b) Those articles whose study base focused on other neoplasms did not include the head and neck region; (c) literature review studies, case reports, letters to the editor, abstracts or conference papers; and (d) Those published in a non-English language.No time period was set for inclusion and exclusion criteria as the less recent articles were published in 2015.- Sources of information usedA comprehensive search of the literature was conducted in electronic databases, including PubMed , the SCOPUS database, the Cochrane library, and the Web of Science (WOS) on April 21, 2019.- Search StrategyThe search strategy was carried out using the following terms linked through the following Boolean algorithms: "Head and Neck Neoplasms" AND .- Bias risk assessmentBias risk assessment was assessed independently by two reviewers. Disagreements were resolved by consensus between the two reviewers or the intervention of a third author.Using the predetermined 10 domains for the methodological quality assessment according to the Joanna Briggs Institute Prevalence Critical Appraisal Tool , we deteThe initial search showed a total of 607 articles . After applying the first inclusion criterion of clinical trials in PubMed and the inclusion of scientific articles in the rest of the database, the result was reduced to 262 articles  . By eliminating the duplicated articles from the different databases, a total of 123 articles remained to be read, since 126 were duplicates. The title and abstract of the 123 articles were read and the flowchart for possible inclusion or exclusion was established. The reasons for exclusion of a total of 96 articles after reading the title and summary can be seen in the appendix to this document. A total of 27 articles were read entirely to determine the articles' final inclusion in this review. In this case, 21 articles were excluded for the reasons reflected in the appendix to this document and in the flow chart, thus only 6 articles were included for the elaboration of the results of this work.The data of these 6 articles are summarized in et al. [2016] present an analysis of the CheckMate 141 clinical trial, a randomized phase III trial in which Nivolumab is studied against the investigator's chosen therapy  in patients with recurrent metastatic and platinum-resistant head and neck carcinoma after 6 months of treatment , of which 13.1% of the total sample were grade 3 or 4 adverse events that occurred in the nivolumab group, but these grade 3 or 4 adverse events occurred in 35.1% in the standard therapy group .Data on the presence of OM were reflected in this study, corresponding to 5 cases (2.1%) in the nivolumab group and 10 cases (9%) in the standard therapy group .et al. [2019] present a phase III clinical trial and in their article, they reflect the efficacy and safety of the drug pembrolizumab compared to the standard therapy. This trial involved 97 medical centers in 20 countries. The patients included in the trial were platinum-resistant subjects who did not respond to therapy after 6 months of treatment. Two groups were used, a pembrolizumab group at 200 mg dose every 3 weeks intravenously compared to a group with a standard therapy chosen by the investigator  (Cohen tuximab) .A total of 247 patients received pembrolizumab and 248 received the standard therapy . The mean age in the pembrolizumab group was 60 years [55-66], compared to the standard group which was 60 years (range from 54 to 66 years) .The overall survival of the treated population was 8.4 months in the pembrolizumab compared to 6.9 months in the standard group. There were fewer adverse effects with pembrolizumab in comparison to the standard therapy, finding 33 cases with pembrolizumab (13%) versus 85 cases (36%) in the standard .The authors indicate that the most common adverse effect with pembrolizumab is the development of hypothyroidism in 33 patients . The most common adverse effect in standard therapy is fatigue in 43 patients  .In this article the authors report a prevalence of OM in 6 cases (2%) in the pembrolizumab group and a case of mucositis of a higher grade than 3. While in the standard therapy group, 28 patients had OM (12%) and in 11 of them (5%), it was of a higher grade than 3 .et al. [2017] analysed the sample of Asian subjects from the CheckMate 141 clinical trial. This sample has a total of 34 patients, 23 in the nivolumab group and 11 patients in the standard therapy group   and 10 of the standard therapy group (90.9%). Of these adverse effects, grade III or greater occurred in 2 patients in the nivolumab group (8.7%) and 3 in standard therapy (27.3%) .Regarding the presence of OM, in the nivolumab group it occurred in 1 case (4.3%), while in the standard therapy it occurred in 3 cases (27.3%) .et al. [2017] is the KEYNOTE-012 clinical trial, in phase I, so there is no control group sample. The drug studied in this trial is Pembrolizumab at a dose of 300 mg every 3 weeks. Twenty-six patients with an average age of 62 years participated and, in this study, 62% of the patients presented adverse effects, finding 1 case (4%) of OM (Tahara %) of OM .et al. [2016] show the results of the KEYNOTE-012 clinical trial in phase I-B. In this trial they used pembrolizumab at a dose of 200 mg intravenously for 3 weeks in a total of 132 subjects with an average age of 60 years. A total of 82 cases (62%) presented adverse effects, finding a total of 8 cases (6%) with a grade greater than 3 and a prevalence of OM in 2 cases , 24 (13%) with a degree greater than 3 and a total of 4 cases of OM (2%)  . In thisIn order to answer the objective of this work, the review was carried out with special attention to the clinical trials published to date. In these articles a discrepancy between the prevalence of OM in patients treated with chemotherapy and patients treated with immunotherapy (Nivolumab and Pembrolizumab) has been observed.et al. . This creates a lack of information in this regard but despite these limitations, it is possible to extract from this review that the prevalence of OM in new oral cancer immunotherapy drugs is significantly lower than in standard therapy used to date.et al. [2017] should be highlighted . However, further studies should be carried out to confirm these data.


