MED12, HMGA2, and FH genes should be considered when developing or prescribing drugs. For example, synthetic inhibitors and vaccines based on matrix metalloproteinases MMP11 and MMP16 are expected to be effective only for the prevention of the occurrence of MED12-dependent nodules.Uterine leiomyomas are the most common benign tumors in women of childbearing age. They may lead to problems of conception or complications during the gestational period. The methods of treatment include surgical  and therapeutic treatment . Both approaches are efficient but incompatible with pregnancy planning. Therefore, there is a call for medical practice to develop therapeutical means of preventing leiomyoma onset in patients planning on becoming pregnant. Based on the analysis of GWAS data on the search for mononucleotide polymorphisms associated with the risk of leiomyoma, in meta-transcriptomic and meta-methylomic studies, target proteins have been proposed. Prospective therapeutic treatments of leiomyoma may be based on chemical compounds, humanized recombinant antibodies, vaccines based on markers of the uterine leiomyoma cells that are absent in the adult organism, or DNA and RNA preparations. Three different nosological forms of the disease associated with driver mutations in the  Uterine fibroids are most common in women aged 35–50 years (frequency of 30–35%), but there is a tendency for the first onset of the disease at an earlier age—25 years and even younger . In the According to data from various sources, among women of reproductive age, the incidence of the disease varies from 30 to 50%. The average age of detection of uterine fibroids is 32.8 ± 0.5 years, and indication for surgical treatment occurs after 4.4 ± 0.3 years . In primAmong women diagnosed with uterine leiomyoma, 15–30% have symptoms of varying severity, including pain syndrome, infertility, dysfunction of adjacent organs, abnormal uterine bleeding, anemia, and a number of other severe complications . DespiteIn gynecological hospitals in Russia, up to 50–70% of operations are performed specifically for symptomatic uterine leiomyoma. In addition, 40 to 60% of hysterectomies are performed specifically for this disease. In European countries, more than 300,000 surgical interventions related to uterine fibroids are performed annually , and in Fibroids are capable of rapid growth during pregnancy, which can lead to violations of the normal growth of the placenta and the development of defects in the fetus. Often, such problems might lead to the performance of urgent myomectomies in the early stages of pregnancy or cesarean section before the delivery date. Fibroids are able to grow rapidly in size during pregnancy, and in the case of subserous localization, tumors form adhesions with internal organs, which cause serious difficulties for uterine contractions during and after childbirth. In addition, there are cases of postpartum bleeding due to uterine atony .The development of postoperative relapses is a separate problem in the treatment of the disease, since the recurrence rate after myomectomy reaches 90%, while in almost every fourth case there is a need for repeated treatment until a new surgical intervention. The probability of recurrence depends on how thoroughly all the existing fibroids were removed; however, in the presence of multiple tumors, the risk of recurrence is higher, since often small fibroids are not visualized and, as a result, are not removed, continuing to grow in the postoperative period. According to the review by , the risHysterectomy is a radical way to prevent the recurrence of leiomyoma; however, it is not acceptable for women planning pregnancy. Moreover, a significant problem arising as a result of a myomectomy is the traumatization of the uterus, the occurrence of scars that lead to the disruption of its normal functioning during pregnancy, and the risk of ruptures during stretching and contraction before and during the course of childbirth. In addition, the growth rate of fibroids accelerates during pregnancy, which is usually explained by a high level of blood progesterone . Thus, tHMGA2 gene overexpression as a driver mutation in UL; (2) the driver mutations in the MED12 gene in UL; (3) the driver null-mutations in the FH gene in UL; (4) using the agonists and antagonists of steroid hormones as UL growth suppressors; (5) PI3K/Akt/mTOR signaling pathway in UL and its inhibitors as UL growth suppressors. The first three directions are important since ULs with different driver mutations differ substantially in the expression profile, and consequently, in the molecular targets of the candidate therapeutics. Direction 5 allows for analyzing the efficacy and side effects of commercially available and prospective medicines used for UL treatment. Directions 5 and particularly 6 allow for identifying novel molecular targets for UL drug and vaccine design that are prerequisites for fibroid formation and growth but absent in the normal myometrium. The key publications of each direction are summarized in We suggest specifying the following principal directions of uterine leiomyoma etiology studies and the development of approaches to the treatment of this condition: (1) Using the cited publications, the following sections are formed in this review: Currently, the molecular mechanism of the occurrence of the fibroids remains unclear . It is aMED12 gene (codons 130 and 131 and in frame deletions) encoding the regulatory subunit of RNA polymerase 2, and rearrangements leading to overexpression of the HMGA2 protein, a non-histone chromatin-binding protein involved in the regulation of the chromatic condensation and regulation of gene expression. These driver mutations account for more than 95% of cases of leiomyoma. In addition, biallelic inactivation of the fumarate hydratase gene (Krebs cycle enzyme) and rare chromosomal rearrangements, in particular, the loss of the Xq22.3 region, leading to the loss of COL4A5 - COL4A6 collagen genes, can act as a driver mutation [In 2017, the concept of a driver mutation leading to the appearance of a fibroid was established in the literature . It is nmutation .MED12-dependent and HMGA2-dependent fibroids differ significantly. A feature of MED12-dependent fibroids is a high proportion of fibroblasts , a high content of extracellular matrix (ECM), and a low degree of vascularization. The fibroids of this type have a relatively small size but show a tendency of multiplicity. Due to the high content of fibroblasts, the growth of MED12-dependent fibroids is stimulated not only by progesterone, but also by estrogens, which is not typical of other fibroid types.Conflicting opinions have been expressed in the literature regarding the possibility of two driver mutations co-existing in one cell. Most authors suggest that two mutations in one cell are incompatible. In works ,61,62,63HMGA2-dependent fibroids do not contain a clearly determined sensu stricto driver mutation, but their common and fundamental feature is the overexpression of the HMGA2 gene, and sometimes also of the HMGA1 and PLAG1 genes, which determine the overall expression profile and metabolism of the cell. The most common cause of HMGA2 overexpression may be the translocation of the 12q14.3 region containing the HMGA2 gene to chromosome 14 or to other parts of the genome, where the gene falls under the control of promoters of other genes (most often RAD51L1), or its 3′-terminal region is deleted, contributing to the destabilization of the transcript. The second common cause of HMGA2 overexpression is deletion of the long arm of chromosome 7 (q21.2q31.2), which contains the regulatory RNA genes miR-21, miR-23b, miR-29b, and miR-197 [HMGA2 expression, so their loss leads to overexpression of the regulated gene. More than half of the fibroids with HMGA2 overexpression do not have detected chromosomal rearrangements, and HMGA2 overexpression is usually suggested to be achieved there by changing the methylation status [HMGA2 overexpression significantly complicates the identification of such fibroids at the level of laboratory diagnostics; the most reliable way to solve this problem is to detect the HMGA2 protein using immunohistochemical staining or its transcript with RT-qPCR. The accuracy of diagnosis is facilitated by the absence or extremely low level of HMGA2 expression in normal myometrium, although there are reports of the possibility of moderate HMGA2 expression in MED12-dependent fibroids. miR-197  and the  miR-197 . These mn status . The preBiallelic inactivation of the fumarate hydratase gene is characteristic mainly of a rare type of leiomyoma with bizarre nuclei . AccordiMED12 gene nor overproducing HMGA2 are prone to so-called chromotripsis . Due to the small size of the rearranged fragments of genomic DNA, such rearrangements are difficult to detect by methods of classical cytogenetics.Other driver mutations occurring in the fibroids are so rare that their systematic study on the basis of statistically reliable data is practically difficult. Information about these mutations is present in the works by ,62. It wHistorically, the first methods of therapy for leiomyomas were the use of hormone antagonists or antitumor cytostatics acting at the level of the PI3K (phosphatidylinosylkinase 3)/Akt/mTOR signaling pathway (target of the antibiotic rapamycin inhibiting autophagy) . This isHMGA2 [A consistent increasing production of pACT and GF2BP2 was observed in the cells of the fibroids with an overexpression of HMGA2 . The speThe suppression of pACT expression with miR-182 micro-RNA or with a chemical inhibitor MK-2206 was reported to lead to an increase in the production of proteins p16 (Ink4a), p53, and p21, which ultimately causes the phenomenon of premature senescence in leiomyoma cells . HoweverCompound T-5224, an inhibitor of transcription factor AP1 (activating protein 1) involved in the activation of ECM synthesis and the formation of fibrosis foci, was described by  reportedThe use of gonadotropin-releasing hormone (GnRH) agonist trials (leuprolide acetate) and GnRH antagonists (cetrorelix acetate) as a candidate therapy for leiomyomas is described by . It was The use of selective progesterone receptor modulators , antiprogestin (mifepristone—RU486) is described in . The treThe effects of goserelin, an agonist of luteinizing hormone-releasing hormone, on serum leptin levels and body composition in women with uterine leiomyoma were reported by . FifteenClostridium histolyticum collagenase (CCH), which selectively hydrolyzes collagen types I and III and changes the stiffness of the ECM, in combination with verteporfin, an inhibitor of the YAP factor (Hippo/YAP pathway) and the antifibrosis drug nintedanib were reported in [3 into fibroids after 60 days resulted in a 46% decrease in stiffness compared to the control. The level of PCNA, a marker of cell proliferation of nuclear antigen of proliferative cells, was reduced 60 days after the injection of high doses of CCH. The contents of key intracellular growth signaling factors Hippo and the phosphorylated form of YAP (p-YAP), as a result of the use of CCH, were increased, which contributed to maintaining a high rate of fibroid growth. The inhibition of YAP by verteporfin reduced cell proliferation, gene expression, and the proteins of key factors contributing to fibrosis and mechanotransduction in fibrous cells. The antifibrotic drug nintedanib additionally reduced the activity of YAP and showed an antifibrotic effect.The results of clinical trials for the treatment of uterine leiomyoma with orted in . The intTrials of the plant-derived polyphenolic compound resveratrol for the treatment of leiomyoma are described in . The autA method of therapy for leiomyomata with a driver mutation in the fumarate hydratase gene was proposed in . They clMED12 gene, HGMA2 overexpression, and biallelic inactivation of the fumarate hydratase gene are unlikely to completely coincide. Therefore, now there is no possibility of predicting the risk of each type of leiomyoma based on the analysis of the patient’s genotype.To date, a number of works have been carried out worldwide that are aimed at identifying factors of