To evaluate predictability, stability, efficacy, and safety of transepithelial photorefractive keratectomy (TPRK) using smart pulse technology (SPT) (SmartSurface procedure) of Schwind Amaris with mitomycin C for correction of post small incision lenticule extraction (SMILE) myopic residual refractive errors.This study is a prospective, non-comparative case series conducted at a private eye centre in Ismailia, Egypt, on eyes with post-SMILE myopic residual refractive errors because of undercorrection or suction loss  with myopia or myopic astigmatism. The patients were followed up post-SMILE for six months before the SmartSurface procedure, and then they were followed up for one year after that. TPRK were performed using Amaris excimer laser at 500 kHz. The main outcomes included refractive predictability, stability, efficacy, safety and any reported complications.P < 0.0001). Uncorrected distance visual acuity (UDVA) (measured by Snellen's chart and averaged in logMAR units) was improved significantly to 0.1 ± 0.07 (P < 0.0001). UDVA was 0.2 logMAR or better in 100% of the eyes, 0.1 logMAR or better in 91.2% of the eyes, and 0.0 logMAR in 20.6% of the eyes. Corrected distance visual acuity (CDVA) remained unchanged in 79.4% of eyes. 14.7% of eyes gained one line of CDVA (Snellen). 5.9% of eyes gained two lines of CDVA (Snellen).This study included 68 eyes of 40 patients out of 1920 total eyes (3.5%) with post-SMILE technique myopic residual refractive errors. The average duration between the SMILE surgery and TPRK was 6.7 ± 0.4 months (range 6 to 8 months). The mean refractive spherical equivalent (SE) was within ± 0.50 D of plano correction in 100% of the eyes at 12 months post-TPRK. Astigmatism of < 0.50 D was achieved in 100% of the eyes. The mean of the residual SE error showed statistically significant improvement from preoperative − 1.42 ± 0.52 D to 0.23 ± 0.10 D (Transepithelial photorefractive keratectomy using smart pulse technology with mitomycin C enhancement after SMILE is a safe, predictable, stable, and effective technique. Small incision lenticule extraction (SMILE) technique was introduced to preserve biomechanical properties of the cornea, reduce injury to corneal nerves, which results in earlier recovery of postoperative corneal sensitivity, and thus lower incidence of dry eye when compared with laser-assisted in situ keratomileusis (LASIK) . In seveThe most common complication of any refractive procedure is residual refraction. Overcorrection, undercorrection, induced or residual astigmatism are all included; however, refractive regression is characterised as the gradual, partial, or complete loss of initial correction, limiting the predictability, efficiency, and long-term stability of this refractive procedure , 4.Retreatment rates after SMILE have not been extensively reported ; ReinsteThere are many alternatives to choose to correct post-SMILE residual refractive errors or post-SMILE myopic regression . PhotoreThis study is a prospective, non-comparative case series of 68 eyes in 40 patients conducted at a private eye centre in Ismailia, Egypt. All patients provided written informed consent for the procedure and inclusion in this study. This study adheres to the tenets of the Declaration of Helsinki. It was reviewed and approved by the Faculty of Medicine, Suez Canal University Research Ethics Committee. The patients were operated on, examined, and followed up by one surgeon (AAG).Patients underwent SmartSurface treatment, which was a combination of TPRK using the SPT. Inclusion criteria were patients with post-SMILE myopic residual refractive errors because of undercorrection or suction loss  including residual and/or consecutive astigmatism ≥ 0.5 D. The patients were followed up post-SMILE for six months before TPRK. Other inclusion criteria were patients with intraocular pressure (IOP) < 21 mmHg, sufficient corneal thickness to leave > 60% of the original total corneal thickness, and regular corneal topography pattern . Patients were excluded if they had consecutive hyperopia with or without residual or consecutive astigmatism, diabetes mellitus, autoimmune diseases, corneal scars, or ectasia. Patients with insufficient follow-up were excluded from the study.Preoperatively, patients experienced standard eye examinations including slit-lamp examination, indirect fundoscopy, and refraction (cycloplegic and manifest), IOP, and biomechanical properties of the cornea  and corneal resistance factor (CRF)) by ocular response analyser II . Uncorrected distance visual acuity (UDVA) and corrected distance visual acuity (CDVA) measured by Snellen's chart (logMAR units) was recorded. SMILE technique was performed using refractive lenticule extraction (ReLEx®) on the VisuMax 500 kHz femtosecond laser system . Attempted cap thickness was set at 110 ± 10 μm  and exceeded the lenticule diameter by 1.0 to 2.0 mm. The size of the lenticule ranged from 6.0 to 7.0 mm, with no transition zone for spherical errors and a 0.1 mm transition zone for astigmatism. It was adjusted according to the mesopic pupil diameter of the patients. The entering incision varied from 3.0 to 4.0 mm. Postoperative follow-up visits were on the 1st day, 1st week, 1st month, 3rd month, and 6th month. Follow-up visits included full ophthalmic examinations, cycloplegic and manifest refraction, corneal topography, UDVA and CDVA, biomechanical properties of the cornea by ocular response analyser II, and slit-lamp examination.All the treatments were done using Amaris excimer laser at 500 kHz with plano target, aspheric, and non-wavefront-guided profiles. TPRK were performed only when the estimated residual stromal thickness was greater than 300 μm. After povidone-iodine scrub of the eyelids and application of skin topical anaesthesia with 0.4% benoxinate hydrochloride drops at 5-min intervals, a wet sponge by balanced salt solution (BSS) was used before laser ablation to uniformly wipe the corneal surface, preventing uneven wetting and, accordingly, uneven ablation. The planning software calculated the size of the optimal transition zone for each treatment, depending on the preoperative refraction and optical treatment zone. The enhancement was ablated by TPRK mode using smart SPT (SmartSurface TPRK). The epithelial ablation profile software uses a central epithelium ablation of 55 µm at the centre and 65 µm at 4 mm. MMC (0.02%)  was applied to the stromal bed for 30 s by a wet sponge. The surface was irrigated with BSS and dried. Then, a soft contact lens  was added, and one drop of 0.3% Tobramycin and 0.1% Dexamethasone was instilled . The patients were advised to use 0.5% moxifloxacin hydrochloride  qid till re-epithelization, remove contact lens, and apply a combination of 0.3% topical Tobramycin and 0.1% Dexamethasone qid for the first week, then decrease to once a week, and 0.3% sodium hyaluronate drops  qid for 4 months.Postoperatively, the patients were examined on the 1st day, 1st week, 1st, 3rd, 6th, and 12th month. Patients’ data were reported at 1, 3, 6, and 12 months postoperatively. Patients were assessed by complete ocular examinations, IOP, CH, CRF by ocular response analyser II, corneal topography, refraction (cycloplegic and manifest), CDVA and UDVA. Complications were recorded. According to Fante's classification , the corThe main outcomes included refractive predictability, which was assessed by the percentage of eyes < 0.5 D of the target correction using the SE. Stability was assessed by comparing postoperative cycloplegic refractions SE at follow-up times. Efficacy was assessed by the percentage of postoperative UDVA to the preoperative CDVA and the efficacy index was the ratio of the mean postoperative UDVA to the mean preoperative CDVA. Postoperative safety was assessed by the percentage of the eyes that gained/lost lines compared with preoperative CDVA; the safety index was the ratio of the mean postoperative CDVA to the mean preoperative CDVA and reported complications.. P-value  of less than 0.05 was considered statistically significant. Graphs were created using either SPSS or Microsoft Excel 2016.Data were coded, entered, and analysed using SPSS . Baseline data of the study population were presented as mean values, standard deviations, and range. Predicted (attempted) postoperative SE refraction was calculated using simple regression analysis, while the mean error in the treatment was calculated as the difference between the attempted and achieved postoperative SE refraction up to the 12th month after surgery. Analysis of visual acuity results was performed by calculating the geometric mean with a standard deviation into logMAR format from Snellen examination resultsThis study included 68 eyes of 40 patients out of 1920 total eyes (3.5%) with post-SMILE myopic residual refractive errors for refractive enhancement. TPRK treatment was bilateral in 28 (70%) patients and unilateral in 12 (30%) patients (8 patients in the right eye and 4 patients in the left eye). Twenty-eight eyes were of male patients and 40 eyes of female patients. The average duration between SMILE surgery and TPRK was 6.7 ± 0.4 months (range 6 to 8 months). The demographic data of the SMILE and TPRK groups are presented in Table At the 12th month post-TPRK, the mean refractive spherical equivalent (MRSE) was within ± 0.50 D of plano correction in 100% of eyes (68 eyes). The distribution of MRSE before and after TPRK can be found in Fig. The mean refractive astigmatism was within 0.50 D and this was achieved in 100% of eyes Fig. . Pre-TPRP < 0.0001) from pre-TPRK − 1.42 ± 0.52 D (range − 0.75 to − 2.50 D) to + 0.35 ± 0.18 D (range + 0.50 to − 0.50 D) at the 1st month, + 0.25 ± 0.27 D (range + 0.50 to − 0.25 D) at the 3rd month, + 0.23 ± 0.12 D (range + 0.25 to − 0.25 D) at the 6th month, and + 0.22 ± 0.11 D (range + 0.25 to 0.00 D) at the 12th month. There was a statistically significant improvement of the refraction up to the third post-TPRK month, then the stability of the refractions up to the 12th month  significantly improved at the 1st, 3rd, 6th, and the 12th month to 0.15 ± 0.08 (range 0.25 to − 0.10), 0.10 ± 0.10 (range 0.2 to − 0.1), 0.10 ± 0.07 (range 0.18 to − 0.10), and 0.10 ± 0.06 (range 0.17 to − 0.10), respectively . Ten eyes (14.7%) gained one line of CDVA (Snellen). Four eyes (5.9%) gained two lines of CDVA (Snellen). No eye had lost line of CDVA . Post-TPRK mean CH at the 1st, 3rd, 6th, and 12th month was 8.49 ± 1.50 (range 7.36 to 10.84), 8.48 ± 1.70 (range 7.36 to 10.84), 8.47 ± 1.60 (range 7.30 to 10.70), and 8.46 ± 1.57 (range 7.29 to 10.60), respectively. There was no significant change in CH during the follow-up period . Post-TPRK mean CRF at the 1st, 3rd, 6th, and 12th month was 9.05 ± 1.80 (range 7.38 to 10.90), 9.01 ± 1.40 (range 7.38 to 10.90), 8.97 ± 1.50 (range 7.28 to 10.90), and 8.96 ± 1.50 (range 7.29 to 10.87), respectively. No significant change in CRF was observed over the follow-up period  with no intraoperative complications. Grade 1 corneal haze was observed in 18 eyes (26.4%) and grade 2 in 4 eyes (6%) but disappeared during the first month postoperatively. Corneal haze grade 2 observed in eyes disappeared by the 3rd month postoperatively. Corneal ectasia was not detected in any of the eyes of the study. No other complications were recorded with complete healing of the epithelium within the first week postoperatively.In this study, predictability, stability, efficacy, and safety of TPRK using SPT (SmartSurface procedure) of Schwind Amaris with MMC for correction of post SMILE myopic residual refractive errors were evaluated. All the treatments were done using Amaris excimer laser at 500 kHz with plano target, aspheric, and non-wavefront-guided profiles. TPRK mode has a higher laser cutting frequency than traditional PRK , many reThe retreatment rate of 3.5% reported in this study is in tandem with previous studies. Siedlecki et al.  reportedIn terms of efficacy, stability, predictability, and safety; refraction significantly improved at the 1st week and remained stable until the end of the 12th month of the follow-up time. Mean UDVA significantly improved from 0.62 to 0.1 logMAR and 100% of eyes were within ± 0.50 D of target refraction. At the 12th month post-TPRK, the efficacy index was 1.04 ± 0.17. A proportion of 14.7% of eyes gained at least one line of CDVA, 5.9% of eyes gained two lines, and CDVA remained unchanged in 79.4% of eyes (n = 54). The safety index was 1.02 ± 0.03 at the 12th month. These data are comparable to the results of surface ablation on virgin eyes. At the end of the follow-up period, the cycloplegic refraction was slightly toward the hyperopic side (mean + 0.22 ± 0.11 D). In this situation, the effects of MMC on overcorrection reported by Leccisotti  have to In contrast to LASIK, which offers flap re-lifting as a safe choice for enhancement, selecting the ideal enhancement method after SMILE remains a highly debatable issue . It may Some limitations should be considered while reading this study. In this study cohort, we have not analysed corneal aberrations and used a non-wavefront aspherical profile for all subjects. Further studies will show if some cases will benefit from corneal wavefront treatment after SMILE as reported by de Ortueta et al. .Transepithelial photorefractive keratectomy using smart pulse technology (SmartSurface procedure) with MMC enhancement after SMILE is a safe and effective technique with high predictability and stability that could correct post-SMILE myopic residual refractive errors and improve both CDVA and UCVA. This avoids flap-related complications caused by the manipulation of the primary SMILE cap.