On the contrary, the operative time of Group 1 was reduced significantly (p = 0.001), with an average of 20% with regard to in vivo procedures performed without rehearsals (on the control Group 2). Conclusions: The study demonstrated that the use of 3D-printed models can be beneficial to dental surgeon practitioners, as well as to students who must be trained before performing clinical treatments.Background and Objectives: The aim of this study is to demonstrate the increased efficiency achieved by dental practitioners when carrying out an ex vivo training process on 3D-printed maxillaries before performing in vivo surgery. Materials and Methods: This developed ex vivo procedure comprises the following phases: (i) scanning the area of interest for surgery; (ii) obtaining a 3D virtual model of this area using Cone Beam Computed Tomography (CBCT); (iii) obtaining a 3D-printed model , on which (iv) the dental practitioner simulates/rehearses ex vivo (most of) the surgery protocol; (v) assess with a new CBCT the 3D model after simulation. The technical steps of sinus augmentation and implant insertion could be performed on the corresponding 3D-printed hemi-maxillaries prior to the real in vivo surgery. Two study groups were considered, with forty patients divided as follows: Group 1 comprises twenty patients on which the developed simulation and rehearsal procedure was applied; Group 2 is a control one which comprises twenty patients on which similar surgery was performed without this procedure . Results: Following the ex vivo training/rehearsal, an optimal surgery protocol was developed for each considered case. The results of the surgery on patients were compared with the results obtained after rehearsals on 3D-printed models. The performed quantitative assessment proved that, using the proposed training procedure, the results of the in vivo surgery are not significantly different ( In dental clinics, patients often have terminal edentulism that cannot be treated using traditional prosthetic methods ,2,3. AlsTo optimize the necessary in vivo procedures, extra support can be provided by three-dimensional (3D) printing. The scope of using this technique is to be able to rehearse, ex vivo, the surgical maneuvers before performing them on the patient. Therefore, the expected rate of success in vivo may increase, as well as the precision and time-efficiency of the intervention. In this respect, craniofacial surgery represents one of the top areas where 3D printing technology has built up interest ,11,12. TIn general, 3D printing, also known as rapid prototyping, is a process of forming a solid object of a required shape. It was first described and patented in 1986 by Charles Hull  and in 1In the present work, maxilla models are obtained through 3D printing using Computer-Aided Design/Computer-Aided Manufacturing (CAD/CAM). The design of the 3D-printed models is based on the clinical data obtained from intraoral scanning, as well as on paraclinical data retrieved from the patients using Cone Beam Computed Tomography (CBCT). The obtained 3D-printed models are utilized for ex vivo training for implant insertion and sinus lift simulations before performing the real, in vivo surgery in the clinical area.The first hypothesis of the study is to prove that using such implant simulations and clinical procedure rehearsals provides a superior quality of prosthetic restorative treatments on dental implants. To verify this hypothesis, a comparison must be made between the clinical results obtained in the final in vivo procedure and those obtained when the ex vivo training on models is performed.The second hypothesis is that intervention time is reduced when such simulations are performed, compared to the situation when they are not utilized. To verify this hypothesis, a comparison must be made between the operative time necessary for the final in vivo procedure when such simulations (and the corresponding ex vivo training on models) are performed and the operative time necessary without this developed procedure. Forty patients were included in this study. They were divided into two groups: Group 1 comprised twenty patients selected for the developed clinical procedure, which included simulations/rehearsals, as described in the following for one of these clinical cases. Similar procedures were performed on each of these twenty patients. Group 2 was a control group, consisting of other twenty treated patients on which in vivo surgery was performed without the training developed in this work. This group was considered in order to evaluate the second hypothesis of this study. As a necessary remark, the cases of Group 2 were selected from a larger group of patients on which the type of surgery specific to this study was performed previously to developing the simulation and rehearsal procedure developed in this work. The selection was made by considering an as good as possible match (in pairs of patients) with the cases of Group 1 from the point of view of their clinical difficulty . The initial 3D design of the maxillary ready for printing , was employed to obtain the 3D polymeric model of the hemi-maxilla for each considered case/patient. The Digital 3D Printer Form 2 in In this third step of the procedure, the dental practitioner performed on the polymeric 3D-printed hemi-maxilla model the simulation of the sinus lift procedure by lateral approach, the insertion of the ex-vivo implant, as well as the later and crestal augmentation. Thus, the surgeon was able to observe the exact positioning of the bone formation, the thickness of the remaining bone, and the distance to the sinus membrane. As a necessary remark, the following operative phases described in the following section could not be rehearsed on the 3D-printed model: incision, flap elevation, sinus membrane elevation, flap repositioning, and suture.The real operation was performed between 2 to 5 days (maximum) after the rehearsal, depending on the availability of the operation room and the surgeon. The essential criterium was to have the same healthy sinus condition for the patient.In this fourth step of the procedure, correlating clinical information with preclinical data, as well as with those obtained on 3D-printed models, the medical doctor was able to pass to the surgical procedure for the sinus lift and the insertion of dental implants, as shown in the example in The first clinical stage was the sinus lift. For this case, a sinus floor augmentation procedure via the lateral window approach with simultaneous implant placement for the missing 16 and a sinus augmentation for latter implantation for 17 were planned. On the day of the surgery, this patient received 2 g of amoxicillin one hour before the procedure as antibiotic prophylaxis: 1 g one hour before the procedure and the rest of the dose 12 h after the first dose. As post-operative therapy, the patient received 1 g three times a day per os for 7 days . FollowiA bony window was then traced in the 16 and 17 regions using a piezo-surgical unit b,c. The ® B., Dentegris GmbH, Monheim am Rhein, Germany). This specific augmentative material was chosen because of its mineral composition, physical, chemical, and biological properties, as well as because of its hydrophilic surface, with a close resemblance to the characteristics of the human bone. Also, this material allows for fast revascularization because of its interconnecting pore system. Thus, it was able to provide long-term stability of the augmented site with predictable results [A C1 Implant 3.75 mm × 11.5 mm Standard Platform Conical Connection  with simultaneous sinus lift procedure was inserted on the 16 position e. A dedi results ,25.® Membrane, Dentegris GmbH, Monheim am Rhein, Germany). At the level of 17, a sinus elevation procedure with a simultaneous lateral and vertical augmentation of the crest was performed f. The auThe incisions were closed with interrupted sutures 4-0 polyester .A CBCT was performed the day after the surgery in the fifth step of the procedure. Thus, each patient was followed up two weeks after the surgery to check if the healing was uneventful. Also, no complications occurred in the six-month period during the osteointegration of implants.For the considered case, this CBCT assessment is presented in the cross-sections in All the phases above were performed, for each patient in the study, by the same surgeon. All the CBCT examinations were performed by the same radiologist.Following the described procedure, a comparison was made between the mezio-distal positions of the implant on the maxillary arch on the CBCT of the model and the patient’s postoperative CBCT, as shown in the panoramic views in D1-3D and D1-P) were done with the non-parametric Wilcoxon Signed Ranks test (p < 0.05 (p = 0.089).The statistical processing of these results was performed using SPSSv17, calculating the mean and the standard deviations (SD). Descriptive statistics were performed for the numerical variables and comparisons between these series of pair values  and when performing implant procedures on patients (D2-P). Descriptive statistics were performed for the numerical variables. Comparisons between these series of pair values were done with the non-parametric Wilcoxon Signed Ranks test (p < 0.05 (p = 0.350).Similar statistical processing was performed using SPSSv17 