hereditary predisposition to the occurrence of leiomyomas ,71,72,73There are also a number of papers in the literature focused on the molecular profiling of leiomyoma at the level of transcriptome and DNA-methylome analysis ,61,62,63MED12 and those with an overexpression of HMGA2. This conclusion is fully supported by the work of [The work by  confirms work of . Therefo work of , prolact work of  also fou work of  demonstrMED12-dependent and HMGA2-dependent types at the level, significantly exceeding those in the normal myometrium. They should be considered promising targets for therapy. However, for the experimental verification of the effectiveness of candidate therapeutic agents, it is advisable to narrow down the fairly wide range of these targets, giving preference to two categories of proteins:Secreted proteins that do not remain on the surface of the cells producing them;Proteins present exclusively in leiomyoma cells and absent in any cells of normal tissues.More than 40 genes are listed in ,61,62, t3.PCP4—Purkinje cell protein 4, expressed in embryogenesis; CHRDL2—growth factor, a chordin-like antagonist of BMP; RPE65—retinol transporter from the retinal epithelium; MMP11—matrix metalloproteinase 11 (or stromelysin 3), expressed in embryogenesis, causes metastasis of breast tumors; MFAP2—microfibrillar-associated protein 2, affects the motility of cancer cells in gastric cancer, regulates the expression of a5ß1 integrin via the ERK1/2 pathway.From the work by : PCP4—Pu4.From the work by :HMGA2-dependent fibroid types: HMGA2, GRPR—gastrin precursor; PLAG1—proto-oncogene, the main participant of HMGA2-dependent signaling; PAPPA2—pappalysin, a protease that destroys IGFBP5, a marker of osteoblasts; MB21D2—nucleotidyltransferase, an anti-virus protection gene induced by interferon;For MED12-dependent fibroid types: ADAM12—membrane-bound metalloproteinase responsible for shedding—cleavage of extracellular receptor domains and proteoglycans; MMP11—matrix metalloproteinase 11 (or stromelysin 3), expressed in embryogenesis, causes metastasis of breast tumors; MMP16—matrix metalloproteinase 16, has a transmembrane domain, expressed in embryogenesis; RAD51B—protein of the DNA reparation system, similar to RecA, with ATPase activity, not expressed in normal tissues; PCP4 is a protein of 4 Purkinje cells, expressed in embryogenesis; RUNDC1 is a DNA-binding protein with a RUN-type domain at the C-terminus; THSD4 (Adamtsl6-β) matrix metalloproteinase with a thrombospondin domain associated with the cell surface; participates in the formation of ECM microfibrils, ensuring elasticity and release of deposited growth factors. Adamtsl6-β expression increases during the formation of the periodontal ligament, which consists of a fibrillin-1 microfibril.For Such an approach to the selection of targets is dictated by the need for minimizing the side toxicity of candidate drugs, which can be achieved only if normal organs and tissues remain unaffected. In the first case (the secreted molecules), the proteins themselves and not the producer cells are targeted, and the therapy leads only to temporarily reducing their concentration in the extracellular space, while the producer cells remain alive. Therefore, requirements for the absence of the targets in normal cells may be not absolute. As for proteins present strictly in leiomyoma cells, they are found mainly among embryonic proteins that are absent in adult cells, but they often act as cancer markers. After sorting the potential targets according to the proposed principle, the following list of genes (proteins) can be presented:IL17B mRNA in leiomyoma. IL17B is a cytokine derived from T-lymphocyte and involved in the initiation and maintenance of inflammation, fibrosis, and keratinization. An advantage of IL17B as a target for leiomyoma therapy is the availability of recombinant humanized antibodies approved for the treatment of psoriasis and psoriatic arthritis [An Interesting finding by  is the drthritis . These dRPE65), which was identified in the same work as a marker of differential expression in leiomyoma, may also be considered a target available for therapy with pre-existing drugs. It can be assumed that the superexpression of RPE65 in leiomyoma cells makes them sensitive to the synthetic retinol analogs used in pharmaceuticals for the treatment of acne: adapalene, tretinoin, isotretinoin, and tazarotene [The protein retinol transporter from the retinal epithelium (zarotene . These p(1)IL17—for all leiomyoma types;The immunobiological drugs secukinumab, ixekizumab, brodalumab, and netakimab, a recombinant humanized antibody against (2)Synthetic analogues of retinoids: adapalene, tretinoin, isotretinoin, and tazarotene—potential blockers of the RPE65 receptor—for all leiomyoma types;(3)PARP1, also known as NAD+-ADP-ribosyltransferase 1 and PARP2)—for leiomyoma with biallelic inactivation of the fumarate hydratase gene as a driver mutation;Olaparib and niraparib—synthetic inhibitors of poly-ADP-ribosopolymerase ((4)Resveratrol—for all leiomyoma types.The development of new therapeutic agents should be started by testing those pharmaceutical substances that are already available for use and have passed preclinical clinical trials to the maximum extent . Among tMMP11, MMP16, and THSD4 (Adamtsl6-β) should be considered a quick and easily accessible way to suppress their activity in vivo. The review by [The development of inhibitors based on amino acid hydroxamates, carboxylates, thiols, phosphonates and sulfonamides, flavonoids, or 15oxycycline derivatives against matrix metalloproteinases ADAM12, eview by  containseview by , clinicaMED12, overproduction of HMGA2, and biallelic inactivation of FH) were not used for the development of specific medicines directed against specific targets of the different nosological forms of uterine leiomyomata except for relatively rare FH-dependent cases.Since 1987, the development of medicines for the treatment of uterine leiomyoma and basic studies of the molecular mechanisms of the initiation of this disease were carried out in parallel without proper mutual coordination. For instance, goserelin acetate was patented in 1976 and approved by the FDA in 1983 . MifepriMED12 gene should be considered a separate relevant object for the development of therapeutics. This follows from the greatest prevalence of this type of leiomyoma, as well as from the large number of target proteins in it, which are completely absent in normal tissues of the adult human body but necessary for the survival and growth of the tumor. This category should include, first of all, free and membrane-bound metalloproteinases: ADAM12, MMP11, MMP16, and THSD4 (Adamtsl6-β). The development of specific inhibitors to them is a routine procedure, delivery to the target is facilitated by its extracellular location, and the absence of targets in normal tissues will ensure the absence of undesirable side effects. It also seems promising to create inhibitors of the potential-dependent channels KCNAB3 (potassium-specific) and CACNA1C , as well as hyaluronidases that stimulate cell migration (a product of the KIAA1199 gene).Analysis of the data by  suggestsMMP11 seems to be the most promising target for the development of target therapy of leiomyoma, since it is not expressed in any of the normal tissues of the body; however, a sustainable therapeutic effect of leiomyomata is expected only if the MMP11 inhibitor is combined with blockers of some other targets. Trials of these inhibitors should be carried out mainly on patients with MED12-dependent leiomyomas. It can be assumed that these compounds will have low side toxicity and will be suitable for patients at all stages of pregnancy. Developing an inhibitor of pappalizin (PAPPA2), a protease that destroys IGFBP5 and IGF1 mRNA binding protein, a marker of osteoblasts, is also technically accessible. In this case, the trials should be carried out on a group of patients with HMGA2-dependent leiomyoma.CACNA1C, is also a technologically easy task. Trials of these inhibitors should be carried out on patients with MED12-dependent leiomyoma.Creating inhibitors of membrane receptors, potential-dependent channels KCNAB3 and MMP11 and MMP16 metalloproteases from the body also has good prospects. Their trials should be carried out on groups of patients with MED12-dependent leiomyomata. In addition, these drugs can be effectively used for the treatment of a number of malignant solid tumors, in particular, to prevent the metastasis of colon carcinoma. The prospects of this approach are confirmed by the data of [MMP11 to protect mice from colon carcinoma. Immunization not only protected the animals from tumor progression but also did not show undesirable side effects.A modern and effective way to remove unwanted proteins from the body (blocking their activity) is the development of immunobiological drugs based on humanized recombinant antibodies. An experimental approach allowing for the selection of targets for the development of such antibodies is presented in  data of , which pSatb2, Nrg1, PCP4, CHRDL2, and MFAP2—to combat all leiomyoma types; HMGA2, GRPR, PLAG1, PAPPA2, and MB21D2—to combat HMGA2-dependent leiomyoma; ADAM12, MMP11, MMP16, RAD51B, THSD4 (Adamtsl6-β), and RUNDC1—to combat the MED12-dependent leiomyoma types. Vaccines can be either protein or genetic (based on DNA or RNA). Vaccines of the first type require longer development and testing but have a significantly lower manufacturing cost per dose, which makes them more accessible to patients. The great advantage of vaccination is safety for patients compared to chemical agents, which allows them to be used during pregnancy. The advantage of vaccines is also the possibility of the immunization of patients planning pregnancy with a high individual risk of leiomyoma onset, whereas the effect of immunization will be manifested during pregnancy, when the use of any medications is limited. An indirect argument confirming the effectiveness of vaccination as a means to combat the occurrence of leiomyomata is the fact that the risk of developing leiomyoma decreases significantly with each full-term pregnancy, whereas aborted pregnancies, on the contrary, increase the risk of the disease [Satb2, Nrg1, PCP4, CHRDL2, MFAP2, HMGA2, PLAG1, PAPPA2, MB21D2, ADAM12, MMP11, and MMP16. The immunogenicity of the products of these proteins is probably not high by itself or due to the presence of the placental barrier, but due to repeated boosting during multiple pregnancies, the production of antibodies specifically suppressing the growth of the fibroids can gradually reach a significant level.In addition, an economically affordable way to protect patients against leiomyoma onset is the creation of preventive and protective vaccines based on proteins with high specificity for leiomyoma cells. An experimental approach allowing for the selection of targets for the development of vaccines coincides with one applicable for developing immunobiological preparations  . A large disease . The reaIt is noteworthy that the effectiveness of the use of preventive vaccines to prevent the development of leiomyomas will depend on the development of diagnostic tools that allow for determining the individual risk of developing a certain type of leiomyoma in a patient. This task is currently still waiting for its resolution.HMGA2-dependent leiomyomas, the delivery of miR-21, miR-23b, miR-29b, and miR-197 micro-RNAs described in [HMGA2. A total of 45 micro-RNAs with significantly increased or decreased content in the fibroids compared to the corresponding myometrium were identified.Finally, a promising area of leiomyoma therapy is the delivery of RNA and DNA constructs. For example, in the case of ribed in , which sMED12- and HMGA2-dependent leiomyomata, participating in the regulation of the expression of the chromatin demethylation factor TET3.A good effect can be expected from the delivery of miRNA to induce the specific degradation of the long non-coding RNA H19 described in . This RNCEACAM1 (carcinoembryonic antigen-cell adhesion molecule 1), described by [KLF11 genes [ribed by  and KLF111 genes ,78 shoulHMGA2-dependent leiomyomata, GPR20 