There are many Western reports on factors influencing coital frequency among men. However, no articles could be found about the factors influencing sexual activity among Chinese men.The aim of this study was to identify the factors that influence the coital frequency of Chinese men.The main outcome measures included self-reported monthly coital frequency, age, occupation, education level, andrology-related scales and dietary habits.Data for 1,407 men aged 18–79 years were collected in the Health Management Center of the Third Xiangya Hospital of Central South University from January 2019 to May 2019. The respondents completed the questionnaires independently or with the help of an interviewer (who read or explained the questionnaires to them) to analyse the factors that influence coital frequency.P = 0.004), IIEF-5 scores (P <0.001), EHSs (P <0.001) and frequency of milk consumption (P = 0.001) were associated with more frequent sexual activity. These statistical associations did not change after further adjustment for age, occupation, and reproductive history. We observed that the frequency of sexual activity showed an increasing trend with a greater number of children, higher IIEF-5 scores, higher EHSs and greater frequency of milk consumption . Both univariate and multivariate analysis results indicated that the frequency of sexual activity decreased with increasing age .In the previous 6 months, the sample had a mean monthly coital frequency (±SD) of 4.34 ± 3.18. Univariate logistic regression results indicated that the number of children (Xiang Y, Peng J, Yang J, et al. What Influences Coital Frequency Among Chinese Men?: A Cross-Sectional Study. Sex Med 2021;9:100363.The coital frequency of Chinese men is associated with erectile function, anthropometric parameters, age, occupation, and dietary habits.  The factors considered include anthropometric parameters,Men participated in a physical examination at the Health Management Center of the Third Xiangya Hospital of Central South University from January 2019 to May 2019. They gave informed consent and could interrupt or withdraw from the interviews at any moment. The inclusion criteria were as follows: (i) 18-80 years old; (ii) underwent complete health check-ups in the Third Xiangya Hospital of Central South University; (iii) physically active and healthy at the time of the study; and (iv) willing to participate in this study.According to the above standards, 1407 consecutive heterosexual men participated in this study. The age of the subjects was 18−79 years old, with an average age of 38.7±9.4 years. All protocols were approved by the Institutional review board of the Third Xiangya Hospital of Central South University (No. 2019-S252).The questionnaire included the average monthly coital frequency in the past 6 months, personal history , IIEF-5 questionnaire, EHS questionnaire, and the premature ejaculation diagnostic tool (PEDT). Finally, the relevant physical examination data of the subjects were imported.The data on the frequency of sexual activity are based on the average frequency of sexual activity per month in the past 6 months, as reported by the subjects. Moreover, the subjects also reported their range of sexual intercourse frequency: none, 1–4 times/month, 5–8 times/month, and ≥9 times/month.Sexual function was assessed by Andrology-related scales, including the IIEF-5, EHS, and PEDT. Erectile dysfunction was assessed by IIEF-5 scores: severe (1–7), moderate (8–11), mild to moderate (12–16), mild (17–21), and no erectile dysfunction (ED) (22–25). The erection hardness was evaluated using the EHS: penis is larger but not hard (i), penis is hard but not hard enough for penetration (ii), penis is hard enough for penetration but not completely hard (iii), and penis is completely hard and fully rigid (iv). Premature ejaculation (PE) was diagnosed by the PEDT: PE (≥11), suspected PE (9–10), and non-PE (≤8).P<0.05 was considered significant.All of the questionnaire results were input into the computer after correction; Microsoft EXCEL 2016 version was used to establish a database. Statistical analyses were performed by utilizing SAS statistical software 9.2 . Means ± standard deviations were used to describe the quantitative indicators, and frequencies were used to describe the count data. The Cochran-Mantel-Haenszel (CMH) test was used to analyze the coital frequency distribution at different levels , and 4. P <0.001), occupation (P = 0.01), reproductive history (P <0.001), waist-to-hip ratio (WHR) (P = 0.008), and BMI (P = 0.0143). The highest levels of coital activity were reported by men who were 30–39 years of age (4.9±3.3), were self-employed (4.97±3.23), had two or more children (4.7±3.07), had a higher WHR (4.47±3.13), and had a BMI reflecting obesity (4.85±3.41).The main anthropometric characteristics of the study population are presented in P < 0.001)  .Table 2TP <0.001) and EHS (P = 0.0057) questionnaires. Those with normal IIEF-5 scores had a mean sexual activity frequency of 5.14±3.47 times/month, and those with EHSs of 4 had a frequency of 4.55±3.36 times/month. In other words, men with normal erectile function have more frequent intercourse.P = 0.0056), the frequency of milk consumption (P = 0.0233), the frequency of fish or seafood consumption (P = 0.0007), and the frequency of fruit consumption (P = 0.0158). Sexual activity was more frequent among men who consumed more fibre and rice (4.48±3.29), who drank milk weekly/regularly/daily (4.69±3.49), who ate fish or seafood weekly/regularly/daily (4.53±2.8) and who ate fruit weekly/regularly/daily (4.64±3.27).In addition, we also studied the relationship between lifestyle factors and coital frequency . Coital P = 0.004), the IIEF-5 scores (P <0.001), the EHSs (P <0.001) and the frequency of milk consumption (P = 0.001) were related to an increased frequency of sex. This was confirmed by multivariate analysis after further adjustment for age, occupation, and reproductive history. The number of children was associated with an increased risk of higher sexual frequency, with ORNO=1=2.944  and ORNO≥2=3.307  compared with subjects without children. We observed the same effect of other factors, such as the IIEF-5 score, EHS and frequency of milk consumption.The mean (±SD) coital frequency per month was 4.34±3.18. P<0.05). Both univariate and multivariate analysis results demonstrated that age was associated with the frequency of intercourse (P<0.001). The test for trend analysis indicated that coital frequency had a decreasing trend with increasing age (P<0.001). However, the average monthly sexual frequency in the 18–30 age group was lower than that in the 30–50 age group. Interestingly, although WHR was not significantly associated with sexual frequency in the univariate analysis, we observed a significant association in the multivariate analysis. Men with an increased WHR (4.47±3.13) had a significantly higher coital frequency than those with a normal WHR (4.2±3.3).The test for trend analysis indicated that with a higher number of children, a higher IIEF-5 score, a higher EHS, and a greater frequency of milk consumption, sexual frequency showed an increasing trend . However, the sex frequency of the 18- to 30-year-old male population was lower than that of the 30−50-year-old male population, which can be attributed to the lack of stable marriage partners. It has been proven that sexual activity increases with marriage.The results of this investigation demonstrate that the mean frequency of sexual activity was 3.8±4.1 times/month among men 18−29 years old, 4.9±3.3 times/month among men 30−39 years old, 4.6±2.5 times/month among men 40−49 years old, and 2.8±2.1 times/month among men over 50 years old. Previous studies have proven that sexual activity tends to decline with age.,,The frequency of sex was higher among men with a BMI indicating obesity (4.85±3.41) and an increased WHR (4.47±3.13). The results of the multivariate analysis found that the frequency of sex increases with WHR. However, Western studies have reported that body mass index showed a trend towards a negative association with sexual frequency for men,P<0.05). Men with two or more children had a higher frequency of sex (4.7±3.07) than other men. Donnelly found a positive correlation between the number of children and the frequency of sex, which is consistent with our results.Self-employed (4.97±3.23) men have more active sex lives than men in other occupations. It is generally noted that working couples have less time to engage in intimate contact, and married couples in which both partners hold full-time jobs engage in sex less frequently than those in which one partner does not work full time outside the home.P = 0.0056), the frequency of fish or seafood consumption (P = 0.0007), and the frequency of fruit consumption (P = 0.0158). Sexual activity was more frequent among men who consumed fibre and rice (4.48±3.29), drank milk weekly/regularly/daily (4.69±3.49), ate fish or seafood weekly/regularly/daily (4.53±2.8) and ate fruit weekly/regularly/daily (4.64±3.27). Moreover, the test for trend analysis demonstrated that sexual frequency increased with the frequency of milk consumption (P<0.05). It can be seen from these results that these men follow regular diets and pay attention to dietary health and dietary diversity. Previous studies proved that adherence to the Mediterranean diet  also improved the frequency of sexual intercourse and significantly reduced the prevalence of sexual dysfunction,,The relationship between lifestyle factors and coital frequency  indicateThese data come from a cross-sectional study, and there was no assessment of differences in factors such as changes over time in lifestyle habits, weight, and occupation. As mentioned above, mental status and marital status between partners will affect male sexual activity. Unfortunately, this study did not evaluate psychiatric diagnosis/interventions and marital status. Statistical significance of this study is set at 5% there is a 1 in 20 chance that each comparison will be spurious and driven by chance alone. Given the number of comparisons in this manuscript there is almost certainly at least 1 if not more type 1 errors.At present, there is a lack of studies on factors related to the frequency of intercourse among Chinese men. In the current study, we found that the coital frequency of Chinese men was associated with erectile function, anthropometric parameters, age, occupation and dietary habits. Therefore, large-scale studies worldwide on larger representative samples are necessary, and more data are needed from other cultures and different age groups.Yali Xiang: Conception and Design, Acquisition of Data; Dongjie Li: Conception and Design, Drafting the Article, Revising It for Intellectual Content; Jingxuan Peng: Analysis and Interpretation of Data; Yuxin Tang: Analysis and Interpretation of Data; Jianfu Yang: Analysis and Interpretation of Data; Jingxuan Peng: Drafting the Article; Yuxin Tang: Final Approval of the Completed Article.