for the distance from the ridge margin to sinus window, in a comparative look between the data obtained when rehearsing on 3D-printed models . The comparison between these time intervals for each of the twenty patients considered in the control Group 1 and in the study Group 2 was made in Another important aspect of a surgical procedure, essential from the patient’s point of view, is the operative time. We registered the corresponding operative time intervals T for the procedures described above for the patients in Group 1  and compared them to the time intervals 0 to 200 in The twenty clinical cases of the control Group 2 (numbered 10T), i.e., for Group 2, and when implant procedures on patients were performed after such a rehearsal on 3D-printed models (T), i.e., for Group 1. Descriptive statistics were performed for the numerical variables and comparisons between these series of pair values using the non-parametric Wilcoxon Signed Ranks test  was 117.55 min. The mean time meanT needed for the same procedure in the cases of the patients of Group 1  was 93.2 min. The 24.35 min difference between these two values, as well as the relative decrease (0 meanT − meanT)/0 meanT equal to 20.71% from the 0T surgery time indicates the benefit of applying the rehearsal procedure for patients.The mean time p < 0.05 (p < 0.001).The results were considered significant for a value of p < 0.05 . The difSurgeons often use different 3D-printed human organs before surgery, for example in craniofacial reconstruction ,27 and iWith 3D printing, dental medicine can also evolve in terms of the quality of future implant surgical treatments, and this has been approached in numerous studies ,32,33,34On the other hand, oral implantology is not a stand-alone branch; it depends equally on the future prosthetic treatment that must be consistent with the insertion of implants . Tissue The present study approached the utility of the 3D-printed models in the visualization of the complex maxillary anatomical structures and the impact of the surgery simulation before the in vivo operative time. Both maxillary implant insertion combined with sinus augmentation procedures require an accurate technique. Guidance is necessary to achieve the appropriate angulation and position in relation to the adjacent teeth, as well as a preplanned design and positioning of the lateral window for approaching the sinus. Due to the realistic form of the 3D-printed models, the technical steps of sinus augmentation and implant insertion could be performed on the corresponding model prior to surgery. Technical steps of the procedure were rehearsed in the simulation using the 3D model : shape and size of antrostomy, implant insertion, and grafting. As pointed out, several operative phases could not be rehearsed on the 3D-printed model: incision, flap elevation, sinus membrane elevation, flap repositioning, and suture .In this work, we analyzed the compliance of the ideal mezio-distal position of the implant due to its crucial role for the final prosthetic restoration and the health of the tissue around it . The staAs an overall result, the statistical data showed no significant differences between the 3D-printed model and postoperative CBCTs.The first hypothesis of the study was proved: using implant simulations allows for a superior quality of future prosthetic restorative treatments on dental implants. The second hypothesis was also confirmed: a significant reduction of the intervention time of around 20% was obtained  when rehIn the complex clinical cases approached, the 3D-printed models allowed for a precise plan of the surgery. Also, the transfer of the accuracy in the real operation field was made possible. Therefore, using 3D-printed models, implants can be inserted in vivo in full compliance with future prosthetic treatment. Improving the surgical operative time, as obtained in this study, is a real benefit to both the patient and the surgeon. An accurate sinus lift procedure along with reduced tissue manipulation leads to a better perception during the recovery time.In conclusion, the use of 3D-printed models can benefit implant practitioners, as well as students taking a course in Oral Implantology. Also, it can be a support to students of dental medicine in implantology to simulate implant insertions. Thus, 3D-printed models can play a significant role in the educational field. The anatomical models mitigate the inconvenience of cadaveric dissection, allowing the student to focus only on the structural anatomy and to relate it with its functional role . In contWe expect that further progress in the field will lead to the development of both soft and hard printable materials that mimic the haptics of biological tissues. The cost-effective availability of 3D printers may allow practitioners to enhance their surgical skills on 3D-printed models, with a resolution similar to one of the original dissectible structures. For the imaging part, future work in this direction in our group includes using combined Optical Coherence Tomography (OCT) ,49,50 an


AFM images reveal that the number of NP clusters interacting with the lysozyme fibers does not increase significantly with NP volume concentration, suggesting a saturation in NP aggregation on the fibrillary surface. The data indicate that the number of non-adsorbed Fe3O4 NPs is highly dependent on the timing of NP infusion within the synthesis process. SAXS data yield access to the spatial distribution, aggregation manner and density of NP clusters on the fibrillary surfaces. Employing modern data analysis approaches, the shape and internal structural morphology of the so formed nanocomposites are revealed. The combined experimental approach suggests that while Fe3O4 NPs infusion does not prevent the fibril-formation, the variation of NP concentration and size at different stages of the fibrillization process can impose a pronounced impact on the superficial and internal structural morphologies of these nanocomposites. These findings may be applicable in devising advanced therapeutic treatments for neurodegenerative diseases and designing novel bio-inorganic magnetic devices. Our results further demonstrate that modern X-ray methods give access to the structure of—and insight into the formation process of—biological–inorganic hybrid structures in solution.Understanding the formation process and the spatial distribution of nanoparticle (NP) clusters on amyloid fibrils is an essential step for the development of NP-based methods to inhibit aggregation of amyloidal proteins or reverse the assembling trend of the proto-fibrillary complexes that prompts pathogenesis of neuro degeneration. For this, a detailed structural determination of the diverse hybrid assemblies that are forming is needed, which can be achieved by advanced X-ray scattering techniques. Using a combined solution small angle X-ray scattering (SAXS) and atomic force microscopy (AFM) approach, this study investigates the intrinsic trends of the interaction between lysozyme amyloid fibrils (LAFs) and Fe The progressive deposition of amyloidal fibrils over a long period in one’s life time was postulated as a major cause of some chronic life-threatening diseases such as type II diabetes , atheros3O4), gold (Au) and zinc oxide (ZnO) nanoparticles (NP), to name just a few, received significant attention from the research community due to its unique biochemical and physical properties, and was verified to be instrumental in facilitating various magnitudes and manners of interaction with amyloidal β (Aβ) fibrils [Nanomedicine that comprises nanomaterials such as iron oxides , in combination with further experimental methods, allows the study of the structural morphology of such nanocomposites.The process of nanoparticles’ adsorption on the protein surface can be influenced by several factors. More recently, in our previous work , the str3O4 NPs, and thus to elucidate the manner and morphology of the aggregates of NP clusters on the fibrillary surface. The understanding of such NP–protein interactions is the first step towards deciphering the true nature of the NP-mediated biological and biomedical effects and might lead to strategic manipulation in vivo. Our study demonstrates how modern solution SAXS approaches can reveal such structural information on biomaterials.Following our previously established approach, the aim of the present work was to elucidate the influence of nanoparticle size and concentration on the formation of fibril-nanoparticles complexes and its dependence on the infusion time. Using nanoparticles of different sizes and concentrations, the distribution patterns, clustering manner and interactive morphology of the resulting fibrillary nanocomplexes were studied by solution SAXS and AFM measurements. While the SAXS data allowed the characterization of the shape and internal structural morphology of the nanocomposites and yielded structural models of these, AFM images were taken to ensure mutual interactions between LAFs and FeMutual interaction between the NPs and LAFs, and thus, the manner of NP adsorption on the fibrillary surface of the pure and composite fibrillary samples were determined by AFM imaging. AFM images of pure LAFs at different magnifications are shown in To study the dependence of the fibrillary interaction with 10 nm NP on volume