and IL11RA can be specific receptors for solving this problem, since according to [MED12-dependent leiomyomas, this function could be carried out by the membrane proteins POPDC2, PLP1 , THSD4 (Adamtsl6-β-transmembrane protein 4 with thrombospondine domain).The production of RNA and DNA structures is a routine technological task, well-developed during the fight against the pandemic of the SARS-CoV-2 virus. However, for the targeted delivery of such molecules to the uterine leiomyoma cells, it is advisable to use functionalized vectors with specific affinity to the surface markers of these. In the case of rding to , their hWe anticipate that our study will promote the implementation of the data obtained by the modern high through-put molecular and bioinformatic methods for the development of efficient specific methods for mitigating the negative impact of leiomyomata on the life quality of the population.Satb2, Pcp4, Wif1, MMP11, MMP16, and Adam12 are the most unique molecular markers expressed in all types of fibroids , but silent in normal myometrium. These proteins should be investigated as candidate vaccine substances for the prevention of relapses of leiomyomatosis in patients with hereditary predisposition to this disease, particularly in those who are planning pregnancy. Additional candidate markers for the development of preventive and curative vaccines against leiomyomatosis can be identified as a result of immunological screening of blood sera from women who have given birth many times. Resveratrol and synthetic retinol analogs adapalene, tretinoin, isotretinoin, and tazarotene should be investigated as promising therapeutic agents that allow for slowing the growth of fibroids, including in patients with pregnancy.The results of the metatranscriptomic and metamethylomic analyses suggest that the proteins


The high adiabatic temperature rise and low heat dissipation rate of mass concrete will promote rapid hydration of the cementitious material and rapid consumption of water from the concrete pores, which may significantly accelerate the development of concrete autogenous shrinkage. In this study, the effect of the water-binder ratio on the autogenous shrinkage of C50 concrete mixed with MgO expansion agent (MEA) was explained with respect to mechanical properties, pore structure, degree of hydration, and micromorphology of the concrete based on a variable temperature curing chamber. The results show that the high temperature rise within the mass concrete accelerates the development of early (14 d) autogenous shrinkage of the concrete, and that the smaller the water-binder ratio, the greater the autogenous shrinkage of the concrete. With the addition of 8 wt% MEA, the autogenous shrinkage of concrete can be effectively compensated. The larger the water-binder ratio, the higher the degree of MgO hydration, and in terms of the compensation effect of autogenous shrinkage, the best performance is achieved at a water-binder ratio of 0.36. This study provides a data reference for the determination of the water-binder ratio in similar projects with MEA. High-strength concrete is being used more widely due to the increasing demands of modern construction ,2. CompaA great deal of research has been carried out and solutions have been proposed, both nationally and internationally, on how to reduce the autogenous shrinkage of concrete ,12,13. TUnder normal temperature conditions, the use of MEA to compensate for autogenous shrinkage of concrete has been well documented ,25. HoweThis study is based on a large volume straight wall concrete project, using the same concrete mix ratio as that of the straight wall on site (8 wt% of MEA). Through a simulation of the site temperature history (high temperature peak and low cooling rate), the effect of the water-binder ratio on the autogenous shrinkage of concrete with MEA under the variable working temperature conditions was investigated, in order to provide a basis for the selection of the water-binder ratio of concrete for engineering structures. In this research, Ordinary Portland cement (OPC) was provided by Jiangnan Onoda Cement Co., Ltd. . Fly Ash (FA) was supplied from Nanjing, and blast furnace slag (BFS) was provided by Mabel Ltd. , which were used as the supplementary cementitious materials. The MEA was supplied by Nanjing Subote New Material Co., Ltd. , of which the reactivity value was 200 s. Its reactivity was estimated by the citric acid method in accordance with Chinese standard CBMF-2017 . The cheFour concrete mixtures (numbered from C1 to C4) containing 8 wt% MEA are shown in A variable temperature curing chamber was used to simulate the history of changes in the central temperature of straight walls in engineering. According to Chinese standard GB/T 50081-2019 , the conConsidering that the concrete of the straight walls in the project was in a sealed state before the early demolding, which would have exacerbated the autogenous drying shrinkage, the autogenous deformation test was performed. The test for autogenous shrinkage of concrete is carried out using the buried strain gauge method with reference to SL/L 352-2020 . After t2 atmosphere, heated from room temperature to 950 °C.At 1 d, 3 d, and 7 d, the mixtures of concrete without sand were analyzed to calculate the hydration of magnesium oxide. After being cut into pieces, the samples were soaked in the anhydrous ethanol for a week for the purpose of terminating their hydration. Then, the samples were placed in a vacuum drying oven at 60 °C until the mass was stable. After, those dried samples were ground into powders of no more than 0.080 μm. The powder samples were tested using NETZSCH STA 449F1 thermogravimetry under a NThe pore structure of concrete specimens was tested using a mercury intrusion porosimeter (MIP). According to the sample preparation requirements, the samples maintained in the variable temperature oven were taken out. Concrete specimens were cut into 3~5 mm pieces and soaked in anhydrous ethanol, which was aimed at terminating the hydration of samples. Afterward, the samples were placed in an oven at the temperature of 60 °C until the mass was stable.2. Based on Mo [2 and the hydration degree of MgO can be calculated by Equations (1) and (2):ed on Mo , the qua2 and the hydration degree of MgO in the samples. It is clear that there is a rapid increase in the hydration degree of MgO during the first 3 d. The water-binder ratios  are 53.1%, 52.3%, 55.2%, and 67.4%, respectively, which are highly dependent on the curing temperature at which the specimens are maintained, once again confirming the temperature sensitivity of the MEA and that high temperatures accelerate the hydration of MgO. The hydration degrees of MgO in concrete with different water-binder ratios are relatively close at 3 d, which is probably attributable to the fact that there is sufficient water inside the concrete in the early stages and MgO competes with cement for water. When at 7 d, there is a notable difference between different water-binder ratios. The hydration degrees of MgO in the concrete with the water-binder ratios of 0.40, 0.36, and 0.40 are 72.6%, 66.9%, and 62.8%, respectively, whereas the hydration degree is only 58.9% when the water-binder ratio is 0.28. Combined with the changes of humidity in +, which allows more Mg(OH)2 products to grow inside the pores, thus changing the pore size distribution inside the pastes and refining the pore size.2 crystals are mainly in the form of flakes of different sizes, which grow near the MgO particles. Ca2+ has an inhibitory effect on the diffusion of Mg2+ and promotes the growth of Mg(OH)2; therefore, Mg(OH)2 crystals generally grow near Ca(OH)2 crystals. The lamellar Mg (OH)2 crystals in 2. The thermal analysis shows that there is little difference in the degree of hydration of MgO in concrete with different water-binder ratios at 3 d. However, at 7 d, the hydration degree of MgO in concrete with a water-binder ratio of 0.40 is 72.6% compared to 58.9% with a water-binder ratio of 0.28. Sufficient water is supplied to MgO to produce Mg (OH)2, which counteracts the pore pressure and is the key to compensating the autogenous shrinkage of concrete. Meanwhile, Ca2+ has an inhibitory effect on the diffusion of Mg2+, which promotes the nucleation and growth of Mg (OH)2. Thus, Mg (OH)2 crystals mostly grow overlappingly in pores not far from the MgO particles. Moreover, the higher the water-binder ratio, the denser the hydration product of MgO. The early arrival of the hydration period of MgO allows the crystalline growth pressure of the Mg (OH)2 crystals to resist the early shrinkage deformation of the concrete. However, the autogenous deformation of mass concrete in practical projects is more complex, as the restraint of reinforcement, the large temperature difference between day and night, and the method of construction all affect the autogenous shrinkage of concrete. Increasing or changing the water-binder ratio alone will not completely overcome the problem of autogenous deformation of concrete and needs to be carried out in cooperation with other effective measures according to the actual situation of the project at the time. This study was carried out based on a large volume of straight wall concrete and is intended to provide a reference for similar projects with MEA.Bulk high-strength concrete generally has a high adiabatic temperature rise due to the great amount of cementitious material and the slow dissipation of heat from the concrete. In large volume straight wall concrete projects, temperatures in the center of the concrete can reach over 70 °C (and even 80 °C) and do not drop to room temperature until 14 d. The high temperature changes within the bulk straight wall concrete not only accelerate the development of autogenous concrete deformation but also promote the hydration of MgO. Although the addition of MEA mitigates the autogenous shrinkage of the concrete, the effect of different water-binder ratios on the deformation of concrete under variable temperature conditions varies slightly. Compared to concrete with a water-binder ratio of 0.28, an increased water-binder ratio not only reduces the autogenous shrinkage of the concrete itself but also contributes more to the hydration of MgO and the production of more Mg (OH)The internal temperature variations of C50 bulk straight wall concrete structures in practical projects were simulated in the laboratory. On this basis, the influence of different water-binder ratios on the compressive strength, autogenous shrinkage, pore size, and micromorphology of concrete with MEA was studied. The key conclusions are the following:The hydration process of MgO is accelerated under variable temperature curing. The addition of 8 wt% MEA reduces the compressive strength of the concrete, but the effect is minimal (no more than 5 MPa). In this experiment, the compressive strength of the concrete at 7 d can reach 85% or more at 28 d. High temperature accelerates the early self-shrinkage of concrete, especially within 7 d. Increasing the water-binder ratio can reduce the autogenous shrinkage of concrete. When the water-binder ratio is 0.40, the concrete deformation is slightly expanded. Compared to the reference group without MEA, the best effect of compensating for autogenous shrinkage is achieved by adding 8% MEA with a water-binder ratio of 0.36. Therefore, in actual projects, the water-binder ratio can be relaxed to 0.36 or even 0.40 under the premise of ensuring strength.2 crystals, appear as lamellar in the cement system, growing mainly in the pores not far from the MgO particles and overlapping each other. As the water-binder ratio increases, the MgO hydrates more and the Mg(OH)2 crystals become denser, which fills the pores better and counteracts the autogenous shrinkage of the concrete.At high temperatures, the hydration product of MgO, Mg(OH)