A higher concentration of Se resulted in declined survival of embryos. Accumulation of Se in the body of hatch increased with the increase of element concentration in the aqueous environment during egg swelling. The survival of fry was similar in all groups, while the fry length and weight correlated positively with Se concentration in its body. In Se-poor areas, immersion of fertilised eggs in water enriched with Se during egg swelling can be used to supplement the deficient element to fish in order to improve breeding outcomes in aquaculture.Selenium-poor areas are defined in the Earth. Adequate selenium (Se) consumption enhances the health and growth of organisms, but overdose of it can be harmful and pathogenic. The objective of the study was to investigate Se supplementation effects in the non-feeding stages of fish. Fertilised eggs of sea trout were immersed in water enriched with Se during the egg-swelling period. Se at a concentration from 0.5 to 8 mg Se L2) at concentrations from 0 to 32 mg Se L−1 to find the optimal and toxic dose of Se on early fish development. Se accumulated in the body, embryos’ survival rate, and growth in the first four months of life was examined. Swelling of fertilised eggs in water supplemented with Se at a concentration from 0.5 to 8 mg Se L−1 was associated with a slightly positive impact on the hatching rate. At higher Se concentration, a harmful effect on the survival of the embryo was observed. The survival of fry was similar in all groups, while the fry length and weight correlated positively with Se concentration in its body. Immersion of fertilised eggs in water enriched with Se during egg swelling can constitute a method to supplement the element to non-feeding stages of fish. In selenium-poor areas, this innovative method can be implemented in aquaculture to improve breeding outcomes. Se concentration should be adjusted to the chemical compound, fish species, and Se’s content in the yolk.Adequate selenium (Se) availability enhances the health and growth of organisms, but overdose of it can be harmful and pathogenic. The study’s objective was to analyse the impact of short-term exposure of sea trout fertilised eggs to inorganic selenium (SeO Selenium is a vital trace element incorporated into selenoproteins, which have an extensive impact on the organism. A suitable supply of selenium to the organism is vital for normal functioning ,2. The eAn excessive status of selenium in the body is toxic, resulting in impaired enzyme function and disruption of protein structure ,24,25. I−1) [−1 for drinking water) [−1 of organic selenium in feed to ensure consumer safety. In adult fish selenium requirement for optimal growth is higher [In the environment, selenium is found in different oxidation states in organic and inorganic compounds. Inorganic forms as elemental selenium, selenite and selenides exhibit low uptake—organic forms of selenium, such as selenomethionine, selenocysteine, and methylselenocysteine exhibit higher bioavailability ,38,39. G−1)  or Worldg water)  except ig water) ,43,44,45g water) ,46,47,48g water) ,51,52,53g water) ,54. The g water)  recommens higher ,57,58.Salmo trutta m. trutta L.) is a migrating salmonid fish. Adult fish leave marine waters and enter rivers to spawn. The natural range of occurrence of the species includes Atlantic, North, White, and Baltic Sea basins. In Poland, the sea trout is an economically important species due to the extinction of the Atlantic salmon in the 1980s [−1 [Sea trout  entering the Rega River for spawning (NW Poland) were purchased from Polish Fishing Association division Szczecin. The eggs and milt originated from three individuals of each sex. After transport (4 °C) to the laboratory and assessing milt ) and eggs’ quality, the gametes were mixed in equal proportions from different individuals in order to obtain fertilisation. After rinsing the fertilised eggs with aquarium water from a closed-circuit, they were divided into eight groups of 150 eggs, each in three repetitions, each containing about 50 eggs. Se content in unfertilised egg and hatchery water was very low (selenium-poor area). In egg, the element concentration was 1.6 µg Se g−1 (dw)  were prepared to dissolve SeO2 (Merck) in separated aquarium water. Fifty fertilised eggs were placed in 100 mL of selenium solution for 3 h to swell at 6 °C. After the swelling, the eggs were rinsed with separated aquarium water and transferred to baskets floating in the basins (three repetitions) with a closed water circuit. The survival rate was monitored by counting and removing white eggs. Hatching time was similar in all groups between February 18 and 27. Three weeks after hatching, fish from selected variants  was reared in basins of 300 L to the age of four months. Initially, the water temperature was kept at 6 °C; then, it gradually increased in subsequent months to 16 °C. Water flow was 300 L h−1. Chemical and physical parameters of water were recorded three times a day. During rearing, N-NH3 mean concentration was 0.007 mg L−1 ± 0.003, diluted oxygen 7.20 mg L−1 ± 0.41, and pH 7.41 ± 0.82, photoperiod typical of this season. Fry was fed with frozen foods, adult Daphnia 1.13 µg Se g−1 (dw), Chironomidae larvae 0.29 µg Se g−1 (dw), and granulated feed (Skretting. Perla Larva Proaktive 4.0) containing 0.23 µg Se g−1 (dw). Feeding time takes place between 7.00 a.m.–5.00 p.m. every 3–4 h.Eggs and milt of wild sea trout  and F) were taken using callipers with an accuracy of 0.1 mm. After the measurements, fish were released to the basin. Condition factor CF was calculated using the following formula CF = 100 × W × LF−3. According to Polish law, permission from the Ethics Committee for Animal Experimentation (no. 86/2016) was obtained to conduct the experiment.At monthly intervals from the beginning of feeding, 30 individuals were collected from each basin for morphometric testing from March through June. After short-term anaesthesia in MS-222, measurements of fish body weight (W) with accuracy to 0.1 mg and length  on a VARIAN SpectrAA 220 FS apparatus . The methodology of Diaz-Alarcon et al. [The concentration of accumulated selenium was determined in wet weight (ww) in the hatch and four-month-old fry euthanised by MS-222 overdose and stunning with the destruction of the brain. For the purpose of mineralisation, 2 g of wet material was collected. The material was mineralised in a MARS-5 microwave oven by CAM . In order to check the accuracy of the method, selenium content was determined in a reference material—DOLT-2 from the National Research Council of Canada. Selenium reference content was 6060 ± 409 μg kgn et al.  was utilShapiro–Wilk test of normality of the analysed data and Levene variance homogeneity test were conducted. One-way ANOVAs was used to test the significance of differences in selenium content accumulated in hatching and four-month-old fry between the experimental groups. Two-way repeated-measures ANOVA was used to analyse the significance of differences in length, weight, condition factor, and intestinal and renal epithelial cell height between the experimental groups. Fisher’s least significant difference test was used for post hoc comparisons. A chi-squared test was used to compare the frequencies of fry survival. Pearson’s correlation coefficients were calculated to evaluate the relationship between selenium concentration in the fish body and fish length and weight. All analyses were performed at a significance level of 0.05 using the Statistica v. 13.1 software . The data are presented as means ± SD.p > 0.05) . The survival rate in this variant at cleavage, gastrulation, eyed-egg, and hatching stage was 100 ± 0%, 90.4 ± 4.5%, 82.5 ± 10.4%, and 79.0 ± 10.3%, respectively  . The incectively .−1) was lethal for embryos. In this group, higher mortality of 11.0 ± 4.0% at the cleavage stage was detected (p < 0.05). During gastrulation, in this group, mortality was very high (mean 92.4 ± 5.7%); at eyed-egg, all of the eggs died (LT100) (The highest selenium amount added to the water (32 mg Se L (LT100) .None of the macroscopically detected body deformations was found in hatched larvae in the control sample. In experimental groups, singular cases of spine deformities were observed; they comprised 0.3% of individuals in all experimental variants.−1 (dw), whereas, in the 16 Se group, selenium concentration was 5.4 ± 0.3 µg g−1 (p < 0.05) (Selenium concentration accumulated in the hatch body increased with increasing the element concentration in the aqueous environment during egg swelling . Detecti < 0.05) .−1 (dw) (p < 0.05)  .p > 0.05), whereas the growth of fry differed between variants (p < 0.05). In the group of fish with higher selenium content in the body, greater length and weight were recorded (p < 0.05). The mean body length of one-month-old fry in the control group was lowest on average of 25.12 ± 0.57 mm. Statistically, significantly greater lengths were observed in samples 8 and 16 Se hatching, which attained 26.47 ± 0.81 mm and 26.73 ± 1.21 mm, respectively (p < 0.05). In the control sample, hatching weight was on average 0.09 ± 0.01 g. In contrast, in variants 2, 8 and 16 Se it was significantly higher, and it was 0.12 ± 0.01 g, 0.12 ± 0.01 g, and 0.13 ± 0.01 g, respectively (p < 0.05). The condition coefficient was similar between the experimental groups; in the four-month-old fry, it remained in the range between 0.72–0.75.The survival rate of fry to the age of four months did not differ between control and experimental samples; it remained in the range from 79.8 to 86.2%  compared to the control group . The optimal selenium content in the body also had a slightly positive effect on reducing egg mortality and hatching rate (the effect was non-significant). These features also confirm the important role of selenium in fish condition and survival [Selenium is an essential nutrient for organisms but is toxic at high levels ,28,29,30survival ,14,19,212 at a concentration of 16 mg Se L−1 was associated with a significantly lower hatching rate when Se content in the hatch body was 5.4 µg Se g−1 (dw). Our results confirm that small differences between the appropriate and harmful concentrations of Se occurred [100). The experiment by Kowalska-Góralska [−1 in the hatch body resulted in a slightly negative impact on the hatching rate. The threshold is similar to that obtained in the current study. Some authors indicated a somewhat lower threshold of >4 µg Se g−1 than mentioned by us above [−1 (dw), respectively [−1. The large differences in Se concentrations disclosed in papers are probably related to the age of the fish, their ability to bioaccumulate this element, and the species.It is known that selenium overdose is harmful, and we observed a minor difference between the Se optimum and hazardous concentrations that occurred in the body ,28,29,30occurred . A higheGóralska  conducteus above ,75,76,77ectively . DeForesectively  suggeste−1 [−1 was recorded [−1 [−1 [−1 [−1 [cutthroat trout (Oncorhynchus clarki lewisi) fry when the concentration of Se in the eggs was 20.6 µg g−1 (dw). Our study observed singular cases (0.3%) of spine deformities at hatch from groups with additional selenium dosage. Whereas in areas polluted with selenium, spine malformation is often noticed [In wild fish from different world areas, Se concentration in the body is typically below 1 µg g−1 ,73,78,79recorded . Se concrded [−1 . In contrded [−1 ,37,58,81 [−1 [−1 . Other a [−1 [−1  or 17 µg [−1 [−1 . Rudolph [−1 [−1  did not  noticed ,28,29,30 noticed ,83, part noticed ,85.−1 as SeO2 is recommended as optimal for practical application for studied sea trout.In selenium-poor areas, immersion of fertilised eggs in water enriched with Se during egg swelling can constitute a method to supplement the element to non-feeding stages of fish. The innovative method can be implemented in aquaculture to improve the growth of progeny and possibly fish survival in nature when larger individuals are stocked into watercourses. The selenium applied also had a slightly positive effect  on reducing egg mortality and increasing the hatching rate. Se concentration added to water should be adjusted to the selenium compound, fish species, and Se’s content in the yolk. Se concentration of 4–8 mg L