concentration, 10 μL and 30 μL of the NP solution were infused separately with 300 μL of LAFs, and acquired with AFM imaging as shown in The figures at the highest resolution show NPs adsorbed on the LAFs, confirming the binding affinity between the fibrils and 10 nm NPs. These results are in accord with previous studies, which reported such an attachment of NPs on the fibril surface ,24,25,26When 10 nm NPs were added to the solution before the fibrillization process was started (type-B process), LAF formation was observed as well as NP adsorption. The morphology of the aggregated NP clusters on the fibrillary surface, shown in By carrying out the fibrillary interaction with 20 nm NPs before the fibrillization (type-B process) , one can3O4 NPs of 10 nm, 20 nm and 30 nm , are opposite and similar in the range of magnitude, the numbers of un-adsorbed NP clusters are proportionally higher as the concentration increases, implying a limiting saturation on the number of NP clusters on the fibrillary surface. In addition, a noticeable difference is the association of less floating NPs with the type-A process as opposed to the type-B process.Subsequently, the size of the NPs was increased to 30 nm. q = 0.9 nm−1. In addition, the corresponding pair–distance distribution function p(r) of the pure LAFs (presented in maxD = (80 ± 2) nm; however, due to the limited resolution of the SAXS instrument, the larger species, which are likely present in solution, cannot be resolved properly.Following the 2D morphological characterization of the nanocomposites by AFM imaging, SAXS data were collected for the same sets of freshly prepared samples in solution. In the first step, the SAXS profiles of the pure species, LAF and NPs of various sizes, were studied and are shown in p(r) function reflects the elongated shape of the fibrils with an effective cross-sectional diameter of d = (23 ± 2) nm. From the SAXS data, an ab initio shape model of the average solution structure of LAF was determined using the ATSAS program DAMMIF  for polydisperse spheres was computed with GNOM  fr of d =  ± 2 nm. ith GNOM c, from wq-range from 0.05 nm−1 to 1.0 nm−1. In general, with increasing of the NP concentration, the contribution of the P20 form factor significantly contributes to the scattering signal, most prominently seen by the presence of the characteristic oscillations. The significant signal of P20 at a rather low mixing ratio of 10/300 (NP/LAF) for samples B10 and A10 stems from the much stronger scattering power of Fe3O4 compared to the weaker scattering by lysozyme protein (~100 times less), which contributes at smaller scattering angles [q < 0.2 nm−1), while the rest of the curves look similar. In order to analyze how significantly the scattering curves from the simple mixture of both species in solution differed, all profiles were fitted as a linear combination of the pure LAF and P20 data using the ATSAS program OLIGOMER [With the species of pure materials being characterized, the fibrillary nanocomposites made of LAFs and the NPs were examined to investigate the morphological effects of varying sizes and volume concentrations of NPs and types of mixing processes. g angles . A direcq < 0.15 nm−1. In particular, the experimental data exhibit a weak but visible shoulder in this angular range, indicating the presence of an additional structural factor contribution. Such an interference signal might imply the formation of clusters or the interactions between the fibrils and the nanoparticles [For the samples made through the type-A process, the model of an oligomeric mixture fits the two lowest concentrations  well, while for A300, there are slight deviations present for articles ,33,34,35q as well as the presence of the contribution of a rather broad structure factor.Likewise, for the samples made through the type-B process, deviations from the simple mixture model are present for all concentrations. For the low concentrations , the curves exhibit a steeper decay, indicating the formation of larger species. The curve for sample B300 exhibits both a stronger scattering intensity at small q-values. For P30, small deviations are present for both series, but to a lesser extent. While the modelling indicates the presence of additional structures in solution, it also shows that LAFs are formed for both processes. Thus, consistent with the AFM results, the time of infusion does not inhibit the fibrillization.For the mixtures of LAF with P10 and P30 b, only tIn order to determine how significantly the experimental SAXS data deviated from a simple superposition of pure LAF and NPs, an alternative analysis was performed to verify how many independent components could be extracted from the sets of SAXS curves shown in q-values.q likely contribute to the observed weak structure factor-like contribution in the experimental data. Note that for the A-series of P10, the DAMMIX analysis yielded a third component with zero weight, consistent with the previous finding of the OLIGOMER analysis that the data are well described as a simple superposition of LAF and P10 without any significant intermediates formed. The structural differences are more obvious when comparing the corresponding p(r) functions. In this real-space representation, one can clearly distinguish the elongated character for all type-B samples (solid lines). These appear as the formation of elongated assemblies of the respective NPs with an overall length of 80 nm, which corresponds to the maxD of LAF. Given the stronger scattering strength of the NPs than the fibril, the data suggest that the evolving species for all the type-B-processes are NP adsorbed or clustered on the fibril axis. For the type-A process, the findings are more heterogeneous. For P10, no intermediate is formed, while for P30 the pair–distance distribution function is very similar to the type-B sample. For P20, the p(r) resembles that of a hollow-sphere with diameter of 70 nm formed by the NPs, implying that it is wrapped around the fibrils, which again, because of the weaker scattering contrast, can be difficult to observe.The computed scattering curves for all samples from the so determined evolving species of the nanocomposites are displayed in Based on the curves for the intermediate clusters shown in The model for the intermediate state of P20A appears as a spherical assembly of ~8 NPs, which, when aligned with the LAF model, suggest adsorption on it from several sides. The model for P30A (not shown) is similar to that for P30B, reflecting their similarity in the scattering curves.Finally, the volume fractions of the three species, LAF, NPs, and intermediate composite as a function of NP/LAF ratio as determined from the DAMMIX analysis are presented in 3O4 NPs of different sizes as well as the role of the mixing time point of NPs during the fibrillization process. It revealed how the formation process can influence the spatial distribution of NP clusters adsorbed on the fibrillary surface and the composition of the final internal structures of the composites. AFM images were acquired to examine the mutual interaction between the NPs and LAFs as well as the aggregating manners of NP clusters on the fibrillary surface and topological patterns of the nanocomposites. Using SAXS measurements and modern data analysis schemes, the presence of nanocomposites in solution were confirmed and their actual solution structures were revealed.The presented study investigated the surface topography and internal structure of the nanocomposites formed by LAF and FeCombining these experimental techniques, specific structural differences in the formation processes of the nanocomposites were determined. Unlike the type-B process, in which the number of un-adsorbed NPs decreased as the particle size increased, very few to null un-adsorbed NPs could be found in the cases of the type-A process. As manifested in the AFM images for NPs of 10 nm, thicker composites were formed than for larger NPs (20 nm and 30 nm). Generally, for both process types, a larger volume concentration of NPs does promote slightly more NP clusters; however, the freely unattached NPs also proportionally increase, suggesting a limited saturation for the integration between the NPs and LAFs. The two significant features of the type-B composites are the higher number of unadsorbed NPs and the larger cluster size than those of type-A composites, which are present for all three NP sizes. Moreover, the type-A process generally produces more NP clusters as the concentration increases, which is not pronounced in the composites by the type-B process. The clusters of 30 nm NPs attached to the LAF appear less abundant and are not significantly larger than those of other NP sizes, which is reasonable due to the fact that the numbers of 10 nm NPs and 20 nm NPs are at least 60 times that of 30 nm NPs.As an alternative to the topographical morphology of the nanocomposites measured by AFM, SAXS measurements allowed the determination of their formation and structure in solution. Based on the detailed analysis, the presence of a fraction of nanocomposites in solution besides pure LAFs and NPs was confirmed. Moreover, ab initio modeling determined the overall average structure of these nanocomposites and their volume fraction as a function of infusion time, mixing ratio and particle size. The resulting models