In the verification tests, the root mean square error (RMSE) of the measured value of the sensor was 0.02 m3/m3, the mean bias error (MBE) was ±0.009 m3/m3, and the maximum error was ±0.039 m3/m3. According to the results, the sensor, which features a wide detection range and good accuracy, is well suited for the portable measurement of soil profile moisture.Soil profile moisture is a crucial parameter of agricultural irrigation. To meet the demand of soil profile moisture, simple fast-sensing, and low-cost in situ detection, a portable pull-out soil profile moisture sensor was designed based on the principle of high-frequency capacitance. The sensor consists of a moisture-sensing probe and a data processing unit. The probe converts soil moisture into a frequency signal using an electromagnetic field. The data processing unit was designed for signal detection and transmitting moisture content data to a smartphone app. The data processing unit and the probe are connected by a tie rod with adjustable length, which can be moved up and down to measure the moisture content of different soil layers. According to indoor tests, the maximum detection height for the sensor was 130 mm, the maximum detection radius was 96 mm, and the degree of fitting (R The root systems of crops are distributed at different depths of the soil, and the soil water content differs at different depths. Therefore, the vertical distribution characteristics of soil water content seriously affect water absorption, growth, and development, and even the final yield of crops ,2. To reCurrently, the mainstream methods for measuring soil moisture content include the oven-drying method, dielectric method, neutronic method, tensiometry method, and remote sensing method ,9,10,11.The soil profile moisture sensor is isolated from the soil through a PVC pipe on the outer wall, and the noncontact measurement of the soil profile moisture is realized by using a single mobile probe or fixed multiple probes. Yuan et al.  designedIn this study, a low-cost, single-probe pull-out soil profile moisture sensor was designed following the principle of the capacitive dielectric method, for which the length of the tie rod can be flexibly configured to move it up and down a single-ring probe to measure the moisture of different soil layers, and a smartphone app is used to read the moisture content and other moisture information for the soil in real time. Compared with current moisture sensors, such as those based on time domain reflectometry (TDR) and frequency domain reflectometry (FDR), it does not require high-precision measurement circuitry and a complex secondary calibration process ,30,31,32Soil is a mixture of dielectrics, and the dielectric constant of water is higher than the dielectric constant of other solid particles by a quantity product ;C—Equivalent capacitance consists of probe and soil (F);S—Area of parallel plates (m2);d—Space between two parallel plates (m).out of the output is measured to calculate the soil moisture content –(6)). By fitting the relationship between this frequency and soil moisture, the soil moisture content under any condition can be inverted. The LC high-frequency resonant capacitor method can not only reduce the dielectric loss (shown in Equation (7)) but also ensure consistency between the soil moisture measurement results of different soil types because the polarization intensity of the soil medium is a function of frequency The sensor tie rod is designed as a segmental type, which is convenient to freely increase or decrease the number of segments according to the soil depth measured according to needs.(2)The positioning card ring is designed with a specially shaped card slot that matches the segment (this sensor is diamond-shaped), and the automatic positioning of the probe can be realized by simply raising the height of the tie rod to the corresponding soil depth when measuring, and then rotating a certain angle for misalignment.(3)They draw on mineral water bottles and design the protective shell of the data processing unit into a large and small middle at both ends and a profile structure that is easy to grasp.This set of devices has three advantages: The sensor is more suitable for portable measurements in multiple plots  , and theThe sensor is placed in the inner circle PVC pipe during the test, and the soil sample is filled in the outer circle PVC pipe. The test barrel shown in  is speci(1)Maximum Detection Range TestThe following tests were designed to obtain the sensor probe’s maximum detection diameter and maximum detection height. Among them, the test scheme of the maximum detection diameter was as follows: first, add dehydrated soil samples to 7 test barrels with different outer diameters Soil Volumetric Moisture Content CalibrationAs shown in (3)Laboratory Verification Test of Measurement Accuracy3/m3]. Put the soil samples into a large-enough transformation test barrel, and then store in a sealed barrel for 48 h at room temperature and measure the soil samples using self-developed moisture sensors. The actual volumetric moisture content of soil samples was measured using the oven-drying method and commercial moisture instrument WET-2. Using the value as a reference, and the root mean square error, the mean deviation error and maximum error of the laboratory measurement results of the self-developed sensor were calculated to verify the laboratory measurement accuracy of the sensor.Manually configure 35 soil samples with unknown moisture content with an obvious gradient; the moisture content interval was [0, 0.50 m(4)Field Verification Test of Measurement AccuracyTwo plots with obvious moisture gradients were selected outdoors, four test points were selected each, a hole with a diameter of 50 mm and a depth of about 400 mm was dug at each test point using a soil extractor, the bottom-sealed PVC pipe was inserted into each hole, and the top of the PVC pipe was sealed with a lid. After standing for 48 h, the self-developed sensor was inserted into the PVC pipe. The moisture content of different soil layers  was measured by pulling the tie rod up and down. A total of 24 soil samples of unknown moisture content were tested. As a control, a soil profile was dug near the test point (as shown in (1)Drying method for measuring soil volumetric moisture contentV) soil sample, and put it into an aluminum box; weigh out the total weight (M). Second, put the aluminum box with the lid removed and put it into the drying box for drying, take it out, and weigh it again as m. Finally, wρ represents the density of water in the soil (1 at room temperature), ∆M represents the reduced mass of the soil after drying, and the volumetric moisture content of the soil (Ɵ) is calculated according to Equation (9).The drying method is the main means of measuring the soil volume moisture content value and mass water content. The detailed procedure of the drying method is presented as follows: First, use a ring knife to take a fixed volume ((2)Sensor detection radiusL is the maximum detection radius or height of the sensor, x and y are the horizontal and vertical coordinates of the response curve, respectively.(3)Sensor measurement accuracy evaluation indexAccording to the relevant literature and product specifications of moisture sensors ,37, the (1)Root mean square error (RMSE)N is the sample size, M is the measured (soil sampling) value, P is the predicted (sensor measurement) value, and M is the average measured value.RMSE reflects the difference between the measured value of the moisture sensor and the actual moisture content value of the soil. RMSE is calculated according to Equation (13), where (2)Mean bias error (MBE)N is the sample size, M is the measured (soil sampling) value, P is the predicted value (sensor measurement), and M is the average measured value.The mean bias error (MBE) is the sum of the deviations of the individual measurements of the moisture sensor from the actual moisture content value, divided by the number of measurements, representing the difference of any value in the moisture content measurement of the soil sample. MBE is calculated according to Equation (14), where (3)Maximum errorM is the measured (soil sampling) value and P is the predicted (sensor measurement) value.The maximum error of the sensor refers to the maximum fluctuation between the measured value of the sensor and the actual value, which is used to reflect the worst-case scenario of the measurement, and the calculation is carried out according to Equation (15), where 2 is 0.9928; the curve equation of output frequency with soil sample height is 2 is 0.9949; according to Equations (11) and (12), the maximum detection radius of the sensor is 96 mm, and the maximum detection height is 130 mm.According to the test method of the maximum detection range in 2 is 0.9719.The moisture measurement model of the self-developed sensor shown in 3/m3, the average deviation error is ±0.033 m3/m3, and the maximum error is ±0.031 m3/m3. Taking the WET-2 measurement as a reference, the root mean square error of the indoor operating measurement of the sensor is 0.018 m3/m3, the average deviation error is ±0.050 m3/m3, and the maximum error is ±0.033 m3/m3.According to Equations (13)–(15), the root mean square error (RMSE), mean bias error (MBE), and maximum error of the laboratory measurement results of the self-developed moisture sensor are calculated with reference to the oven-drying method. The measurement value of the commercial instrument WET-2 and the results are shown in 2 of the curve obtained by linear fitting of the two is 0.9839 and 0.9802, respectively. The comparison results between the self-developed moisture sensor and the drying method and WET-2 are minimal. The WET-2 measurement and the drying method measurement are linearly fitting, and the R2 of the curve is 0.9949, which is extremely correlated. The absolute error distribution between the measured value and the actual value of the sensor is characterized by being large at both ends and small in the middle.As shown in 3/m3, the mean bias error (MBE) is ±0.009 m3/m3, and the maximum error is ±0.039 m3/m3 , mean bias error (MBE), and maximum error of the indoor measurement results of the self-developed moisture sensor are calculated according to Equations (13)–(15). The results show that the root mean square error (RMSE) of the self-developed sensor measurement results is 0.020 mshown in .2 between the two is 0.9066. The absolute error distribution between the sensor measurement value and the actual value shows the characteristics of the large right and small left ends.As shown in The development of portable and practical soil profile moisture sensors is a necessary means to improve the efficiency of soil moisture measurement and reduce the cost of soil moisture measurement, especially the application of soil moisture measurement in the multiple plots ,39,40; tIn the laboratory test, the absolute error distribution between the measured value of the sensor and the actual value shows the characteristics of being large at both ends and small in the middle. The analysis shows that the Jiangsu brown loam used for a laboratory test is a sample with different water content artificially configured after grinding, which is affected by the physical and chemical properties of the loam ,42. As a2 of the predicted value and the true value of the sensor in the field environment are lower than those in the indoor environment. The analysis shows that, in the field test environment, a soil extractor needs to be used to dig a hole, the embedded PVC pipe often has a certain air gap with the surrounding soil, and the electromagnetic wave intensity is attenuated exponentially in the air [The linear fitting curve R the air . If the  the air ,45.In this study, a portable pull-out soil profile moisture sensor was developed based on the high-frequency capacitance method. This work was mainly innovative in terms of the design of the sensor probe structure, the design of the data processing unit software and hardware, the shell and pull-out structure design, and the sensor performance testing and data processing.(1)The design of the sensor probe adopted an upper and lower ring electrode layout, which, together with the soil, forms a three-point LC resonant circuit equivalent capacitance. The MC12149 resonant circuit oscillates at 200 MHz, which reduces the influence of loamy-type soil on sensor measurements. The resonant frequency is divided twice by the frequency dividing/conditioning circuit, and finally, a low-frequency square wave signal that is easy to measure is obtained.