Sassafras tzumu were selected for spectral collection and ANTH, FLAV and NBI concentrations measurement by standard analytical methods. Partial least squares regression (PLSR), five spectra pre-processing methods, and four variable selection algorisms were conducted for the optimal model selection. Each trait model was simulated 200 times for error estimation.Plant traits related to nutrition have an influential role in tree growth, tree production and nutrient cycling. Therefore, the breeding program should consider the genetics of the traits. However, the measurement methods could seriously affect the progress of breeding selection program. In this study, we tested the ability of spectroscopy to quantify the specific leaf nutrition traits including anthocyanins (ANTH), flavonoids (FLAV) and nitrogen balance index (NBI), and estimated the genetic variation of these leaf traits based on the spectroscopic predicted data. Fresh leaves of 2 of 0.72 and mean RMSE of 0.10%, followed by FLAV and NBI model . High heritability was found for ANTH, FLAV and NBI with h2 of 0.78, 0.58 and 0.61 respectively. It shows that it is beneficial and possible for breeding selection to the improvement of leaf nutrition traits.The standard normal variate (SNV) to the ANTH model and 1st derivatives to the FLAV and NBI models, combined with significant Multivariate Correlation (sMC) algorithm variable selection are finally regarded as the best performance models. The ANTH model produced the highest accuracy of prediction with a mean RSpectroscopy can successfully characterize the leaf nutrition traits in living tree leaves and the ability to simultaneous multiple plant traits provides a promising and high-throughput tool for the quick analysis of large size samples and serves for genetic breeding program. Nitrogen (N) is one of the most essential nutrients in plant growth, which is needed for the improvement of grain yield and quality . ExcessiAnthocyanins (ANTH) are a group of water soluble flavonoid pigments that occur in all plant tissues. Anthocyanins are mostly related to a wide range of plant colour but often appear as red . In addiHowever, research on plant growth and the variation of N storage and remobilization have typically required labour intensive methods to measure the N concentration and index properties , such as atomic absorption spectrometry , 32, chrAlternatively, Near-infrared spectroscopy (NIRS) is a rapid, high-throughput technique that has been used for chemical components analysis in many fields. NIRS is a promising and reliable method that can be used for the assessment of a large number of samples , 46, 67.Ginkgo biloba leaf and four Indonesian herbal plant species, including Syzygium oleana, Piper betle, Jasminum and Graptophyllum pictum with NIR reflectance spectroscopy. NIR is a promising tool for tree breeding selection programs due to its robustness and capacity to screen large numbers of samples [Multivariate methods such as partial least squares regression (PLSR)  has been samples , 41.The robustness and reliability of model accuracy are largely determined by the spectra quality and feature selection. The combinations vibrations information and noise of the raw NIR spectra  will resThe joint analyses of chemometric statistics and variable selection algorithms has recently been used to eliminate the irrelevant variables and improve the model accuracy , 47. TheSassafras tzumu is a deciduous tree species that has colourful leaves in autumn. Zhejiang province in China is vigorously promoting the cultivation of colourful species making S. tzumu a famous tree species. It has been widely planted in Zhejiang province to develop the urban and mountain landscape [andscape .S. tzumu.Therefore, the aims of this research are to (1) test the capacity of reflectance spectroscopy to characterize the NBI, ANTH and FLAV with PLSR model; (2) find out the most optimal pre-processing method for these three leaf traits. (3) Identify the most important wavelength that related to NBI, ANTH and FLAV by four variable selection methods, including significant multivariate correlation (sMC), regularized variable elimination procedure (Rep), iterative predictor weighting (Ipw), and Genetic algorithm (Ga) variable selection; (4) estimate genetic parameters and correlations of NBI, ANTH and FLAV in S. tzumu were selected for our study from 6 different regions. Trees were planted in 2016 using a randomised complete block by a 2 m × 3 m spacing in Changle Forest Farm Nursery , Hangzhou, Zhejiang, China. Each family replicated 30 times with 5 replications and 6 individual trees per replication. In total, 1500 trees were planted.50 half-sib families of Samples spectra data was collected through 5–6 leaves of each tree from the top to bottom with similar color on the same side in October 2018. The NIR spectra data was taken from the upside surface of the leaves three times with a handheld fibre optic contact probe from a field-based spectrometer . Each spectrum took on average 32 scans with a range of 1100 to 2500 nm by a 6 nm resolution. All spectra were obtained from the leaves of 1500 trees, 500 trees leaves from these 1500 trees were sampled and placed in a marked paper bag and transferred to the refrigerator immediately for chemical measurement.Each leaf was ground into powder and being mixed with methanol for 24 h. 0.5 ml (1 mg/ml) extract of each sample was taken to mixed with methanol (1.5 ml), 10% aluminium chloride (0.1 ml), 1 M potassium acetate (0.1 ml) and distilled water (2.8 ml). The mixture was being placed under room temperature for 30 min and then measured at 415 nm for the absorbance by UV–Visible spectrophotometer . The flavonoid content of the sample was accessed by the value of absorbance density .A weighed circular piece cutting from each leaf was place into a mortar by a pestle ground with 100% methanol until the colour changed into white. The extract was being centrifuged for 6 min by 14,000 rpm at 4 °C and subsequently assayed by a UV–Visible spectrophotometer . It conducted the equation and specific absorption in the wavelength which was reported by Wellburn . The sol2) and root-mean-square error (RMSE) in each model derived from both calibration  and validation  were applied for the evaluation model performance.The original five different types of pre-processing spectra  combined with PLSR  algorithThe estimation of genetic parameters were measured by a multivariate restricted maximum likelihood (REML) linear mixed model, details can be found in Li et al. . The narwhere pls package [plsVarSel [prospectr package [lme4 package [ggplot2 package [R software (version 3.1.2)  was used package  in R waslsVarSel  for vari package  for NIR  package  for visu2 and RMSE for these three models in calibration  sets are 0.54 (range: 0.43–0.63), 0.47 (range: 0.35–0.58) and 0.36 (range: 0.26–0.45), in validation  sets are 0.54 (range: 0.28–0.75), 0.47 (range: 0.28–0.69) and 0.38 (range: 0.25–0.64) respectively. As for all spectral pre-processing models, SNV + 2nd derivative prediction model is found to be the highest well-performing for predicting ANTH concentration than the other pre-processing methods, with a mean R2Cal and RMSECal of 0.59 (range: 0.55–0.63), 0.11% (range: 0.11–0.12%), a mean R2Val and RMSEVal of 0.57 (range: 0.38–0.72), 0.11% (range: 0.09–0.13%), followed by 2nd, SNV + 1st, 1st, original with the mean of R2 in Cal is 0.56 (range: 0.42–0.75), 0.56 (range: 0.51–0.60), 0.53 (range: 0.48–0.59), 0.52 (range: 0.47–0.56), and RMSE 0.11% (range: 0.11–0.12%), 0.11% (range: 0.11–0.12%), 0.11% (range: 0.11–0.12%), 0.12% (range: 0.11–0.12), and in Val is 0.57 (range: 0.42–0.75), 0.54 (range: 0.30–0.70), 0.53 (range: 0.36–0.67), 0.52 (range: 0.32–0.72), and RMSE 0.11% (range: 0.09–0.13%), 0.12% (range: 0.10–0.14%), 0.12% (range: 0.10–0.14%), 0.11% (range: 0.09–0.14%) respectively. SNV shows the worst effect with the mean of R2 and RMSE for Cal and Val 0.49 (range: 0.44–0.54), 0.49 (range: 0.28–0.63), and 0.13% (range: 0.13–0.14%), 0.13% (range: 0.12–0.16%) respectively. However, 1st yields the best PLSR model in the prediction of FLAV and NBI than the other pre-processing model, with high mean R2Cal R2Val of 0.51 (range: 0.46–0.58), 0.52 (range: 0.29–0.68), and low mean of RMSECal, RMSEVal of 0.12% (range: 0.11–0.13%), 0.12 (range: 0.10–0.12%) in FLAV model and high mean R2Cal, R2Val of 0.39 (range: 0.33–0.45), 0.41 (range: 0.26–0.60), and low mean of RMSECal, RMSEVal of 0.05% (range: 0.05–0.05%), 0.05 (range: 0.04–0.06%) in NBI model respectively. The effect of SNV shows a poor prediction in the FLAV and NBI as well. The mean of R2Val is 0.40 (range: 0.26–0.64) and 0.47 (range: 0.29–0.64) respectively.Figure The relationship between the predicted and measured content of Cal and Val datasets by ANTH model with SNV + 2nd derivative spectra, FLAV and NBI model with 1st derivative spectra were plotted in Fig. The residual of the best processing spectra model for each leaf trait shows that all of these three models tend to be underpredicted when the measurement value is small. With the rise of the measurement value, the prediction value has the tendency of overprediction. The residual value of ANTH, FLAV and NBI model is between an acceptable range from − 0.3 to 0.3 , 0.58 (ranged from: 0.54 to 0.62), 0.44 (ranged from: 0.26 to 0.67), and the mean RMSEVal of 0.10% (range: 0.09–0.10%), 0.11% (range: 0.10–0.12%), 0.04% (range: 0.04–0.05%) respectively. The sMC_PLSR models reached a more stable prediction with less than 16% of full length of spectra on each leaf trait , which was lower than the result reported for fresh Ginkgo biloba leaf in different colors (R2CAL = 0.82 and RMSE = 2.62%) [The internal N cycling in trees  is a hot= 2.62%) . The var2Val of 0.72 (range: 0.69–0.75) and a mean of RMSEVal of 0.09% (range: 0.09–0.10%). Similar result was discovered in wine grapes by NIR hyperspectral imaging and PLSR model, which gave R2 of 0.84 and RMSEP of 0.013% for estimating ANTH content.Conversely, our prediction of ANTH content result illustrates a suitable accuracy than the other two leaf traits, with a mean RA robust statistical methodology for model calibration, which was first conducted by Couture et al.  to prediEucalyptus bosistoana trees, and Li et al. [The NIR spectra involves not only the favourable information but noise and irrelevant information which will encumbrance the accuracy of prediction model. Therefore, variable selection is regarded as an efficient way to find out the most important wavelengths which contributes the minimum error for model calibration and helps to reduce the model processing time for spectral models. Variables in the spectrum play a key role in the predictive accuracy of the model. The spectral information is extensive along with the relevant and irrelevant information, both of which will overlap to interfere the model construction of the useful information and the PLSR model with a specific trait . Thus, ii et al.  who founi et al.  is a posi et al. .. [Several important variables which are related to the ANTH, FLAV, and NBI have been selected similarly in each model, including the range at 2060, 2180, 2270, 2330 and 2440 nm for ANTH, 1070, 1235, 1950, 2220 nm for FLAV, and 1100, 1220, 1465, 1950, 2220 nm for NBI respectively. As reported by Ramirez et al. , the reg. , who strTrees N internal cycling is considered as one of the major ecology factors for tree growth and is an augment for the tree uptake of soil N . In addih2 ranging from 0.61 to 0.78. The leaf ANTH heritability of 0.78 in our study is similar to the result found by Yihu et al. [Populus tremula L.) [h2 = 0.58). It indicates that genetic control capacity is different between species even the same traits. Our study proved that there is also a potential for the selection of NBI traits in breeding programs even with less study on the estimation of NBI heritability.In this study, we continue to use the coefficients of 1/2.5 for the calculation of heritability of ANTH, FLAV and NBI traits based on our previous study to avoid the assembling of half-siblings and inbreeding effects . The modu et al.  who figumula L.) . For FLAmula L.)  which waThe consistence of families ranking of ANTH, FLAV and NBI indicates that the selection for a good leaf nutrition tree is workable, and the selection of qualified nutrition plant is supposed to involve multiple traits, which will afford a stable inheritance.In conclusion, NIR spectroscopy is potentially taken to estimate the nutrition related traits by fresh leaf. With the small prediction error, the tree breeding programs can be successfully achieved based on the relative prediction value. Our study provides an alternative way for the N index traits and open a door to the efficient analysis of the internal N cycling in trees. The pre-processing method and variable selection highly influence the performance of model prediction. Our study found that by using of 1st and SNV + 2nd derivative spectra processing method and sMC variable selection algorithm, the PLSR models have been highly improved. In addition, the repeated spectral statistical methodology that we applied provided an efficient way to deal with variation in calibration data and generate information on the response of plant nutrition traits with NIR spectra. NIR model serves as an efficient tool for the estimation of genetic parameters and breeding selection in high throughput way to improve the leaf traits quality.