provide insight into the characteristic features of the formed nanocomposites. One noticeable feature of type-A and -B samples is the distinguishable presence of NPs of the respective sizes, and the well-aligned composite structures between the two models in pure and intermediate states. The final morphology of NP agglomeration for the type-B process appears patchy and partially continuous on the entire fibrillary surface, whereas in the type-A process, randomly generated clusters can form on the surface with less adherence to the fibrillary body. This is an assembling morphology that cannot be observed in the topographical presentation of AFM images. In addition, the average number of NPs within these agglomerated clusters increases as the particle size decreases, suggesting a linear relationship between the area of surface coverage and radius of NPs, which was also not clearly elucidated from the AFM analysis on a small portion of the composite solution with a finite number of objects deposited as a thin film.Finally, the volume fractions of pure NPs, pure LAFs and intermediate states were obtained from the modelling. Although the SAXS results did not convey a strong indication that the number and cross-sectional width of NPs clusters are highly dependent on the volume concentration, as suggested by the AFM analysis, such precise estimation of the number of NPs binding on the fibrillary surface is not feasible in AFM imaging considering the randomness of the interaction and its limited view. As it was manifested by the analysis of the SAXS data, the timing of the incorporation of NPs during the synthesizing process of LAFs did affect the morphological formation of the nanocomposites. In other words, the type-A process necessitates higher concentration of NPs before the deviations emerges when compared to the type-B process, of which the deviation appears even at the lowest concentration. In addition, these low-resolution models allow the best depiction of the distribution of compositional components, internal structural morphology and precise manners of NP aggregation in the various types of nanocomposites.3O4 NPs-incorporated LAF nanocomposites and the effects of the NP sizes, volume concentrations and the timing of NP incorporation. The AFM measurements indicate that the addition of NPs of different sizes and concentrations to LAFs before and after the fibrillization process has a profound influence on the morphology and density of the attached clusters on the fibrillary surface. In general, the type-B process produces thicker nanocomposites with small NP sizes than the type-A process. Overall, the higher volume concentration promotes higher abundance of NP clusters alongside an increasing number of freely unattached NPs, which is especially apparent in the nanocomposites made by the type-A process. The SAXS results indicate that the formed NP clusters are highly dependent on the timing of NP infusion but relatively independent of the NP concentration. This study provides an alternative view into the formation of NP-fibril nanocomposites. It reveals a suitable range of NP concentrations to form such composites and underlines the importance of timing for the NP incorporation process. With such parameters, it appears possible to toggle the fibrillary aggregation at different stages of amyloidogenesis, before and after the fibrillization, corresponding to the entities of monomers and protofibrils. Such approaches could be potentially applicable to the treatment neurodegenerative diseases in future studies. Furthermore, such nanocomposites, which are formed under different parameters, may provide an alternative method for synthesizing novel electronic, optic or magnetic devices produced from bio-inorganic compounds [In summary, this combined SAXS and AFM approach provides a complementary perspective to studying the structure of Feompounds ,39,40.3O4 magnetic NPs with the respective hydrodynamic sizes of 10 nm, 20 nm and 30 nm, with 5 mg/mL Fe concentrations and 0.02% NaN3, which were provided by and purchased from Ocean Nano Inc. . These NPs were coated with an amphiphilic polymer coating composed of carboxylic acid atop an organic monomeric layer of oleic acid, and were dispersed in a borate buffer solution of 50 mM with pH of 8.5, and thus, were easily reactive to biomolecules. The sizes of the NPs were uniform within 10% variation and were highly stable within a pH range of 2 to 14, according to the manufacturer’s description.We used FeLyophilized powder of hen egg white lysozyme (HEWL) with protein composition > 90% and molecular weight of 14.3 kDa  was purchased from Sigma-Aldrich Chemical Company ; other chemical reagents for preparation of the buffer solution , from Sigma Aldrich , were all of analytic grade.In brief, the synthetic procedure of LAF solution commenced with the dissolving of the HEWL powder in the buffer solution that consisted of 0.2 M glycine-HCl, with a pH of 2.2 and 80 mM NaCl, settling down at the concentration of 5 mg/mL. The buffer solution was conditioned at the acidic pH level of 2.2 for breaking down of the HEWL powder within a short period of time relative to the naturally occurring process. In the following step, two protocols were used to prepare the nanocomposite samples. To prepare type-A samples, the LAF powder solution was heated for 2 h at 65 °C while being constantly stirred at 250 rounds per minute (rpm) to form LAFs. The prepared LAF solution was doped with NPs of varying sizes and volume concentrations  after the fibrilization process was finished (type-A process). Alternatively, the LAF powder solution was doped with NP of the same size and volume concentration, and was afterwards heated for 2 h at 65 °C while being constantly stirred at 250 rounds per minute (rpm) to form the fibrillary nanocomposites (type-B process).http://gwyddion.net, accessed on 1 January 2021).Atomic Force Microscopy was chosen to study the surface morphology of the pure LAFs and their nanocomposites at nanoscale resolution. The AFM imaging presented in this work was performed using a 5500 AFM system  equipped with PicoView 1.14.3 control software  The images were acquired in tapping mode, using standard rectangular silicone tips with aluminum reflective coating on the back, with a nominal resonant frequency of 300 kHz and a nominal spring constant of 26 N/m . These tips had a visible apex with a very small radius (7–10 nm), ensuring a high lateral resolution to provide images of the fibrils. Samples for AFM observation were prepared by drop casting the LAF solutions on freshly cleaved mica sheets. The sample for AFM, which was actually a dried thin layer of fibrils on mica sheet, was placed on the AFM sample holder. Imaging of all samples was performed in an aerated environment with the humidity in range of 30–40% and at a room temperature of about 26 °C during the measurement. For each sample, a region-of-interest of 1024 × 1024 points was scanned. The captured images were processed using freely available software from Gwyddion  SAXS patterns were recorded using a PILATUS 6 M pixel detector placed at 3.0 m from the samples. With this experimental setting, a total q-range from q = 0.01–7.0 nm−1 could be accessed, with q denoting the scattering wave vector, which is defined as q = 4π/λsin(Θ), and 2Θ denoting the scattering angle.SAXS is sensitive to the structure and interactions of nanometer-sized particles in solution ,32,33,41As all samples exhibited isotropic scattering patterns, the sets of frames for each sample were azimuthally averaged, normalized by the transmitted beam intensity, checked for radiation damage and averaged by the P12 beamline SASFLOW pipeline . The bac3O4 NPs and LAF by combining modern X-ray scattering with AFM. We determined the spatially distributed binding and morphology of NP clusters on the fibrillary surface and the influence of different parameters. The effects of the timing of NP infusion, and the volume concentration and size of NPs, on the internal structural morphology, the aggregating manner of NPs and NP clusters, and the 2D and 3D tomography of the nanocomposites, were studied.We explored the nanometer-scale morphology of nanocomposites formed by Fe3O4 NPs and LAFs is dependent on the particle sizes and NP concentrations. Additionally, average ab initio models of the nanocomposites’ morphology were determined from the experimental SAXS data. Based on these beads-based nanocomposite models, the size, 3D spatial distribution, internal morphology, and manner of the affinity of the NPs on the fibrillary surface were quantified.The 2D surface topography measured by AFM confirmed that the interaction between FeOur study demonstrated how the combination of modern solution SAXS approaches with real-space methods reveals structural information regarding biomaterials. With our combined approach, the adsorptive manner of the NP clusters on the fibrillary surface as a function of particle size, volume concentration, and infusion timing in the fibrillization process were determined. Modeling of the SAXS profiles revealed that the type-B process produces patchy, closely adhering adsorbates of NP clusters on the fibrillary surface, while type-A processes result in a less favorable interaction. For both processes and all conditions studied, LAFs were formed.3O4 NPs are particularly interesting in terms of their potential clinical applications in relation to the accumulation of amyloid fibrils.This work showed that the utilization of the effect of concentration and size on the nanocomposite structure can play an important role in the preparation of magnetically active bio-hybrid systems. Here, the understanding of NP–protein interactions is the first step towards realistic applications of these NP-mediated biological and biomedical effects and even towards strategic manipulations in vivo. While the use of iron oxide NPs in amyloid protein aggregation is still limited, because of their specific advantages, such as low toxicity and high magnetization, which allow for magnetic separation, Fe