(2)The data processing unit uses the Atmega328P microcomputer to measure the output frequency of the moisture sensor probe and communicates with the display terminal through the ESP8266 module. The lower computer uses the Arduino language for embedded system development based on the Arduino IDE environment, the upper computer uses the Java language for mobile app development based on the Eclipse environment, and the upper and lower computers use the Wi-Fi protocol for communication.(3)Using Solidworks to design segmental tie rods and gourd-shaped housings, the shell of the data processing unit was designed as a gourd-shaped package for easy grip, and the segmental tie rod and the card slot were misaligned to realize the hover measurement of the moisture-sensing probe. The tie rod and the card slot were used in order to enable the moisture-sensing probe to move up and down and allow the user to flexibly configure the segmented tie rod with different lengths according to the depth of the soil to be measured.(4)2 is 0.9719. The results of the laboratory and field accuracy verification test show that the root mean square error of the sensor was 0.020 m3/m3, the average deviation was ±0.009 m3/m3, and the maximum error was ±0.039 m3/m3.The data processing results for the laboratory soil sample testing of the sensor show that the maximum detection height of the sensor is 130 mm and the maximum detection radius is 96 mm. The equation of the moisture measurement model calibrated to obtain the sensor is (5)3/m3; MBE = ±0.02 m3/m3), and upon comparing the measurement accuracy of the developed sensor with that of the commercial sensor WET-2 (maximum error = ±0.03 m3/m3) using a similar measurement principle, it can be seen that the sensor studied in the project has high measurement accuracy and low production cost.According to the accuracy evaluation criteria for the soil moisture sensor (RMSE < 0.035 mIn future work, we would like to perform more practical experiments with more different types of soil to verify the measurement accuracy of the proposed sensor. Then, we would be devoted to promoting the application of the pull-out sensor for the portable soil profile moisture measurement of multiple plots with precision agriculture practitioners.


The plasticizer is crucial in the plant-based soft capsule. However, meeting the quality requirements of these capsules with a single plasticizer is challenging. To address this issue, this study first investigated the impact of a plasticizer mixture containing sorbitol and glycerol in varying mass ratios and the performance of the pullulan soft film and capsule. The multiscale analysis demonstrates that the plasticizer mixture exhibits superior effectiveness in enhancing the performance of the pullulan film/capsule compared to a single plasticizer. Furthermore, thermogravimetric analysis, Fourier transform infrared spectroscopy, X-ray diffraction, and scanning electron microscopy indicate that the plasticizer mixture enhances the compatibility and thermal stability of the pullulan films without altering their chemical composition. Among the different mass ratios examined, a 15:15 ratio of sorbitol to glycerol (S/G) is identified as the most optimal, leading to superior physicochemical properties and meeting the requirements for brittleness and disintegration time set by the Chinese Pharmacopoeia. This study provides significant insights into the effect of the plasticizer mixture on the performance of pullulan soft capsules and offers a promising application formula for future use. As one of the traditional dosage forms, a soft capsule is often used to wrap liquid drugs . CompareCurrently, the film-forming agents in the plant-based capsules include three categories: cellulosic fat , Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM) are used to multiscale analyze the characteristics of this pullulan material. The results will reveal the effect of the plasticizer mixture on the performance of the pullulan soft capsule and determine the most reasonable product formula.The material chosen for this study was commercially available food-grade pullulan ; Food-grade κ-carrageenan and ι-carrageenan ; Pure glycerol and sorbitol ; Pure potassium citrate, sodium chloride, and anhydrous calcium chloride .w/w based on weight of water) of sorbitol (S) and glycerol (G) at different mass ratios  was dissolved in distilled water and continuously stirred for 10 min under a water bath heating at 85 °C to prepare a film-forming suspension. Then, pullulan , κ-carrageenan , ι-carrageenan , and potassium citrate  were successively added in the solution and constantly stirred at the same temperature for 1.5 h until the powders completely disappeared. Finally, the solution was cooled to 75 °C and kept for 4 h to remove trapped air bubbles.The plasticizer mixture  was poured onto the plate, which was then dried for 24 h at a relative humidity (RH) of 45% and an ambient temperature of 30 °C. The dried-up films were kept in polyethylene bags for subsequent measurements.The soft capsule was prepared by transferring the above-prepared solution to a soft capsule machine , as seen in The mechanical properties of the films, including elongation at break  and tensile strength , were evaluated using an electronic tensile testing machine , following the procedures specified in the Chinese Pharmacopoeia (11th edition). Specifically, each film sample was cut into a dumbbell shape with a width of 10 mm and a length of 80 mm, ensuring consistency among the samples. The sample was fixed between the device grips with an initial grip separation of 50 mm and cross-head speed of 60 mm/min, with a tensile load cell of 0.5 kN. Each test consisted of three replicate measurements.F, N) by the cross-sectional area . Percentage EB was calculated by dividing the increase in length at break (L) by the initial length (L0) of the sample and multiplying by 100. It can also described in the following equations:TS was calculated by dividing the peak stress (2) and t (h), respectively, represent the test area and the test time.The water vapor permeability (WVP) of the film was determined by adopting a standard cup method . Each fi0. Then, the sample was placed on a polypropylene tray in the prepared glass and was intermittently weighed as mt at 2, 4, 8, 24, 48, 72, and 96 h. The moisture sorption rate (MSR) can be calculated according to Equation (4).The moisture sorption test of the film was carried out according to the previous method . To keepEach test was done in triplicate.−1 under argon.The thermal stability and decomposition characteristics of films were measured using a thermogravimetric analyzer . Each film was cut into small pieces that weighed around 5–10 mg and was put in an alumina crucible. They were heated from 30 °C to 600 °C at a rate of 10 °C/min−1 and a scanning range of 400–4000 cm−1 and 32 scans.Fourier transforms infrared spectroscopy  was used to obtain the FTIR spectra of films. The ground film (1.0 mg) and KBr powders (100 mg) were placed in an agate mortar to prepare pellets under hydraulic pressure. FTIR spectra of each pellet were performed with a resolution of 4 cmThe crystallinity of the film was measured using an X-ray diffractometer . The scanning range of the diffraction angle 2θ° was 5–40°, with a scanning rate of 5°/min.Both the surface and the cross-section of the film were observed by scanning electron microscope . The film was immersed in liquid nitrogen and then fractured into pieces. They were fixed to a sample holder by conductive carbon tape and then sputter-coated with a gold target in a vacuum evaporator. The images were taken at an accelerating voltage of 8 KV.The disintegration test for the soft capsules was conducted following the Chinese Pharmacopoeia. Every time, six soft capsules from a batch of samples were placed in the simulated gastric juice in the disintegration tester  at 37 ± 2 °C. The time all six capsules completely disintegrated was recorded as disintegration time.The brittleness test for the soft capsules was carried out according to previous research . The sofThe reliable mechanical properties of polysaccharide film are crucial to produce high quality soft capsules . TherefoThe performances of the soft capsule in moisture absorption and water vapor transmittance are essential to the storage and application of the capsule. Therefore, the effects of the mass ratio of S/G on moisture absorption and water vapor transmittance of the pullulan soft film are studied, and the results are, respectively, shown in The thermogravimetric analysis (TGA) was used to evaluate the thermal stability of the pullulan soft films at different mass ratios of S/G, and the result is exhibited in −1 represents the stretching vibration of the glycosidic bond [−1 corresponds to the axial deformation of C-H, which may be caused by the absorption of symmetrical and asymmetrical stretching of -CH3 or even groups of -CH2 and -CHO [−1 is prominent due to the formation of hydrogen bonds by the interaction of O-H groups at the end of the pullulan soft film chains and in the plasticizers [−1 and 2920 cm−1, respectively. The peak at 1648 cm−1 in the sorbitol spectrum is attributed to the stretching of the C=O bond. The FTIR spectra of the pullulan films indicate that the peak strengths at 3300 cm−1 and 996 cm−1 were enhanced after the addition of plasticizers, indicating that the addition of plasticizers strengthens the hydroxyl bond and glycosidic bond [The functional groups of the pullulan soft films, without and with plasticizers, pure sorbitol, and pure glycerol, were investigated using Fourier transform infrared spectroscopy (FTIR), and the results of the FTIR spectra of the six materials are shown in dic bond , while tand -CHO . A broadticizers . The maidic bond . MoreoveX-ray diffraction (XRD) is used to investigate the crystal structure of the pullulan soft film. The trends in the XRD patterns of pullulan soft films with pure sorbitol, pure glycerol, and their compounds are shown in The Scanning electron microscopy (SEM) analysis was carried out to characterize the morphology of the surface and cross-section of the pullulan soft films with pure sorbitol, pure glycerol, and their compounds, and the results are shown in According to the requirements of the Chinese Pharmacopoeia (11th edition), one of the critical indicators of the soft capsule is that the soft capsule should be disintegrated within 60 min. Therefore, the pullulan soft capsules with various mass ratios of S/G and storage time were evaluated based on the disintegration time, and the results are shown in The friability of the soft capsule is also a critical indicator of a capsule, so the performance of the pullulan soft capsules with different mass ratios of S/G and storage time are shown in The performances of the pullulan soft film and capsule with the plasticizer mixture of sorbitol and glycerol were studied in this work. The results show that the mixture of plasticizers makes up for the deficiencies of the single plasticizer, improving the quality of the pullulan products. According to the results of TGA, FTIR, XRD, and SEM, it was found that using the plasticizer mixture can improve the thermal stability and compatibility of the pullulan films. The single plasticizer with pure glycerol has a weak connection with the pullulan, improving the fluidity of the pullulan glue, but it reduces the tensile strength of the film. Meanwhile, glycerol has strong hydrophilicity, resulting in poor storage performance of the capsule. The single plasticizer with pure sorbitol improves the crystallinity of the film, resulting in higher tensile strength, but lower elongation at break. Given the excellent elongation at break, temperature stability, and minimal friability exhibited by pullulan products containing a plasticizer mixture of S/G ratio 15:15, along with no significant defects in other characteristics and performance, the soft pullulan capsule with this S/G ratio appears to have strong potential for future applications.