This pilot survey aims to study the oral manifestations associated with COVID‐19 infection and report the prevalence of oral signs and symptoms in COVID‐19 patients.From May 15 to June 10, 2020, we used an online questionnaire containing the oral manifestations that are expected to be associated with the COVID‐19 infection. Adults in our survey who have been diagnosed with COVID‐19 positive were confirmed with reverse transcriptase PCR (RT‐PCR), and isolated in various hospitals in Cairo, Egypt.n = 19) did not have any symptoms associated with the oral cavity. The highest prevalence symptoms were dry mouth 39.7% (n = 23), gustatory dysfunction as 34.5% (n = 20) loss of salt sensation, 29.3% (n = 17) loss of sweet sensation, and 25.9% (n = 15) altered food taste, while the least prevalent symptoms were tongue redness 8.8% (n = 5), and gingival bleeding 7% (n = 4). The most frequently associated symptoms were loss of salt and sweetness, as reported by 27.6% of the participants. However, there was no significant association between the incidence of oral symptoms and demographic data  of the patients (p > 0.05).This pilot study included 58  COVID‐19 patients ages 18–46 years, and 13 (22.4%) were healthcare workers. Our results showed that 67.2% of the patients had at least one manifestation related to the oral cavity and salivary glands, and 32.8% (Based on limited data, COVID‐19 significantly impacts the oral cavity and salivary glands, as salivary gland‐related symptoms and taste disorders are highly prevalent in COVID‐19 patients. The protocol was registered in ClinicalTrials.gov (NCT04391881) and approved by the Research Ethics Committee, Faculty of Dentistry, Cairo University, Egypt. After reviewing the literature about the pathogenesis of COVID‐19 and its related manifestations, we developed an online Arabic survey using Google form to target adults (over 18 years old) who live in Cairo, Egypt, via Facebook advertisement Table .Since it was not feasible to examine the patients clinically, and to avoid selection biases due to convenience sampling, all participants were volunteers to minimize the risk of bias. We included only adults diagnosed with COVID‐19 positive, confirmed with reverse transcriptase PCR (RT‐PCR), and are isolated in various hospitals in Cairo, Egypt, from May 15 to June 10, 2020.We used a Google Form to create an online questionnaire containing multiple manifestations that were to be associated with COVID‐19 based on the available evidence. All symptoms included in this questionnaire were in four primary categories; the first category was the gustatory disorders described as a loss of salt sensation, loss of sweetness, and altered food taste. The second category was the symptoms associated with salivary glands infection described as dry mouth , difficulty swallowing, pain or swelling in the submandibular gland area, and pain or swelling in the parotid gland area. The third category was the oral mucosal changes  described as oral ulcers, spots of mouth or lips, tongue redness, gingival bleeding, and burning sensation. The last category was the patients with no symptoms related to the oral cavity and salivary glands.2.1p ≤ 0.05 within all tests, and p‐values were corrected for multiple comparisons using Bonferroni correction. Statistical analysis were conducted with Windows version 4.0.3 of the R software  COVID‐19 patients ages 18–46 years were included in this pilot study, while 13 (22.4%) were healthcare workers, and there were no significant differences in age or gender between participants (n = 19) did not have any symptoms related to the oral cavity. The high prevalence symptom was dry mouth 39.7% (n = 23), followed by gustatory dysfunction as 34.5% (n = 20) loss of salt sensation, 29.3% (n = 17) loss of sweet sensation, and 25.9% (n = 15) altered food taste. The prevalence of symptoms associated with salivary glands infection was 22.4% (n = 13) difficult swelling, 13.8% (n = 8) pain or swellings in the salivary gland or cheek, and 10.3% (n = 6) pain or swelling below mandible. While the oral mucosal changes were less prevalent as 22.4% (n = 13) of burning mouth sensation, 17.2% (n = 10) of ulceration, 13.8% (n = 8) of mouth and lip spots, 8.8% (n = 5) of tongue redness, and 7% (n = 4) of gingival bleeding  participants, while the dry mouth and gustatory impairment symptoms were often associated in 14 (27.6%) participants for each manifestation. However, the association between the tongue's redness either with lips or mouth spots, discomfort or swelling below the mandible, or gingival bleeding, was the least frequent, which occurred in one case  of the patients  occurs due to the viral infection to the olfactory cranial nerve or from rhinitis and nasal obstruction. The loss of smell (anosmia) is related to retro‐nasal olfaction which is a sensory process, combining ortho‐nasal smell and taste patterns and enabling people to perceive flavor. This mechanism is temporarily disrupted during upper respiratory infections due to mucosal inflammation and obstruction of the nasal passages, thereby physically blocking flavor and odor molecules from entering the olfactory cleft  could be due to several reasons . Evidence show that viral infection could weaken the immune system thus, creating secondary infections such as oral candidiasis. Candidiasis is the most prevalent opportunistic infection in HIV Nokta,  and is aConcerning the oral ulceration (17.2%), mouth and lip spots (13.8%), their prevalence was higher than other oral changes which supports the case report of ulceration and skin rash in COVID‐19 patients , which might have a misleading impact on the results. Despite the limitations, this pilot study's findings could help understand the oral manifestations associated with COVID‐19.5Our study indicates a significant impact of COVID‐19 on the oral cavity based on limited sample size. Salivary gland‐related symptoms and taste disorders are high prevalence in COVID‐19 patients, while the detailed oral manifestations associated with COVID‐19 and their pathophysiology required further investigation. Further clinical studies with larger sample sizes and detailed patient's history, are required to confirm our results and clarify the full impact of COVID‐19 on the oral cavity and salivary glands.All authors declare no conflicts of interest.Conceptualization: D.M.E.; Methodology: D.M.E., E.A.G., and A.G.A.K.; Data collection: W.S.A., R.H.A., and R.A.H.; Data analysis: A.G.A.K., W.S.A.; Writing original draft preparation: A.G.A.K., E.A.G., D.M.E., R.H.A., and R.A.H.; Writing–review and editing: A.G.A.K., D.M.E., and E.A.G.; All authors have read and agreed to the published version of the manuscript.Supplementary Table S1 An example of the online survey that we usedClick here for additional data file.Appendix S1: Supporting InformationClick here for additional data file.