This study aimed to investigate the sensory and motor innervation of “Taichong” (LR3) and “Ququan” (LR8) in the rat and provide an insight into the neural relationship between the different acupoints in the same meridian.The LR3 and LR8 were selected as the representative acupoints from the Liver Meridian and examined by using the techniques of regional anatomy and neural tract tracing in this study. For both acupoints, their local nerves were observed with regional anatomy, and their sensory and motor pathways were traced using neural tract tracing with single cholera toxin subunit B (CTB) and dual Alexa Fluor 594/488 conjugates with CTB (AF594/488-CTB).Using the regional anatomy, the branches of the deep peroneal nerve and saphenous nerve were separately found under the LR3 and LR8. Using single CTB, the sensory neurons, transganglionic axon terminals, and motor neurons associated with both LR3 and LR8 were demonstrated on the dorsal root ganglia (DRG), spinal dorsal horn, Clarke’s nucleus, gracile nucleus, and spinal ventral horn corresponding to their own spinal segments and target regions, respectively. Using dual AF594/488-CTB tracing, it was shown that the sensory and motor neurons associated with LR3 were separated from that of LR8.This study demonstrates that LR3 and LR8 are innervated by different peripheral nerves, which originated from or terminated in their corresponding spinal segments and target regions independently through the sensory and motor pathways. These results provide an example for understanding the differential innervation between the different acupoints in the same meridian. The two representative acupoints on the hind limbs namely “Taichong” (LR3) and “Ququan” (LR8) are attributed to the Liver Meridian of Foot-Jueyin in the traditional acupuncture theory . AlthougIn order to reveal the differential innervation between LR3 and LR8 from the PNS to the CNS, their neural characteristics were systematically examined by the techniques of regional anatomy and neural tract tracing on the experimental rats. The two techniques have been used popularly in acupuncture research . Herein,via the neural pathways.With both anatomical and histological approaches, this study focused the observation on the nerves under the regions of LR3 and LR8, and their neural components at the cellular level along the sensory and motor pathways. Besides the traditional consideration of the LR3 and LR8 from the perspective of Meridians, this study is expected to provide the neuroanatomical evidence for explaining the differential innervation of different acupoints in the same meridian This study used nine young adult male Sprague Dawley rats . All the animals were housed in standard cages on a natural light-dark cycle with controlled temperature and humidity, and free access to food and water. The care and experimental manipulation of rats were approved by the Ethics Committee at the Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences (reference number 2021-04-15-1).Six rats were used for the injection of CTB into the LR3 and LR8 on the opposite sides of hindlimbs. Considering the application of the CTB on the peripheral site only produced neural labeling on the same side of injection, bilateral injections were performed in this study. This is also in order to reduce the number of animals used. Under the anesthesia with 2% isoflurane, 2 μl of 1% CTB solution  was injected subcutaneously and muscularly with Hamilton micro-syringe  into the right side of LR3 and the left side of LR8, respectively.The corresponding sites of LR3 and LR8 in the rat were determined according to the principle of comparative anatomy . For theBesides the application of single CTB on the opposite sides of LR3 and LR8, the injection of dual AF594-CTB/AF488-CTB was also carried out on the ipsilateral side of LR3 and LR8 with the other three rats. In this process, 2 μl of 0.1% AF594-CTB and 2 μl of 0.1% AF488-CTB  were separately injected into the LR3 and LR8 on the same hindlimb.Three days after injection, the rats were anesthetized with an overdose of tribromoethanol solution (250 mg/kg) to induce euthanasia and then transcardially perfused with 100 ml of 0.9% physiological saline followed by 250–300 ml 4% paraformaldehyde in 0.1 M phosphate buffer .After perfusion, regional anatomy was performed along the hindlimb to observe the peripheral nerves on the regions of LR3 and LR8 . After oThe transverse sections of the brain and spinal cord and sagittal sections of DRG were cut at a thickness of 40 μm on a freezing microtome . Afterward, these were orderly collected in 0.1 M PB (pH 7.4) with the 12-hole Petri dish.The neural labeling with CTB was stained by immunofluorescence. In brief, the slices were incubated in a blocking solution containing 3% normal donkey serum and 0.5% Triton X-100  in 0.1 M PB for 30 min, then removed to goat anti-CTB  in 0.1 M PB containing 1% normal donkey serum and 0.5% Triton X-100 at 4°C for one night. On the next day, the slices were washed thoroughly with 0.1 M PB and exposed to donkey anti-goat Alexa Fluor 488 secondary antibody   for 1 h. After that, the slices were washed again and then mounted on slides (Superfrost) . The slides were cover slipped with 50% glycerin before the observation.It was noteworthy that the neural labeling with AF594/488-CTB can be directly observed without further staining.The Rat Brain in Stereotaxic Coordinates . Accordingly, the slices with CTB labeling were further observed and recorded using a fluorescent microscope equipped with a digital camera  or a laser scanning confocal microscope . All images were processed using Adobe Photoshop/Illustration CC2017 . The anatomical structure of tissue slices from the brain stem and spinal cord was determined according to rdinates , and theSE and processed with the software GraphPad Prism 8.0.2 . Since there is no distinct boundary between the slices of the spinal cord, the number of labeled motor neurons was counted together without further distinguishing their spinal segments.The number of the labeled neurons was obtained from the six rats with the injection of CTB, in which sensory neurons were counted in all the slices of each DRG, and the motor neurons were counted in every 12th slice of the spinal cord. All the data were expressed as mean ± With regional anatomy, the peripheral nerves associated with LR3 and LR8 were clearly observed under the naked eyes. The common peroneal nerve branches from the sciatic nerve run down along the leg and further gives out the branch of the deep peroneal nerve toward the region of LR3. The saphenous nerve branches from the femoral nerve run into the region of LR8 .The neural components associated with LR3 and LR8 were demonstrated with CTB labeling, including the sensory neurons in DRG, transganglionic axon terminals in the spinal dorsal horn, Clarke’s nucleus, and gracile nucleus, as well as motor neurons in the spinal ventral horn. This labeling is located ipsilaterally corresponding to the side of CTB injection for LR3 and LR8, respectively –4.The sensory neurons for the LR3 were distributed in lumbar (L) 1–6 DRG with higher concentration in L4 DRG , and theThe differential innervation between LR3 and LR8 was further compared with a dual fluorescent tracing technique . Unlike via the sensory and motor pathways. From the PNS to the CNS, it is beneficial for understanding the effective routes of the two acupoints by acupuncture stimulation  interiorly and extend over the body exteriorly, forming a network and linking the tissues and organs into an organic whole. This system included the 12 main meridians running longitudinally and interiorly within the body, with each meridian considered to interconnect with its corresponding visceral organ. However, it is absent until now of structural basis to support this discussed interconnection. In our previous study, we have demonstrated the correlated sensory and sympathetic innervations between acupoint BL 23 and kidney in the rat . In thismulation .The peripheral nerves under the LR3 and LR8 in the rats were demonstrated by the regional anatomy in this study. Similar to the two acupoints on the human body, they are innervated by the peroneal nerve and saphenous nerve, respectively . This peAlthough the neural components are the same for the innervation of both LR3 and LR8, their segmental and regional distribution is markedly different between each other. For the spinal segments, the concentration of sensory and motor labeling associated with LR3 is obviously one segment lower than that of LR8. For the target regions, it is mainly exhibited on the