Aim and Objectives: The objective of our retrospective study was to investigate the changes in pNFH levels in cerebrospinal fluid, which is a reliable marker of neuronal damage, after the loading dose of nusinersen in different types of spinal muscular atrophy. Materials and Methods: We analyzed the spinal muscular atrophy types, the number of copies of the SMN2 gene, and the progression of the motor status using specific motor function scales in a group of 38 patients with spinal muscular atrophy types 1, 2, and 3. Results: We found a significant inverse correlation between pNFH levels and patient age, progress on functional motor scales, and nusinersen administration. Our results also revealed that the neurofilament levels in the cerebrospinal fluid were higher in patients with 2 SMN2 copies than those with more than 2 copies, although the association was not statistically significant due to the abnormal distribution of the values. Conclusions: We identified several predictors of favorable evolution under nusinersen treatment, including spinal muscular atrophy type 1, children aged ≤ 30 months, and the presence of only 2 copies of SMN2. Our study provides important insights into the use of pNFH as a biomarker to monitor disease progression and responses to treatment in patients with spinal muscular atrophy. Neurofilaments (NF) are specific neuronal protein heteropolymers, analyzed over time as markers of neuronal degradation in neurodegenerative diseases , which aSpinal muscular atrophy (SMA) is a rare neurodegenerative disease that affects approximately 1 in 10,000 newborns, caused by insufficient synthesis of the survival motor neuron (SMN) protein, which occurs in 95% of cases due to a biallelic deletion in the SMN1 gene 5q13) ,9. About ,9. AbouThe assessment of motor skills is performed using motor functional scales that are adapted to age and are specific to patients with spinal muscular atrophy . The HamIn 2016, the U.S. Food and Drug Administration (FDA) approved Spinraza (nusinersen), the first drug for SMA, a modified antisense oligonucleotide that acts on the expression of the SMN2 gene, leading to the production of a full-length SMN2 transcript . This trWe think that the initiation period can be considered as having the greatest impact on the clinical condition of the patients, with reductions until the disappearance of the destruction of the motor neurons by ensuring the sufficient amount of SMN necessary for their survival. The evolution of the pNFH level during this period is mostly on reducing the damage to the motor neurons due to the treatment.The aim of this study was to analyze the changes in pNFH levels in cerebrospinal fluid after the administration of the loading treatment with nusinersen compared to the initial level of this biomarker, at different ages, with different phenotypes, and in different types of SMA. Additionally, we investigated the relationships between the results obtained on motor function evaluation scales and the evolution of the pNFH levels in cerebrospinal fluid as an expression of the rescue of motor neurons. In this monocentric retrospective study, we examined the level of pNFH neurofilaments in cerebrospinal fluid during the first six months of treatment, which corresponds to the administration of loading doses with nusinersen. The analysis was conducted by collecting two samples, before the initiation of treatment (first sample) and six months after the initiation, at the end of the loading period, just before the administration of the first dose of maintenance treatment (second sample). The results were analyzed based on the age, the SMA type, the number of SMN2 copies, and the evolution on specific motor functional scales.We followed the evolution of 38 genetically confirmed patients diagnosed with SMA who were accepted into the National Treatment Program with Nusinersen between October 2018 and July 2021. To be eligible for the study, the patients had to be under 18 years of age, have bi-allelic pathogenic or likely pathogenic mutations of the SMN1 gene, possess a minimum of two copies of the SMN2 gene, and have a general condition unaffected by any additional acute disease. The exclusion criteria for the study were as follows: the existence of another disease-modifying treatment and the presence of other known diseases that increase the level of neurofilaments, temporary diseases that can influence motor performance, recent acute or chronic inflammatory diseases, other diseases that evolve with acute or chronic neuromotor degeneration, or cerebral or spinal disorders that would interfere with performing a lumbar puncture or influence the circulation of cerebrospinal fluid .The study was approved by the Ethics Committee of the National Teaching Center for Children’s Neurorehabilitation “Dr. Nicolae Robanescu” (agreement numbers 7464/01.10.2018 and 8818/16.12.2020), and all patients had the consent of the authorized legal owners to participate, in accordance with local regulations and the World Medical Association Declaration of Helsinki, revised in 2000, Edinburgh. The legal caregivers of the participants were informed about the purposes of the use of their data, and all patient data were correctly anonymized.The clinical and laboratory assessments, as well as the treatment, were carried out by specialized, trained, and certified personnel, including pediatricians and pediatric neurologists for clinical parameters, physical therapists for functional parameters, and laboratory doctors for biological parameters. All evaluations were made by the same person to avoid the risk of bias in the analysis and the interpretation of the data. After the neurological and pediatric evaluations, qualified staff specialized in assessing children with SMA at the Dr. Nicolae Robanescu National University Center for Children’s Neurorehabilitation evaluated the patients on mobility and physical ability assessment scales recommended according to age and locomotor status for children with SMA , just beBefore the administration of nusinersen, cerebrospinal fluid samples were collected in aseptic conditions through lumbar puncture in an amount of approximately 5 mL according to the administration protocol of the drug . The colThe demographic, anthropometric, motor development milestone achievement, clinical, and laboratory data of the participants were collected from the patient observation sheets and the hospital computer program.t-test for comparisons of the central tendency of the baseline characteristics and a Wilcoxon signed ranks test for continuous variables with an abnormal distribution. The normality of distributions was assessed using the D’Agostino–Pearson test [t-tests [p-value less than 0.05 was considered statistically significant, and all statistical tests were two-tailed [The statistical analyses were conducted using IBM SPSS Statistics 24 and Microsoft Excel 2021 software. The continuous variables were reported as means ± SDs (standard deviations) if variables were uniform distributed or as medians  if the variables were abnormally distributed. In cases of continuous variables with a uniform distribution, we used a paired son test . The Man[t-tests ,23,24 or[t-tests ,26 were [t-tests  was usedo-tailed ,29. We uOf the patients included in the study, 18.42% were diagnosed with SMA type 1 and all had 2 SMN2 copies. SMA type 2 was present in 52.63% of the patients, of which 20% had 2 SMN2 copies, 75% had 3 SMN2 copies, and 5% had 4 SMN2 copies. SMA type 3 was present in 28.95% of the patients, of which 27.27% had 2 SMN2 copies and the remaining 72.73% had 3 SMN2 copies, as shown in There were significant differences in the age at the onset of treatment among the SMA types in the study group, with mean ages ranging from 2.07 to 215 months, with type 1 patients starting treatment at a much younger age than those with types 2 and 3  was determined using the ELISA technique, according to the instructions of the manufacturer of the reagent kit, EUROIMMUN Medizinische Labordiagnostika AG , with the help of the ELISA BIORAD 3100 PSC microplate reader  .The range of pNFH neurofilament levels in the cerebrospinal fluid before treatment was quite wide, ranging from 0.035 ng/dL in a 104-month-old patient with 3 SMN2 copies, diagnosed with SMA type 2, to 3.321 ng/dL in a 3.63-months-old patient diagnosed with SMA type 1 with 2 SMN2 copies, the patient with one of the most severe forms of the disease.No statistically significant correlations were observed between the pNFH level and macroscopic appearance, chlorine ion concentration, or cellularity, neither before the initiation of treatment nor after 6 months of treatment.p < 0.0001, r = −0.588) and SMA type . The pNFH neurofilament level after 6 months of treatment (pNFHf) was moderately correlated with the pNFHi  and weakly correlated with the SMA type  and motor scores before the initiation of treatment . The motor scores after 6 months of treatment were strongly correlated with the motor scores before the initiation of treatment , moderately correlated with the patients’ age , and weakly correlated with the number of SMN2 copies . The SMA types were moderately correlated with the motor scores before the initiation  and the patients’ age  and weakly correlated with the number of SMN2 copies . The patients’ age was moderately correlated with the number of SMN2 copies . The distribution of pNFH neurofilament levels in cerebrospinal fluid, before and 6 months after the initiation of the treatment, according to SMA type, is shown in We identified 12 statistically significant correlations between the analyzed variables. Thus, the pNFH neurofilament level before the initiation of treatment (pNFHi) was moderately correlated with the patients’ age  and two outliers classified as “extreme values” (represented by stars). Additionally, there was one extreme value outlier for SMA type 3. After 6 months of treatment, there were 3 outliers: two extreme values for SMA type 2 and one out value for SMA type 3. The out values are moderately far from the central range of the data, respectively more than 1.5 interquartile range (IQR’s) but less than 3 IQR’s from the upper and lower boundaries of the boxplot, which represent the 75th percentile (Q3) and the 25th percentile (Q1). An outlier is considered to be “out” if it falls above 1.5 times the IQR (the range between Q3 and Q1) but below 3 times the IQR from the end of the box. The extreme value refers to the values that are even farther from the central range of the data, and are considered outliers if they exceed 3 times the IQR. p = 0.08), as shown in the graph provided in The patients with 2 SMN2 copies had a higher level of neurofilaments in the cerebrospinal fluid compared to patients with more than 2 SMN2 copies  and two outliers classified as “extreme values” (represented by stars). After 6 months of treatment, there were two extreme values, one for 2 SMN2 copies and one for 3 SMN2 copies. t-test for SMA types 1 and 3 and the Wilcoxon signed ranks test for SMA type 