Oryza sativa L.). Herein, some experiments were conducted so as to assess the effects of seed priming with selenium (Se) on the germination and growth of rice under hypoxia. The experiment was arranged in a completely randomized factorial design with two factors and five replicates. Factors included Se concentration  and duration of flooding stress . The experimental results showed that Se accelerated seed germination and increased emergence index and final emergence percentage. Additionally, Se increased shoot and root lengths and dry weights, but high Se concentration (60 μmol/L) reduced 18‐day‐old seedling dry weight under long‐term flooding (8 days). Furthermore, Se reduced malondialdehyde content and increased starch hydrolysis efficiency in seeds, superoxide dismutase, peroxidase, catalase, and glutathione peroxidase activities and seedling soluble protein and total chlorophyll contents. Se improved seedling total Se and organic Se contents while increasing total dry weight and yield. Notably, the highest yield was obtained after a 4‐day flooding period. Although Se priming favored rice seedling emergence and growth under flooding conditions, Se concentrations equal or above 60 μmol/L increased the risk of seedling death during long‐term flooding (≥8 days).Prevalent irregular rainfall, flooding for weed control, and unleveled fields in the middle and lower reaches of the Yangtze River all contribute to flooding stress on germination and growth of direct‐seeded rice ( Although rice is the only cereal crop that can germinate and extend its coleoptile under oxygen‐limited conditions  is an essential trace element for maintaining the normal functioning of many physiological processes , 2 (FD2), 4 (FD4), or 8 d (FD8) after rice sowing. Se levels comprised dry rice seeds primed by immersion in a sodium selenite (Na2O3Se) solution at Se 0 (Se0), 30 (Se30), or 60  μmol/L (Se60).The experiment was laid out in a split‐plot design with flooding duration (FD) as the main plot and Se concentration as the subplot. FD at four levels and Se at three levels were tested with five replicates, and 60 subplots were established in pots (Experiment 1) and in the field (Experiment 2). The FD levels comprised flooding with 10  cm  were added to the soil and mixed thoroughly. The mixed soil (5 kg) was packed into plastic pot with a diameter of 25 cm in diameter and a height of 30 cm. One hundred seeds were evenly sprinkled on the soil surface in each treatment, then immediately irrigated water by 10 cm, and maintained at a constant water level during inundation. After flooding duration ended, all treatments maintained a 1 cm water level until harvest (18 days). The FD were randomly arranged on a frame inside a glasshouse at 28/25°C day/night and a 16 h/day photoperiod with natural sunlight supplemented with sodium vapor lamps to maintain a light intensity > 350 μmol m−2 s−1.The soil used in Experiment 1 was a silty clay loam with 24% sand (2.00–.02 mm), 40% silt (.02–.002 mm), and 36% clay (<.002 mm), collected from the plow layer (0–20 cm) of an arable field in Yangtze University, Hubei Province, Southern China. Each kilogram of soil at pH 5.94 contained 34.52 g of organic matter, 224.19 mg of available N, 1.37 mg of available P, 127.24 mg of available K, and .29 mg of total Se. The soil was air‐dried, sieved to <5 mm, and homogeneous. Basal fertilizers 2, (NH4)2HPO4, and KCl, respectively. Specifically, 60% of N was applied as a basal fertilizer and the remaining 40% as a tillering fertilizer, whereas 100% of P2O5 and K2O were applied as base fertilizers. Treatments were arranged in a randomized complete block design with five replications and a plot area of 24.0 m2 (6 × 2 m). Manual weeding was performed in the early tiller, late tiller, and heading stages. Pest and disease incidence were intensively controlled.Experiment 2 was conducted in 2018–2019 at the Yangtze University farm in Jingzhou County, Hubei Province, China . The soil was the same as that used in Experiment 1. After land preparation, seeds treated in the same manner as those in Experiment 1 were sown directly in the field at a rate of 60 kg ha2.150), mean emergence time (MET), emergence index (EI), and final emergence percentage (FEP) of seeds were calculated as follows .n is the number of seeds emerged on day D and D is the number of days counted from the beginning of emergence, D  ≤  10.Et is the number of seeds emerged on day t and Dt is time corresponding to Et, 1  ≤  t  ≤  10.In Experiment 1, seedling emergence was counted daily using the method prescribed by the Association of Official Seed Analysts until the 10th day. A seedling was scored as ‘emerged’ when its hypocotyl length was ≥2  mm. The time to start emergence (TSE) of seeds were recorded. The time taken to 50% emergence  content using the thiobarbituric acid method  and polished rice (Experiment 2) were determined according to the method of Deng et al. .2.6p < .05.All experimental data are expressed as means ± standard errors (SE) of five replicates. The dates were subjected to the two‐way analysis (ANOVA) to determine the effects of flooding duration, Se treatments, and interaction between them, respectively. Significant differences between flooding duration and Se treatments among the same year were tested by Duncan's multiple range tests. The significance level was 33.150 was highest at FD4 and FEP was highest at FD2. Additionally, as Se concentration increased, TSE, E50, and MET decreased, whereas EI and FEP increased. It is worth noting that when flooding duration was less than 8  days, EI and FEP increased with the rising Se concentration, but when it extended for 8  days, EI and FEP in Se60 were lower than those in Se30. The results above indicate that long‐term flooding limits seed germination and Se priming promoted seed germination; however, when flooding duration was 8  days, Se60 had a negative effect on seed germination.Flooding duration, Se treatment, and their interaction significantly affected rice emergence, although the interaction had no significant effect on TSE Table . General3.20, shoot length, root length, and dry biomass in FD8 declined by 41.74%, 81.70%, and 79.21%, respectively, on the 10th day and by 35.90%, 48.06%, and 59.89%, respectively, on the 18th day. Shoot and root lengths and dry biomass increased with increasing Se concentration. Compared with Se0, shoot and root lengths and dry biomass of Se60 decreased by 37.36%, 18.18%, and 27.40%, respectively, on the 10th day and by 24.39%, 15.58%, and 27.41%, respectively, on the 18th day. Notably, Se60 significantly reduced the dry biomass of 18‐day‐old seedlings in FD8. These results indicate that flooding restricted seedling growth, whereas Se priming promoted seedling growth; however, Se60 reduced the dry biomass of 18‐day‐old seedlings in FD8.Flooding duration, Se treatment, and their interaction significantly affected seedling morphology, although the interaction had no significant effect on shoot length on the 18th day Table . As floo3.360 were 25.11% and 12.88% lower, respectively; however, α‐amylase activity and soluble sugar content in Se60 were 19.52% and 6.69% higher, respectively, compared with the Se0 treatment. These results indicate that Se priming reduced MDA content and accelerated starch hydrolysis.Se treatments significantly affected the biochemical attributes of rice seeds Table . MDA and3.44 and lowest in FD0. Furthermore, SOD and POD activities in FD0 were 14.63% and 26.15% lower, respectively, compared with FD4. In turn, CAT and GPx activities were highest in FD2 and lowest in FD8, whereas SOD and POD activities in FD8 were 25.74% and 19.20% lower, respectively, compared with FD2. Additionally, soluble protein and total chlorophyll contents decreased with increasing flooding duration. Thus, compared with FD0, soluble protein and total chlorophyll contents in FD8 increased by 35.51% and 44.72%, respectively. In contrast, SOD, POX, CAT, and GPx activities and soluble protein and total chlorophyll contents increased with increasing Se concentration. Thus, compared with Se0, these six physiological indicators increased by 55.53%, 129.00%, 225.68%, 9.72%, 75.86%, and 46.02%, respectively, in Se60. Se priming tended to increase antioxidant enzyme activities, as well as soluble protein and total chlorophyll contents, although the extent of such increase was different under different flooding duration treatments. In summary, short‐term flooding stimulated antioxidant enzyme activities. Conversely, long‐term flooding damaged the physiology of seedlings, but Se restored and in fact enhanced the physiology of seedlings under conditions of 0–8  days of flooding.Flooding duration, Se concentration, and their interaction significantly affected seedling biochemical attributes, although the interaction had no significant effect on GPx activity Table . Antioxi3.58, respectively, compared with FD0. Conversely, total Se and organic Se contents increased with increasing Se concentration; indeed, they increased by 2.407 and 1.454  mg  kg−1, respectively, in Se60, compared with Se0. Se content of polished rice ranged between .101 and .121  mg  kg−1 across treatments. These results indicate that flooding reduced the transport of Se from the seed to the growing seedling, whereas Se priming significantly increased Se content in seedlings.Flooding duration and Se treatments considerably affected seedling total Se and organic Se contents, although the interaction between them significantly affected only organic Se content Table . Total S3.64 and lowest in FD0. As a result, mean dry weight and yield in FD0 were 9.60% and 6.80% lower, respectively, compared with FD4. Both total dry weight and yield increased with increasing Se concentration. The corresponding mean values increased by 6.96% and 7.04%, respectively, in Se60, compared with Se0. However, the Se60 treatment group showed reduced total dry weight and yield in FD8 in both years. In summary, the highest yield was obtained under the 4‐day flooding treatment with Se priming.Flooding duration, Se concentration, and their interaction significantly affected total dry weight and yield in both years, but the interaction had no significant effect on Se content in polished rice in 2018 Table . In 2018460 in FD8, Se seed priming accelerated seed germination, increased EI and FEP, and promoted seedling growth  concentration and flooding duration on rice emergence. FD: flooding durationTable S1‐2. Effects of Selenium (Se) concentration and flooding duration on rice emergence. FD: flooding duration. TSE: time to start emergence, E50: time taken to 50% emergence, MET: mean emergence time, EI: emergence index, FEP: final emergence percentage. Means (n = 5) with different letters differ significantly at the 5% probability level based on Tukey's test.Table S2‐1. Analysis of variance for effects of Selenium (Se) concentration and flooding duration on rice morphology. FD: flooding durationTable S2‐2. Effects of Selenium (Se) concentration and flooding duration on rice morphology. FD: flooding duration. Means (n = 5) with different letters differ significantly at the 5% probability level based on Tukey's test.Table S3‐1. Analysis of variance for effects of Selenium (Se) concentration on biochemical attributes of rice seeds.Table S3‐2. Effects of Selenium (Se) concentration on biochemical attributes of rice seeds. One unit of the enzyme's activity is the amount of enzyme which released 1 μmol of maltose by 1 mL original enzyme solution in 1 min. Means (n = 5) with different letters differ significantly at the 5% probability level based on Tukey's test.Table S4‐1. Analysis of variance for effects of Selenium (Se) concentration and flooding duration on biochemical attributes of rice seedlings. FD: flooding durationTable S4‐2. Effects of Selenium (Se) concentration and flooding duration on biochemical attributes of rice seedlings. FD: flooding duration. SOD: superoxide dismutase, POD: peroxidase, CAT: catalase, GPx: glutathione peroxidase. Means (n = 5) with different letters differ significantly at the 5% probability level based on Tukey's test.Table S5‐1. Analysis of variance for effects of Selenium (Se) concentration and flooding duration on rice seedlings Se concentration. FD: flooding durationTable S5‐2. Effects of Selenium (Se) concentration and flooding duration on rice seedlings Se concentration. FD: flooding duration. Means (n = 5) with different letters differ significantly at the 5% probability level based on Tukey's test.Table S6‐1. Analysis of variance for effects of Selenium (Se) concentration and flooding duration on total dry weight, yield and Se content in polish rice in 2018–2019. FD: flooding durationTable S6‐2. Effects of Selenium (Se) concentration and flooding duration on total dry weight, yield and Se content in polish rice in 2018–2019. FD: flooding duration. Means (n = 5) with different letters differ significantly at the 5% probability level based on Tukey's test.Click here for additional data file.