distribution of the transganglionic axon terminals and motor neurons. The transganglionic axon terminals associated with LR3 locate more medially in the spinal dorsal horn than that of LR8, and the motor neurons associated with LR3 are situated more laterally on the spinal ventral horn than that of LR8. These segmental and regional distribution is accorded with the “somatotopic arrangement” in the CNS of rats, which has been well defined in the previous studies . Increasvia these central relay stations to the contralateral side of ventral posterolateral thalamic nucleus and the ipsilateral cerebellum through the spinothalamic, medial lemniscus, and spinocerebellar tracts, respectively (According to the distribution of the transganglionically labeled axons in the CNS, our results imply that the acupuncture signals from the LR3 and LR8 can be transported to their corresponding targets in the spinal dorsal horn, Clarke’s nucleus, and gracile nucleus, and forwarded ectively . ComparaSince acupoints are considered as the specific sites for acupuncture, how to select a proper acupoint becomes an important issue to be continuously concerned in the clinical application . GeneralFrom the viewpoint of neural pathways, present neuroanatomical findings from the regional anatomy and neural tract tracing suggest that although LR3 and LR8 are directly innervated by the peroneal and saphenous nerves, respectively, they may also involve in their successive parts of sciatic and femoral nerves, even in their corresponding spinal segments and target regions. Accordingly, the acupuncture signals aroused from LR3 and LR8 might be transported along these neural pathways to the CNS for the precise target treatment. Therefore, the results of this study provide valuable references for evaluating the therapeutic roles of LR3 and LR8.The primary focus of this study was the identification and characterization of the differential innervation between the LR3 and LR8 at the anatomic and cellular levels. Although our study provides the neuroanatomical pieces of evidence that the sensory and motor pathways may participate in the processes of signal transportation from the LR3 and LR8 to the CNS, their functional effects remain to be established experimentally.Due to the technical limitation, all the neural labeling with CTB and AF594/488-CTB along the sensory and motor pathways belongs to the first-order neural components associated with the LR3 and LR8. Their successive higher-order neural components remain unclear, which might be transsynaptically traced with neurotropic viruses in the future study . In addivia the sensory and motor pathways. These results suggest that when we select the proper acupoints for the acupuncture treatment, the neuroanatomical specificity of acupoints should also be taken together as the clinical demands, besides the consideration from the traditional theory of meridians.This study evidently showed the neuroanatomical characteristics of LR3 and LR8. Although both acupoints are attributed to the same meridian, considering their sensory and motor innervation, each of them establishes its own neural connection with corresponding spinal segments and target regions from the PNS to the CNS The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The animal study was reviewed and approved by the Ethics Committee of the Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences.The manuscript presented here was carried out in collaboration between all authors. DX, LZ, WZ, JL, YSu, YG, and YSh carried out the experiments. JW and JC analyzed the data. WB designed the study. DX and WB wrote the article. All authors read and approved the final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


In recent years, great progress has been made in 3D simulation modeling of instant network communication system, such as the application of virtual reality technology and 3D virtual animation online modeling technology. Facing the increasing demand of different industries, how to build an instant network communication system for 3D virtual animation has become a research hotspot. On this basis, the construction method of fast instant network communication system based on convolutional neural network and fusion morphological 3D simulation model is studied. This paper analyzes the research status of instant network communication system. The experiment optimizes and improves the shortcomings of the current research hotspot of virtual animation instant network communication system and takes the morphological 3D simulation model fusion as the core for in-depth optimization. Finally, the experimental results show that the fusion morphological 3D simulation model can reconstruct the standard 3D virtual animation model according to different needs and can quickly model the optimization strategy according to the local differences of different animations. The response accuracy of the network communication system reaches 97.7%. There are many problems in 3D virtual animation, such as low authenticity and slow modeling speed . How to The contributions of this paper are as follows. (1) We combine the deep convolutional neural network algorithm to propose a 3D virtual animation-oriented instant network communication system and a three-dimensional fast modeling model. (2) We study the method of 3D reconstruction of virtual animation, according to the local feature differences, feature tracking, and modeling speed between different 3D virtual animation models, and a 3D virtual animation-oriented 3D based on fusion form is constructed. (3) We build an instant network communication system for 3D virtual animation.The rest research is as follows. In recent years, David et al. have studied the application of 3D virtual animation fast modeling algorithm in the network communication system . FengyinAccording to the current common types of virtual animation and the standard range of animation size, the size of the 3D animation model to be established is obtained . In thisThe fast modeling algorithm of 3D virtual animation and the intelligent processing model of local data information based on the existing animation proposed in this paper adopt the multilayer fusion morphological 3D simulation model and structure. It is assumed that the first input value of the target, the hidden input value of the security feature, the output layer data of the security judgment, and the number of nodes needed for the fast modeling algorithm of 3D virtual animation are known. The data description of the 3D local model adopts the standard evaluation method.The instant network communication system of 3D virtual animation and the fast modeling method of 3D simulation are studied. The second part introduces the research status of instant network communication system technology and analyzes the possibility of innovation according to the existing research results. The fast modeling algorithm of 3D virtual animation based on the fusion morphological 3D simulation model is constructed, the optimization method of instant network communication system based on Gaussian random distribution is proposed, and the quality evaluation system of the instant network communication system is constructed. The experiment of the instant network communication system and three-dimensional simulation algorithm is designed. The experimental results are analyzed and the conclusion is drawn. Therefore, the innovation of this study is to propose a fast modeling method of 3D virtual animation based on morphological 3D simulation model fusion. Based on the rough recognition of local features of animation, this model makes full use of the perceptual information of animation to realize the dynamic and fast modeling of virtual animation. The similarity and consistency between the comparison bar  and the reference bar  is quantitatively described. It can make full use of the local differences of each virtual animation to realize the rapid modeling of information transmission of the instant messaging system in 3D simulation.The dynamic local tracking process of 3D virtual animation based on convolution neural networks is mainly divided into the basic model and the structural model. The basic model includes the sequential model and the parallel model, and the structural model includes the cyclic model . CombineAccording to the characteristics of collaborative simulation process, the local feature reconstruction process model can be divided into the basic model and the control model . Accordip1 and p3 are all serial structures, the design process p3 and p1 are conditional serial structures, and p is conditional probability. When it is less than or equal to p2, after the design process p3 is executed, the whole design process ends without iteration. In view of this situation, the 3D virtual animation reconstruction model of the design structure in the previous section is extended, and the 3D animation modeling is carried out through the multivalue fusion morphological 3D simulation model [T represents the calculation period of each animation modeling process node, the sequential coupling degree between Do process nodes, and the local facial coupling degree set between Dss process nodes. According to the definition, the calculation period of each local 3D simulation process node is t, and the number of motion process nodes is n. The