2:t(6) = 3.848, p = 0.008;SMA type 1: p = 0.012;SMA type 2: z = −2.501, t(10) = 4.088, p = 0.002.SMA type 3: The difference between the pNFH level (ng/mL) before treatment and after 6 months of treatment was statistically significant in terms of the decrease in the amount of neurofilaments in the cerebrospinal fluid after treatment (the 4 loading doses), according to the paired Regression equations for the pNFH levels after treatment (pNFH2) depending on the pNFH levels before treatment (pNFH1) for each type of SMA are shown in t(6) = 4.398, p = 0.005;CHOP INTEND (SMA type 1): t(17) = 2.832, p = 0.012;HFMSE (SMA type 2): t(10) = 3.184, p = 0.01.HFMSE (SMA type 3): Analyzing the motor function evolution after the nusinersen treatment, based on the motor scale assessment scores and using a paired samples test, we found statistically significant improvement in all types of SMA, as shown in p-value of 0.001.For the studied group, from the point of view of evolution on the functional motor scales, the increase in score percentages on the CHOP INTEND and HFMSE scales 6 months from the initiation of nusinersen treatment compared to the initial values relative to the maximum value of the motor scales, 28 patients (73.7%) out of the 38 patients showed a positive evolution from the point of view of increasing the relative score obtained on the motor function evaluation scales 6 months from the initiation of the treatment, 10.5% (4 patients) showed no changes in scores, and 6 patients (15.8%) showed a decrease in motor evolution. These relative percentage increases were between a minimum of –100 and a maximum of 262.5, with an average of 30.83% (SD = 61.5%), a median of 13.92% (IQR = 42.7%), and a In terms of the demographic and clinical characteristics, the study included a diverse group of patients with varying ages and types of SMA, as well as different numbers of SMN2 copies. This allowed for a comprehensive analysis of the relationship between these factors and the level of pNFH in the cerebrospinal fluid, as well as the effectiveness of the nusinersen treatment for motor function. It is important to emphasize that SMA is a rare disease, and the number of patients included in our study is significant for this rare genetic condition. The wide range of pNFH values observed in the study is also notable, with the highest levels of pNFH being determined before the initiation of treatment, in the most serious forms of the disease, and in patients with the lowest scores on the functional motor scales, similar to the results of other studies in the literature ,31,32. Like other studies, the findings show that nusinersen treatment can lead to improvements in motor function and a decrease in pNFH levels in pediatric patients with SMA . The treBased on the non-parametric distribution of the relative percentage increases in the MFM scale scores, we identified a cut-off value of 13.92% for better motor function improvement after 6 months of nusinersen treatment. Upon the analysis of patients with values above the cut-off, we identified the factors that predicted a favorable outcome under nusinersen treatment: diagnosis of type 1 SMA, age of children ≤ 30 months, 2 copies of SMN2 gene, a baseline pNFH neurofilament level > 0.18 ng/mL, a difference between the final and initial pNFH values greater than 0.15 ng/mL, and a percentage decrease in neurofilaments (pNFH) in the cerebrospinal fluid after 6 months of nusinersen treatment of more than 85.99%.We consider that the results of the study are limited by the possibility of the existence of some additional factors, independent of the underlying disease, which could interfere both with the level of pNFH and with the performances on the motor scales, with special roles played by the degree of neuromuscular degradation at the time of treatment initiation and the individual ability to follow physical therapy. Changes in the level of pNFH over time due to treatment with nusinersen can be influenced by the occurrence of episodes of acute diseases between the administered doses, episodes that can evolve with neuronal destruction on the one hand but can also influence the evolution on the functional motor scales by limiting the temporary motor capacity and compliance with the physical therapy program.With the deepening of knowledge about SMA, the evolution of the technology in the field of genetics, the possibility of an early diagnosis, and especially the approval of some disease-modifying treatments, the classification of the types of this rare disease based on functional criteria has become controversial. Being a recently approved treatment, for these patients, the time of initiation of the treatment is not related to the onset of symptoms or the time of diagnosis. After six months of receiving nusinersen treatment, the motor function of most of the patients included in the study showed an improvement, as revealed by an increase in their scores on the motor function scales, while a few of the patients did not show any changes in their scores or experienced a decrease. These findings suggest that the majority of patients experienced a positive response to the nusinersen treatment in terms of increased motor function scores and decreased levels and percentage variation of pNFH in the cerebrospinal fluid. The treatment with nusinersen has been shown to have a high clinical success rate in patients with spinal muscular atrophy, especially for serious forms of the disease, with the treatment initiated as early as possible and with a value as high as possible of the level of pNFH in the CSF at the initiation of the treatment. However, it is important to continue monitoring these patients over time and also study any other groups of patients to check in detail the importance of each of these predictive factors to determine the effectiveness of the treatment in terms of the clinical and motor recovery for a better understanding of the disease and its treatment, and to potentially improve the outcomes for patients with spinal muscular atrophy.


The effects of attapulgite and montmorillonite calcinated at 750 °C for 2 h as supplementary cementing materials (SCMs) on the working properties, mechanical strength, phase composition, morphology, hydration and heat release of ordinary Portland cement (OPC) were studied. The results show that pozzolanic activity increased with time after calcination, and with the increase in content of calcined attapulgite and calcined montmorillonite, the fluidity of cement paste exhibited a downward trend. Meanwhile, the calcined attapulgite had a greater effect on the decrease in the fluidity of cement paste than calcined montmorillonite, and the maximum reduction was 63.3%. Within 28 days, the compressive strength of cement paste with calcined attapulgite and montmorillonite was higher than that of the blank group in the later stage, and the optimum dosages of calcined attapulgite and montmorillonite were 6% and 8%, respectively. In addition, the compressive strength of these samples reached 85 MPa 28 days later. The introduction of calcined attapulgite and montmorillonite increased the polymerization degree of silico-oxygen tetrahedra in C-S-H gels during cement hydration, thereby contributing to accelerating the early hydration process. In addition, the hydration peak of the samples mixed with calcined attapulgite and montmorillonite was advanced, and the peak value was lower than that of the control group. In termH)2·nH2O ,8,9. ThiMg)2SiO4O(OH)2·nH2H)2·nH2O . also stH)2·nH2O . used moH)2·nH2O . The comH)2·nH2O ,15 and eH)2·nH2O ,16, and However, Safa et al.  found thIn this case, there is evidence that it is feasible to use calcination for the treatment of mineral raw materials to improve the performance of cement slurry. Moreover, attapulgite and montmorillonite feature a wide range of sources, low cost, abundant minerals and environmental protection properties and can also reduce the extensive energy consumption of the cement industry, thereby promoting the sustainable development of mineral clay, which is endowed with huge practical significance and great application potential. To this end, on the basis of a large number of experiments, attapulgite and montmorillonite were hereby placed in a Muffle oven and calcined at 750 °C for 2 h. The influence of the calcination mechanism of clay mineral materials on the hydration process of cement was revealed by preparing two calcined clay-cement-based binary composite cementitious materials. The calcined products were added to ordinary Portland cement with different dosages (mass fraction), and the effects of different dosages of calcined products  on the hydration, micro-properties and hydration heat release of ordinary Portland cement were explored.3 was used. The basic mechanical properties of the cement are shown in Herein, ordinary Portland cement (OPC) produced by Huaxin Cement Co, LTD with a relative density of 3100 kg/mThe two clay powders were spread evenly on a porcelain plate. In order to ensure adequate calcination, the proportion of the powder placed on the porcelain plate should not exceed two-thirds. Additionally, complete dihydroxylation of mineral clays requires high calcination temperatures, but too-high calcination temperatures can cause particle sintering . TherefoCalcined or uncalcined clay powder was mixed into the cement according to the mix ratio in Herein, the residual quantity method summarized by Zhong  was adop2 and Al2O3 in the calcined clay powder.Note: The fluidity of the mixtures was determined in accordance with Chinese National Standard GB/T 2419-2005 by measuring the flow diameter.According to GB/T 17671-1999 “Cement Mortar Strength Test Method (ISO Method)”, the compressive performance of the test block was tested, and the samples prepared were placed in the center of the universal testing machine during the test. The compression test was conducted using a YAW4605 pressure testing machine with a running speed of 2.4 kN/s, and the average value of the three samples was taken as the test result.