To explore the clinical effects of sodium valproate and levetiracetam in the treatment of women with epilepsy during pregnancy. n = 56) and the levetiracetam group . The effects and the maternal and infant outcomes after treatment were compared between the two groups. The neuron-specific enolase (NSE), cognitive function-related parameters (brain-derived neurotrophic factor (BDNF) and myelin basic protein (MBP)), and related inflammatory factors (tumor necrosis factor- (TNF-) α and interleukin- (IL-) 6) levels were compared between the two groups before and after treatment.  The clinical data of 124 women with epilepsy during pregnancy who received monotherapy with antiepileptic drugs (AEDs) in our hospital from September 2017 to January 2020 were retrospectively analyzed. According to the type of medication taken by the patients, they were recorded as the sodium valproate group . After treatment, the probability of gestational hypertension, depression during pregnancy, low-weight infants, and neonatal deformities in the LEV group was lower than that in the VPA group (P < 0.05). After treatment, the levels of NSE, MBP, TNF-α, and IL-6 in the two groups decreased, and the levels of BDNF increased, and the LEV group changed significantly compared with the VPA group (P < 0.05).  After treatment, the total clinical effective rate of the LEV group was 91.18% higher than that of the VPA group 73.21%, and the frequency and duration of seizures were lower than those of the VPA group ( Compared with sodium valproate monotherapy, levetiracetam is more effective in controlling seizures and improving maternal and infant outcomes in women with epilepsy during pregnancy and can effectively regulate their neurological and cognitive functions and reduce the serum inflammation factor level. Epilepsy is a chronic transient brain dysfunction syndrome caused by the abnormally synchronized discharge of the patients' brain neurons . PatientSodium valproate is a traditional broad-spectrum AED, which is effective for the treatment of many types of epilepsy. Compared with the new AEDs of levetiracetam and oxcarbazepine, it has the advantage of low price. However, recent studies have shown that when it is used alone or in combination with other AEDs in patients with epilepsy during pregnancy, it has a higher probability of causing fetal malformations than other AEDs. Levetiracetam is a new type of AEDs. Compared with traditional AEDs and other new AEDs, it has a lower incidence of malformations associated with monotherapy in the fetus. Therefore, in recent years, its application in patients with epilepsy during pregnancy has become increasingly widespread. In this study, we observed the clinical effects and safety of sodium valproate and levetiracetam in the treatment of women with peripregnancy epilepsy. The aim was to explore the effects of the above two monotherapies on epilepsy control, maternal and infant outcomes, neurocognitive function, and body inflammatory level in peripregnancy women with epilepsy. The report is as follows.n = 56) and the levetiracetam group . There was no statistical difference between the general information in P > 0.05).This is a retrospective analysis of the clinical data of 124 women with epilepsy during pregnancy who received AEDs monotherapy in our hospital from September 2017 to January 2020. Case inclusion criteria: met the diagnostic criteria of the International Anti-Epilepsy League , 22–38 yBoth groups of patients received routine antiepileptic care and management during the treatment period, including reasonable diet arrangements, avoiding patients from eating foods that may induce epileptic seizures, and providing psychological counseling interventions for patients to relieve the psychological pressure of patients. On this basis, the VPA group received sodium valproate sustained-release tablets (Sanofi (Hangzhou) Pharmaceutical Co., Ltd., approval number: H20010595, specification: 0.5 g × 30 tablets) monotherapy. Usage and dosage: oral 30 min after meal, 2 times/d, initial dose is 10 mg/kg, increase to 20 mg/kg after 1 week, and maintain treatment. The LEV group received levetiracetam tablets  monotherapy. The usage and dosage are oral 30 min after meal, 2 times/d, the initial dose is 10 mg/kg, increased by 10 mg/kg every week, so that the patient's drug dose at late pregnancy is controlled at 1800 ± 200 mg/kg.Therapeutic effect: the criterion of efficacy was based on the revised scoring standard of the International Anti-Epilepsy League . Among tMaternal and infant outcomes: statistics and comparison of the number of cases of placenta previa, premature delivery, miscarriage, gestational hypertension, and depression during pregnancy between the two groups of patients and statistics and comparison of the two groups of fetal cases of fetal distress, stillbirth, low body mass, and neonatal deformities.α and interleukin- (IL-) 6) levels were compared between the two groups before and after treatment. Detection method: before and after treatment, 6 mL of venous blood was drawn in the morning, centrifuged at 2800 r/min for 8 min, and the supernatant was separated and placed in a refrigerator at −30°C for testing. The detection was carried out by the enzyme-linked immunosorbent method, and the kit was provided by Epson (Shanghai) Biotechnology Co., Ltd.Serological indicators: the neuron-specific enolase (NSE), cognitive function-related parameters (brain-derived neurotrophic factor (BDNF) and myelin basic protein (MBP)), and related inflammatory factors (tumor necrosis factor- (TNF-) s), and t-test analysis was used for comparison. The count data were expressed as (%), and χ2-test analysis was used for comparison. P < 0.05 indicated that the difference was statistically significant.Data analysis was processed by SPSS 22.0 software. The measurement data were expressed as (P < 0.05)  .P < 0.05)  times/month was less than that in the VPA group (2.41 ± 0.23) times/month, and the duration of seizures in the LEV group (2.84 ± 1.01) minutes/time was less than that in the VPA group (3.96 ± 1.21) minutes/time ( < 0.05) .P < 0.05) , 16.07% (9/56), 16.07% (9/56), 26.79% (15/56), and 28.57% (16/56), respectively; the incidence of placenta previa, premature delivery, miscarriage, gestational hypertension, and depression during pregnancy in the LEV group was 7.35% (5/68), 13.24% (9/68), 10.29% (7/68), 11.76% (8/68), and 11.76% (8/68), respectively. The incidence of fetal distress, stillbirth, low body mass, and neonatal deformities in the VPA group was 17.86% (10/56), 0.00% (0/56), 83.93% (47/56), and 26.79% (15/56), respectively; the incidence of fetal distress, stillbirth, low body mass, and neonatal deformities in the LEV group was 8.82% (6/68), 0.00% (0/68), 54.41% (37/68), and 5.88% (4/68), respectively. The probability of gestational hypertension, depression during pregnancy, low-weight infants, and neonatal deformities in the LEV group was lower than that in the VPA group ( < 0.05) .μg/mL, IL-6 levels were 13.12 ± 1.66 and 13.15 ± 1.61 pg/mL, and TNF-α levels were 3.39 ± 0.43 and 3.41 ± 0.43 ng/mL, respectively. After treatment, the NSE levels of the VPA group and LEV group were 9.13 ± 2.37 and 7.11 ± 2.25 ng/mL, BDNF levels were 30.24 ± 3.52 and 33.68 ± 3.65 pg/mL, MBP levels were 2.56 ± 0.23 and 1.98 ± 0.20 μg/mL, IL-6 levels were 9.42 ± 1.15 and 8.20 ± 1.06 pg/mL, and TNF-α levels were 1.96 ± 0.20 and 1.24 ± 0.14 ng/mL, respectively. After treatment, the levels of NSE, MBP, TNF-α, and IL-6 in the two groups decreased, and the levels of BDNF increased, and the LEV group changed significantly compared with the VPA group (P < 0.05)  .According to incomplete statistics, about 0.3–0.7% of women of childbearing age in the world have epilepsy during pregnancy and 0.3–0.5% of children come from women with epilepsy –15. In aγ-aminobutyric acid in the brain, inhibiting the abnormal discharge of neurons, preventing the influx of calcium ions, and maintaining the stability of neuronal cell membranes [Sodium valproate is the first-line AEDs commonly used in clinical practice. It has the advantages of broad antiepileptic spectrum, relatively cheap price, low concentration of drug required, and long action time. Its mechanism of action is related to the promotion of the high expression of the inhibitory neurotransmitter embranes . At the embranes . The advembranes . Therefoembranes . A studyembranes . In thisembranes . This maLevetiracetam is a pyrrolidone derivative, which can specifically bind to the central nerve synaptic vesicle protein 2A subunit to inhibit the release of neurotransmitters, block the abnormal discharge of neurons, and ultimately reduce seizures . In addiα can excite neurotransmitters, and elevated levels of IL-6 can change neuronal action potentials. The interaction between the two can excite neurons, cause hippocampal damage, and aggravate epilepsy [α, and IL-6 in the two groups decreased, and the levels of BDNF increased, and the LEV group changed significantly compared with the VPA group. It is suggested that compared with sodium valproate monotherapy, levetiracetam monotherapy is more effective in improving neurocognitive function and serum inflammation levels in women with epilepsy during pregnancy. This once again confirmed the good prospects of levetiracetam in the treatment of women with epilepsy during peripregnancy.Epilepsy is mostly caused by the abnormal discharge of brain neurons. Long-term repeated seizures can trigger events such as hippocampal neuron regeneration and other central nervous system damage, which in turn leads to cognitive decline in patients . In addiepilepsy . After tIn summary, compared with sodium valproate monotherapy, levetiracetam is more effective in controlling seizures and improving maternal and infant outcomes in women with epilepsy during pregnancy and can effectively regulate their neurological and cognitive functions and reduce the serum inflammation factor level.


This study aimed to explore the extent of influence of sex hormones on HbA1c levels in non-diabetic population.Sex hormones are strongly linked to the occurrence and development of diabetes, and influence glycated hemoglobin , total testosterone (TT), and sex hormone binding globulin (SHBG) were measured by chemiluminescent microparticle immunoassays.A total of 1409 non–diabetic subjects, including 601 men and 808 postmenopausal women were recruited from Shanghai community. HbA1c was 5.6 (5.4–5.9) % in all subjects, with 5.6 (5.4–5.8) % in men and 5.7 (5.5–5.9) % in postmenopausal women. After adjusting for age, body mass index (BMI), and hemoglobin, E2 was positively correlated with HbA1c in men , and SHBG was inversely correlated with HbA1c  in women. Other hormones were not correlated with HbA1c . Multivariate linear regression analysis showed that, except for traditional factors, such as age, hemoglobin, and BMI, E2 was another determinant of HbA1c  in men; besides, in women, SHBG was another determinant of HbA1c , except for age and systolic blood pressure.The level of HbA2 and SHBG could influence HbA1c levels in non-diabetic population.After controlling for confounding factors, two sex hormones, as EThe online version contains supplementary material available at 10.1186/s12902-022-01002-w. Sex hormones, including estrogen, progesterone, and androgens, are synthesized by the human gonads and adrenal cortex. Their main role is to promote development of the reproductive system and maintain secondary sexual characteristics. More than that, low circulating levels of testosterone and sex-hormone-binding globulin (SHBG) were reported to be associated with increased cardiovascular risk in men, possibly due to effects on insulin resistance and glycemia. Recent studies have found that men with low serum testosterone levels had increased risk of diabetes; however, it was in contrast to that in women –3. Addit1c has long been regarded as the gold standard for evaluating long-term blood glucose control in patients living with diabetes in clinical routines. In recent years, standardized detection of HbA1c has been constantly improved, leading to sufficient sensitivity and specificity for HbA1c as an indicator of diabetes diagnosis , and C-reactive protein (CRP). Additionally, all the non-diabetic subjects were tested for hemoglobin, and blood samples were collected after 75 g oral glucose tolerance test to determine the 2-h blood glucose (2hPG). All laboratory indicators were measured using the standard methods. The homeostasis model assessment of insulin resistance (HOMA-IR) was as follows: HOMA-IR = FINS (mU/L) × FPG (mmol/L)/22.5 ; Kt = 1 × 109 L/mol, N = Ka × Ca + 1, where Ka = 3.6 × 104 L/mol; and Ca is Alb level. The sensitivity of E2, TT, and SHBG estimation was < 2.5 pg/mL, < 0.1 ng/mL, and <  0.3 mmol/L, respectively. The intra-assay and inter-assay coefficients of variation for E2 were < 2.40% and < 2.70%, respectively; the intra-assay and inter-assay coefficients of variation for TT were < 2.16% and < 2.23%, respectively. The intra-assay and inter-assay coefficients of variation for SHBG were < 2.80% and < 4.30%, respectively.HbAt-test was used to compare two groups with normal distribution, whereas Wilcoxon rank sum test was used for skewed distribution between two groups. Partial correlation analysis was conducted to analyze the potential correlation between the levels of HbA1c and sex hormones. A multivariate linear regression was performed to examine this correlation in men and women, respectively. All P values were two-tailed, and P < .05 was considered statistically significant.The research data was analyzed using SPSS, version 20.0 . All variables were tested for normality. Normally distributed variables are presented as mean ± standard deviation, and non–normally distributed variables are presented as median and interquartile range. Student’s 1c level was 5.6 (5.4–5.9) % in all the subjects, with 5.6 (5.4–5.8) % in men and 5.7 (5.5–5.9) % in postmenopausal women. Men had a median E2 level of 99.1 (84.4–121.1) pmol/L, median TT of 20.12 (15.5–24.9) nmol/L, median BT of 12.5 (10.0–15.9) nmol/L, and median SHBG of 40.7 (30.4–52.8) nmol/L. Postmenopausal women had a median E2 level of 36.7 (18.4–47.7) pmol/L, median TT of 0.9 (0.8–1.2) nmol/L, median BT of is 0.3 (0.2–0.3) nmol/L, and median SHBG of 53.7 (38.8–72.8) nmol/L  ; whereas, women tended to have higher HbA1c, HOMA-IR, TC, HDL-c, and LDL-c levels than that in men .A total of 1409 cases with an average age of 60.4 ± 6.2 years were enrolled in this study, including 601 men, and 808 postmenopausal women with a median menopause duration of 10 (6–15) years. HbA2 with HbA1c in men and SHBG with HbA1c in women. Partial correlation analysis after adjusting for age, BMI, and hemoglobin levels showed that only E2 was positively correlated with HbA1c in men ; BT was marginally correlated with HbA1c . TT and SHBG were not correlated to HbA1c . In women, only SHBG was negatively correlated with HbA1c , while other hormones were not correlated with HbA1c   .A multivariate linear regression model was constructed with data from men and postmenopausal women to analyze the association of sex hormones with HbA1c, and was conducted in both men and postmenopausal women. We found that E2 was a positive factor for HbA1c in men while SHBG was a negative factor for HbA1c in postmenopausal women.To our knowledge, the present study was the first to assess the association between a relatively complete set of sex hormones and HbA1c levels [At a cellular level, testosterone increases the expression of insulin receptor β subunit, insulin receptor substrate-1, protein kinase B and glucose transporter type 4 in adipose tissue and adenosine 5′-monophosphate-activated protein kinase expression and activity in skeletal muscle , 17. Henc levels .1c in men with type 2 diabetes [1c levels in these individuals. We found that serum BT and HbA1c were marginally correlated in men; while after adjusting for other factors, BT was correlated with other sex hormones, but no correlation was found between BT and HbA1c. Additionally, we found that E2 was positively correlated with HbA1c in men. Mean age of men included in this study was 60.3 years old. Moreover, testosterone levels gradually decrease with age [Previous studies showed that serum BT was positively correlated with HbAdiabetes , but thidiabetes . Here, wwith age , while twith age , 23. Thu1c in postmenopausal women. Previous study showed a significantly negative correlation between SHBG and HbA1c in non-diabetic postmenopausal women (n = 200) after adjusting for age and BMI [1c. Additionally, we found that hemoglobin was a negative factor, while BMI was a positive factor for HbA1c in men. Age was a positive factor for HbA1c, in both men and postmenopausal women, consistent with results of previous studies [We also found that SHBG was negatively correlated with HbA and BMI . In this and BMI . Testost and BMI . The aff and BMI . This ma studies , 28.1c levels. Therefore, prospective studies with a larger sample size and different age groups are needed to validate these findings.This study has some limitations. First, the study population only included middle-aged and elderly individuals, and hence, the results may not be generalized to entire community. Second, the cross–sectional study design could not determine the causal relationship between changes in sex hormone and HbA1c levels in non-diabetic population. Moreover, levels of E2 were independently and positively correlated with HbA1c in men, and those of SHBG were independently and inversely correlated with HbA1c in postmenopausal women.In summary, sex hormones influence HbAAdditional file 1.