calculation period of the whole local 3D simulation process is represented by the calculation period matrix Ad; then, Ad is a one-dimensional matrix, and its matrix value isThe design process on model . The mulon model . Let theThe corrected value ist, and the number of 3D simulation process nodes is n. The order coupling degree of the whole local 3D simulation process is expressed by the order coupling degree matrix Ab; then, Ab is a two-dimensional matrix, and the matrix value isThe order coupling degree of any two 3D local simulation process nodes is The modified matrix value isti, we can judge the order dependence between two process nodes. The judgment method is as follows: when i ≠ j, ai,j=1, ai,j=0, it means that the design process represented by row i and column j has order dependence. The order is as follows. If i < j, then the design process j is before the design process i. At this time, the matrix value isAccording to the value of i > j, then the design process i is before the design process j and the matrix value isIf i ≠ j, bi,j=1, bi,j=1, it means that the design process represented by row i and column j has a sequential coupling relationship, and the value of the judgment matrix isWhen i and j represent different sequences; the calculation cycle of the design process is different, so the sequence is determined according to the scheme of the shortest 3D virtual animation fast modeling time.The design process Cb isThe time of different collaborative local 3D simulation processes varies greatly; for example, a simple calculation process takes a few seconds. If the calculation period of local modeling process is a long period, the calculation period is divided by 5 seconds, and the value of the corresponding period matrix Cb|′ can be expressed asAfter coupling analysis and optimization, the value of matrix |T of the process is a dimensionless value. For simple multivalue fusion morphological feature model, diagonal is the calculation period; the first of other matrix values is sequential coupling degree, others are resource coupling degree, and zero value means uncoupled [In the process of matrix value optimization, if the calculation period of local simulation process belongs to the medium period, the calculation period is divided by 5 minutes and seconds at the same time. If the calculation period of the local 3D simulation process is a short period, the calculation period is divided by 5 ms at the same time. After normalization, the calculation period ncoupled .In this paper, we first simulate the common animation local features. According to the fusion morphological 3D simulation model and 3D virtual animation reconstruction algorithm proposed above, we simulate the distribution of common animation local features. The data processing flow of 3D simulation animation is shown in di, so as to achieve quantitative representation [di can be expressed asIn the process of 3D reconstruction of animation, different parts of the animation show different 3D simulation changes, and because of the local differences between each animation itself, there are also differences in the 3D simulation process between different parts and the simulation process of local features. Therefore, this study first collected a large number of animation common local data as training samples and then combined with access analysis software for local difference model reconstruction, to achieve the 3D reconstruction of different animation local posture, in order to help improve the effect and accuracy of the animation instant network communication system. In the process of local difference analysis, the value of difference degree can be calculated according to the feature factor entation . In the Considering the difference of different types of virtual animation in the feature value, the disturbance correction is made. At this time, the difference can be expressed asOn the contrary, in order to better analyze the local stability and standardization of animation, we also established a basic instant network communication system database in the experiment process to realize continuous updating and correction. The data processing flow is shown in di, that is, the first step is to decompose the known local 3D information of common animation and express and store it in the form of matrix. This constraint process can be expressed asOn the contrary, because this research is based on the convolution neural networks algorithm and is oriented to 3D virtual animation, so, in the process of simulation experiment, we need to add constraints according to the feature factor In order to improve the reliability of constraints, the constraint process can be expressed as follows:In this way, the local features of different animations can be dynamically changed and recognized. In this process, different fusion forms of 3D simulation models are introduced, and Gauss random matrix and Bernoulli matrix are used to store vectors to realize the recognition and storage of different types of 3D virtual animation communication information and gait matching. The simulation results of data are shown in As can be seen from Through the analysis of correlation function, the feature vectors of different gait vectors are solved, and the point product multiplication is carried out to realize the classification and differentiation of different gait vectors. Then, local feature detection and multidimensional reconstruction of animation are carried out, including video detection, image detection, and local feature factor-tracking detection. The simulation results of three groups of data are shown in From Considering the different local features of different virtual animations, it is necessary to add different animation reconstruction constraints to different 3D virtual animation modeling algorithms before formal experiments to avoid misjudgment in the process of virtual animation modeling. Therefore, the constraints of different virtual animation instant network communication systems will be realized from the following aspects. The preliminary results of the experiment are shown in The change trend of experimental data is similar, and the difference is not significant. This is because the fusion form of the augmented reality 3D simulation model is processed together with the known 3D virtual animation reconstruction model . The recThe experimental results are as follows. For different types of virtual animation samples, the effect of 3D reconstruction is different in the process of reconstruction for different 3D virtual animation. For example, for the virtual animation prototype in “Monkey King,” the 3D virtual animation rapid modeling algorithm will present two types of 3D virtual animation images. They are the conventional 3D virtual animation and the 3D virtual animation with the characteristics of Monkey King . Finally, the experimental results show that the fast modeling algorithm of the instant network communication system can realize the fast modeling of 3D virtual animation and the fast transmission of instant network communication and verify the effectiveness and feasibility of the fast modeling of 3D virtual animation. Therefore, the model can also provide a basis for the analysis of fast modeling algorithm of 3D virtual animation corresponding to 3D virtual animation in various environments. The experimental results are shown in It can be seen from The error analysis of the experimental results is shown in The function output image of the experimental results is shown in e = 0.012 in the quality evaluation of the 3D virtual animation fast modeling algorithm. From Tables According to the results in Tables In recent years, 3D modeling of 3D virtual animation has been well developed, but there are still many problems in the authenticity and rapidity of the algorithm of the network instant messaging system. How to use scientific intelligent model and convolution neural network algorithm to optimize has become a research hotspot. Based on this, this paper studies the instant network communication system of 3D virtual animation and the fast modeling model of 3D simulation. This paper first analyzes the research status of reality and signal transmission speed in the three-dimensional virtual animation instant network communication system, then optimizes and improves the shortcomings of the current research hotspot instant network communication system, and optimizes it combined with the fusion morphological three-dimensional simulation model. Finally, the experimental results show that the fusion of the morphological 3D simulation model can improve the authenticity and speed of 3D virtual animation rapid modeling, and the standard protocol of the instant network communication system can be formulated according to different 3D virtual animation elements, and the accuracy of 3D modeling can reach more than 97.7%. However, this paper only studies the real-time network communication system from the perspective of enhancing realism, without considering the potential impact of local differences on animation, so the network communication system of 3D virtual animation needs further research. Therefore, future work can be normalized research and processing from the local differences and influencing factors of 3D animation and combined with quantitative evaluation criteria for analysis.