−1 on an infrared spectrometer  using the potassium bromide tablet pressing method. The hydration heat of the samples was analyzed using an isothermal calorimeter  at 25 °C for a duration of 72 h.After the compression strength test, the samples were collected and soaked in absolute ethanol for 24 h to stop hydration. Appropriate bulk samples were selected for micromorphology analysis using scanning electron microscopy . These samples were plated by covering them with gold to make them conductive through the sputtering process in a duration of 90–150 s before the SEM test. The SEM analysis was carried out using a Japanese Hitachi SU8010 machine, the detection resolution, working distance and working voltage of which were fixed at 10 nm, 20–40 mm and 15 KV, respectively. Representative samples at certain hydration ages were crushed and ground using a pestle and mortar for XRD, FTIR and other tests. The composition of raw materials and the phases in hardened cement paste were checked using a D8-Discover X-ray diffractometer  with a CuKα1 radiation, a voltage of 40 kV, a current of 36 mA and a scanning speed of 3°/min. To analyze the molecular bonds and functional groups of the samples, FTIR testing was performed between the wavelengths of 500 to 4000 cm2, Al2O3, MgO, Fe2O3 and CaO, in order of mass fraction, and the mass ratio of the main oxides did not change much after the high-temperature calcination treatment of CAG. For the MT/CMT system, similar to the AG/CAG system, the chemical composition did not change significantly after the high-temperature calcination treatment, and the major oxide components were SiO2, Al2O3, CaO, MgO and Fe2O3 in the order of mass fraction. First, the activity indices of the samples was measured on days 7, 14, 21 and 28, with the data of days 5, 18 and 25 interspersed to ensure the precision and reliability of the data.The chemical composition of the calcined attapulgite and montmorillonite was analyzed and the results are shown in the following 2 and Al2O3 in the calcined clay mineral powder samples reacted with Ca (OH)2 and the reaction products could be dissolved in hydrochloric acid solution. Additionally, CaO and other components in the powder samples could also be dissolved in hydrochloric acid solution. The active component content of sample reaction could be obtained by following this very principle, and the activity rate of the powder was obtained. In this experiment, active samples with different curing days were prepared to explore the reaction degree of powder with different curing periods. The change trend of the pozzolanic activity of these two calcined clays over time is shown in Active SiOAs indicated by It is noteworthy that the environment of the CAG/CMT in the saturated lime aqueous solution was different from that in the cementitious system. The former was in liquid solution, and its ion migration and other obstacles were relatively small, while in the case of the latter, because of the hydration reaction of cement clinker, the generation of hydration products would inevitably wrap and hinder the active reaction of the CAG/CMT powder. Therefore, the effect evaluation could laterally reflect the favorable pozzolanic activity of CAG/CMT, but failed to quantitatively describe the reaction process and amount of CAG/CMT in the cementitious system.After being calcined at 750 °C, the particle-size microstructures of attapulgite and montmorillonite were significantly changed. As indicated by In concrete clay, mineral materials are mainly manifested in the reduction of water-reducing-agent dispersion, reducing the work performance of fresh concrete. Although concrete and cement slurry are very different, they are inextricably linked to each other. Gaining further knowledge about cement slurry is a good guide for concrete batching design and functional application.The effects of different mixtures of attapulgite and montmorillonite on the working properties of cement paste before and after calcination are shown in In the AG/CAG group, the clean cement paste system without CAG could be automatically amortized, and the fluidity reached 245 mm during the test. As the AG content increased to 1 wt%, the fluidity of the cementing material decreased sharply to 60 mm and remained stable then, which was attributed to the fact that AG would absorb a large amount of water-reducing agent in the mixing process so that it could not be well dispersed in the system or play the role of water reducing, and due to the unique pore structure of AG, it would further absorb the free water content of the slurry, thereby resulting in a significant reduction in the fluidity of mixed cement paste .In contrast, in the MT/CMT group, although the addition of MT and CMT would have an impact on the fluidity of the cement paste, the addition of MT had a greater impact on the fluidity of cement paste. When 2 wt% was added, the fluidity of cement pastes almost completely disappeared and decreased to 60 mm, which might be attributed to the intercalation reaction between polycarboxylic acid water-reducing agent and aluminum silicate laminates. The microstructure of MT was lamellar, and polycarboxylic acid water-reducing agent had a high affinity with aluminum silicate layers in the MT structure, which seriously affected the further dispersion of water-reducing agent in the cementitious system, thus reducing its fluidity . The addThe development of compressive strength of the cement paste samples with increase in age and different raw material contents is presented in From the perspective of longitudinal comparison, the control group (FO) had a loose needle-like structure from early curing at Day 3, and the hydration products of these needle-like disordered structures mainly contained calcium carbonate hydrate and a small amount of ettringite hydrate products, while the hydration products at Day 7 could be obviously observed as hexagonal calcium hydroxide (CH) hydrate. In addition, a large number of loose needle-like structures disappeared, the hydration products tended to plate and the structure was denser than that at Day 3. As the curing period was prolonged to 28 days, the tricalcium silicate in the cement clinker was heavily involved in the hydration process, presenting an obvious plate dense structure. It was noteworthy that obvious cracks could be observed at Day 28, and the directional trend of hydration products was speculated to be the cause of the generation of cracks. From the perspective of cross-sectional comparison, the trend of all microstructures was towards the development of densification, with the control group (F0) showing clear needle-like hydration products  from early maintenance at day 3, and the hydration products in the CM10 group showing more of a network-like structure prototype, indicating that the introduction of CMT was able to improve the microfine organization of the early hydration products and properties . At day 3A) reacted chemically with gypsum and other components to produce ettringite (AFt), which provides early strength for cementitious materials; this is the initial hydration reaction of cement. It is worth mentioning that when the entire system is depleted of gypsum, ettringite (AFt) is converted to monosulfide hydrated calcium sulfaluminate (AFm). Subsequently, dicalcium silicate (C2S) as well as tricalcium silicate (C3S) react chemically with water to produce hydration products such as calcium hydroxide (CH) and C-S-H gels, which provide the later strength of the whole system and are the main source of strength of cementitious materials. The diffraction peaks of hydration products such as CH and C-S-H gels became more and more obvious in our XRD patterns, while the diffraction peaks of components such as C3S in the cement clinker gradually weakened.During the exposure of the cement clinker to water, firstly, tricalcium aluminate  at this point gradually disappeared with the progress of the hydration reaction, which was consistent with the theory of the process of cement hydration. In the early hydration process, the main hydration product was ettringite (AFt), and the absorption peak corresponding to -OH in CH was about 3640 cm−1, while those corresponding to H2O were 3420 cm−1 and 1643 cm−1. In addition, ettringite corresponded to the asymmetric stretching vibration of SO42- at the absorption peak around 1110 cm−1, which, due to the corresponding AFt conversion to AFm in the later hydration process, disappeared gradually with the progress of the hydration reaction [−1 and 874 cm−1 corresponded to Si-O in silicate, while the absorption peak at 1424 cm−1 corresponded to carbonate, which might be caused by the inevitable carbonization reaction during sample preparation or hydration.Infrared analysis can provide more information about the hydration products in a cementitious system. The infrared spectra of the samples at different ages are shown in reaction . The abs−1, indicating that the introduction of CAG contributed to the formation of AFt in the early stage and its transformation in the later hydration process. Additionally, the absorption peak around 900–1000 cm−1 tended to migrate to the higher wave range with the increase in the hydration reaction time and the CAG dosage, suggesting that the Si-O bond gradually polymerized into a Si-O-Si bond [−1, indicating that the introduction of CMT was conducive to the formation of AFt in the early stage and conversion in the later hydration process. Additionally, absorption peaks around 900–1000 cm−1 tended to migrate to higher wave segments with the increase in the hydration reaction time and the CMT content, which also indicated that Si-O bonds polymerized gradually to Si-O-Si bonds, and the degree of silico-oxygen tetrahedron polymerization in C-S-H increased. In addition, there were similar phenomena in the IR spectra of CAG-OPC, and the transformation was speculated to be related to pH reduction in the system [It should be noted that as shown in -Si bond . Similare system .2, Al2O3 and other components in the whole cementitious system to participate in the hydration reaction, thus accelerating the reaction, similar to the study by Kaminskas et al. [The hydration heat was measured at room temperature, as shown in s et al. . Howevers et al. . Figure s et al. .Herein, the pozzolanic activity of attapulgite and montmorillonite was enhanced using calcination, and a series of mixtures was prepared by replacing OPC with calcined attapulgite and montmorillonite. The hydration products and basic properties of these mixtures were correspondingly studied, and the following conclusions were drawn based on the results and analysis:(1) After calcination, the pozzolanic activities of CAG and CMT were significantly enhanced with time, and the addition of CAG and CMT seriously reduced the fluidity of the mixed cement paste, achieving a maximum reduction of 75.5%. Additionally, it was found through testing the mechanical strength of the samples that adding a trace amount of calcined raw materials increased their compressive strength in the late hydration stage;(2) Both the introduction of CAG and CMT contributed to the formation of AFt in the early stage and conversion in the later hydration process, which promoted the gradual polymerization of Si-O bonds to Si-O-Si bonds and increased the degree of silico-oxygen tetrahedron polymerization in C-S-H. The difference between the two was that CAG highlighted the peak generated by secondary ettringite, while CAG did not have a peak generated by secondary ettringite;(3) Given that additional hydration products were generated on the surface of CAG/CMT, hydration in the acceleration period was advanced with the increase in content of the calcined raw materials. However, simultaneously, after the addition of the calcined raw materials, since there was more replacement of the original cement clinker, the cumulative heat release was reduced, which is of great significance to limit the early thermal cracking of concrete buildings.