Background: The nanotechnologies have been applied for dental restorative materials manufacturing such as glass ionomer cement, composites, tooth regeneration, and endodontic sealers. The study aimed to investigate the chemical bond of conventional glass ionomer cement and to evaluate the addition of different concentrations of silver nanoparticles (AgNPs) on the quality of the chemical bond of glass ionomer cement to primary dentin.Methods: Silver nanoparticle (AgNP) powder was added in concentrations of 0.2, 0.4, and 0.6% to the conventional powder of GIC Fuji II. Then, the powder was added to the liquid and mixed with the recommended powder/liquid ratio of 3.6:1 g. The Fourier-transform infrared spectra (FTIR) of teeth with 0.2%, 0.4%, and 0.6% w/w of silver nanoparticles in GIC fills and the control tooth were obtained. The conventional glass ionomer was used as a control group. The control and the plain silver tooth were subjected to FTIR analysis using an ATR–FTIR spectrophotometer  with zinc selenide (ZnSe) ATR crystal  and OPUS v7.5 software. All spectra were recorded in the range of 500–3,500 cm−1 in the transmission mode with an ATR module.Results: The AgNPs added at 0.2, 0.4, and 0.6% concentration to GIC provided some information in the context of bond interaction with the dentin. Various bond peaks were seen for calcium, carbonate, phosphate, and amide. In our study, only the amide and phosphate were generated. The amide peaks were almost similar to the control, 0.2%, 0.4%, and 0.6%, with the peaks in the range of 1250–1650 cm−1. There was a clear shift in the phosphate peak from the control, 0.2, and 0.4%, which was about 1050 cm−1, whereas for 0.6%, there was a clear shift from 1050 cm−1 to 880 cm−1.Conclusion: GIC supplemented with AgNPs showed that a concentration above 0.4% of AgNPs altered the bond quality in dentin interaction. In conclusion, adding AgNPs at a minimal level improves the mechanical properties and maintains the same bond quality as GIC. The glass ionomer cement (GIC) was first discovered by Streptococcus mutans, beneath the restorations : 50 percent distilled water, 40 percent polyacrylic acid, and 10 percent polybasic carboxylic acid ] and silver nanoparticles (AgNPs) (<100 µm in size) from Sigma Aldrich .The proposal was registered with the research center of Riyadh Elm University (FPGRP/43835007/334), and ethical approval was obtained from the Institutional Review Board of the institution.in vitro study, a conventional GIC  (f) and silver nanoparticle powders (AgNPs) <100 nm particle size  (n = 10), GIC with 0.2% silver nanoparticles (AgNPs) (n = 10), GIC with 0.4% silver nanoparticles (AgNPs) (n = 10), and GIC with 0.6% AgNPs (n = 10) (In this  States)  were pur States) , and thr(n = 10) . The matThe extracted teeth were collected from an operating room and dental clinic. The teeth were carefully examined to ensure the absence of debris. For 1 month, the teeth were stored in a 0.1% thymol solution with 0.9% isotonic sodium chloride (5°C) until the beginning of the experiment. We used a diamond separating disc  at a slow-speed handpiece with continuous water cooling, perpendicular to the tooth’s long axis, and sectioned approximately 2.0 mm of the tissue along with the cusps without exposing the pulp . The 40 −1 in the transmission mode with an ATR module. The FTIR vibration range mode wavenumber was from 500 to 3500 cm−1. The FTIR analysis of GIC showed a similar interaction with the dentin compared to the GIC, with 0.2 and 0.4% AgNPs. These vibrational groups were part of the cross-linking reaction and aging time. In addition, the FTIR spectra showed the vibration of Ag in molecular water associated at the range of 3300 cm−1. The vibration band then shifted to 880 cm−1. This band was related to the bonding structure present in the GIC sample with 0.6%. The other band existing at ∼1,550 cm-1 referred to the formation of the asymmetric COOH band from the PAA.Fourier-transform infrared spectroscopy (FTIR)  providesOnly descriptive analysis was carried out, and statistical analysis was not performed in this study due to the qualitative characteristics of the data resulting from FTIR .−1, 1365 cm−1, 1456 cm−1, 1637 cm−1, and 1740 cm−1, respectively, were significant. The results of the bioactive evaluation of glass ionomer cement, carried out by Fourier-transform infrared spectroscopy, are shown in −1, 1,365 cm−1, 1,412 cm−1, 1490 cm−1, and 1556 cm−1, respectively, were significant. The GIC with silver nanoparticles (AgNPs) 0.2% had various peaks, of which v1, v2, v3, v4, and v5 with 1,048 cm−1, 1,368 cm−1, 1,410 cm−1, 1492 cm−1, and 1561 cm−1, respectively, were significant. The results of the bioactive evaluation of glass ionomer cement and silver nanoparticles (AgNPs) 0.2% with dentin, carried out by Fourier-transform infrared spectroscopy, are shown in The results of the bioactive evaluation of silver nanoparticle cement, carried out by Fourier-transform infrared spectroscopy, are shown in −1, 1,360 cm−1, 1,418 cm−1, 1,485 cm−1, and 1,548 cm−1, respectively, were significant. The GIC with silver nanoparticles (AgNPs) 0.6% had various peaks, of which v1, v2, v3, v4, and v5 with 8,080 cm−1, 1,365 cm−1, 1,408 cm−1, 1,493 cm−1, and 1,565 cm−1, respectively, were significant. The results of the bioactive evaluation of glass ionomer cement with silver nanoparticles (AgNPs) 0.6% with dentin, carried out by Fourier-transform infrared spectroscopy, are shown in −1, 1242 cm−1, 1546 cm−1, and 1655 cm−1, respectively, were significant.The results of the bioactive evaluation of glass ionomer cement with silver nanoparticles (AgNPs) 0.4% with dentin, carried out by Fourier-transform infrared spectroscopy, are shown in The present study has been carried out to investigate the effect of the bonding interaction of AgNP incorporation into GIC onto the dentin surface. Three concentrations  were added, with the control being the GIC. This study showed a greater variation of the transmission bands after the increased addition of silver nanoparticles, indicating a change in bond interaction with the dentin. Pure GIC has the disadvantage of less wear resistance, and it cannot withstand the masticatory forces. The common reason for the low resistance of GIC to fracture is the presence of voids in the cement matrix, which are formed by the inclusion of air during cement mixing. These voids may act as stress raisers and concentrators and eventually weaken the mechanical properties of the set cement . Many prRecent studies suggest that the voids tend to be filled with nanoparticles incorporated into GIC . In thisThe small sizes of the silver nanoparticles incorporated into GIC and the improved packing of particles within the matrix of the set cement may explain the improvement of the flexural and compressive strengths of the AgNP-containing GIC. Incorporating AgNPs into GIC may also result in a broader particle size distribution range. Therefore, these small silver nanoparticles can occupy the empty spaces between the larger glass particles and provide an additional bonding site for the polyacrylic polymer . ConsideThe present study aimed to examine the quality of the bonding interaction without changing the ideal bond quality achieved with GIC. However, the study focused on the exchanges that occurred when the cement was brought into contact with dentin. It is essential to differentiate between short-term and long-term interactions. Short-term interactions occur when the freshly prepared glass ionomer cement is brought into contact with dentin. They correspond to the rapid inter-diffusions between the dentin elements and the glass ionomer cement when the cement is not entirely set. These inter-diffusions enable the GIC to adhere to dentin. This ceases once the cement has been developed completely. Long-term interactions correspond to the slow diffusion of some elements of the glass ionomer cement through dentin. They can be caused by water in the buccal environment .−1, 1,631 cm−1, and 1,383 cm−1. Similar peaks were evident in the present study with little variation, showing various peaks, such as v1, v2, v3, v4, and v5 with 1,068 cm−1, 1365 cm−1, 1456 cm−1, 1637 cm−1, and 1740 cm−1, respectively. FTIR of the dentin surface showed several amide peaks  in the range between 1,250 and 1,650 cm−1, and the phosphate intensity ranged slightly over 1,000 cm−1 . However, FTIR has the advantage that IR spectra, with an acceptable signal/noise ratio, can be collected from areas measuring several hundred square micrometers in a matter of minutes.The statistical analysis was not carried out in the present study, based on the study and descriptive analysis carried out by In vitro studies already have good evidence, but we suggest in vivo studies to improve the quality of the restorations in a clinical condition.We warrant further research to examine the addition of other substances to GIC and their effect on the bond strength of this material. The descriptive analysis in the present study showed that any concentration beyond 0.4% of AgNPs altered the bond quality with dentin interaction. In conclusion, adding AgNPs to a minimum improves the mechanical properties and maintains the same bond quality as GIC.