ObjectiveGlucose and lactate levels in patients at the time of admission have been studied in diverse patient groups. Some studies suggest that elevated glucose levels at admission predict worse outcomes. Elevated Lactate levels have also been reported to be directly associated with increased mortality. We wanted to determine if the combination of admission glucose and lactate levels improves the predictability of inpatient mortality and length of stay (LOS).MethodsThis is a retrospective study. We included all adult patients admitted at an academic medical center from October 1, 2015 to September 30, 2016. We collected basic clinical information, including age, gender, admission glucose and lactate levels, LOS, and mortality. We separated outcomes based on glucose and lactate levels by dividing them into quartiles. We also stratified patients based on normal lactate (<2.0 mmol/L), high lactate (2.0-4.0 mmol/L), and very high lactate (>4 mmol/L) levels; and on normal glucose (60-140 mg/dl), high glucose (140-200 mg/dl), and very high glucose (>200 mg/dl) levels.ResultsA total of 5,436 adult patients were included in our study. The median age was 58 years, and 57% of the patients were male. The median LOS was 6 days, and the overall in-hospital mortality rate was 11%. When the patients were separated in quartiles based on admission glucose values, mortality was higher in the 4th quartile (≥173 mg/dL): 14.87%, probability value (p): <0.001. When the patients were separated in quartiles based on lactate levels, the mortality was higher in the 4th quartile (≥2.23 mmol/L): 21.95%, p: 0.001. When the patients were paired according to normal, high, or very high lactate and glucose levels, the groups that had higher mortality were as follows: normal glucose/very high lactate: 32.43%; high glucose/very high lactate: 34.04%; and very high glucose and very high lactate: 39.15%. The groups with very high glucose and very high lactate had increased odds of mortality when compared with the other groups (p: <0.001).ConclusionsAdmission glucose and lactate levels provide useful information in the estimation of inpatient mortality. The LOS was shortened in the groups with higher glucose, lactate, or both. The combination of glucose and lactate levels predicted mortality better than either value alone.  The identification of potential clinical indicators that predict patient outcomes is very important in current medical practice ,2. ThereThis is a single-center, retrospective study. It included all adults (≥18 years old) admitted to University Medical Center in Lubbock, Texas, from October 1, 2015 to September 30, 2016, whose admission glucose and lactate levels were recorded. Requests for laboratory tests were based on individual patient management decisions. Patients who were <18 years of age and >89 years of age were excluded. General demographic information was collected. This included age, gender, LOS, and mortality. University Medical Center is a 500-bed referral center in West Texas. The patients were stratified according to their lactate levels into three categories: lactate levels of <2 mmol/L; lactate levels of 2.0-4.0 mmol/L; and lactate levels >4.0 mmol/L. The respective LOS and mortality were calculated for each category. The patients were then classified according to their respective quartiles for their glucose and lactate levels, and their respective LOS and mortality were calculated. Patients were also classified according to their glucose/lactate molar ratios into quartiles, with their LOS and mortality for each quartile subsequently calculated.Finally, the patients were arbitrarily classified according to their paired glucose and lactate levels. We determined the following categories: normal lactate: <2 mmol/L; high lactate: 2.0-4.0 mmol/L; and very high lactate: >4 mmol/L. These were paired with glucose levels classified as follows: normal glucose: 60-140 mg/dL; high glucose: 140-200 mg/dL; and very high glucose: >200 mg/dL. The respective LOS and mortality were calculated for each group.Descriptive statistics were used to describe the characteristics of the study cohort. Categorical variables were summarized as frequencies, and continuous variables were summarized using medians and quartiles or percentiles, as appropriate. Univariate logistic regression was used to test if there is a significant association between mortality and assigned categories. Odds ratios (ORs) were calculated to evaluate these associations. Comparisons of LOS across categories were performed using the Kruskal-Wallis rank sum test. The statistically significant level was set at 0.05. Multiple testing adjustment was not performed. Analyses were performed using SAS software, Windows version 9.3 . A total of 5,436 patients were included in this study. The median age was 58 years (25th-75th percentile: 44-69 years); 57% of the patients were male. The median LOS was 6 days (25th-75th percentile: 3-11 days). The overall in-hospital mortality was 11.04%. The distribution of glucose, lactate, and glucose/lactate ratios for representative diagnoses are shown in Table The patients were stratified according to their lactate levels. Those with a lactate level of <2.0 mmol/L had a mortality rate of 7.18%; those with a lactate level between 2.0-4.0 mmol/L had a mortality of 13.16%; and those with a lactate level of >4 mmol/L had a mortality of 36.04%. Specifically, when treated as a continuous predictor, a one-unit increase in lactate was associated with a 32% increase in the odds of mortality (p: <0.001) : 9.44%; 2nd quartile (glucose: 102-125 mg/dl): 9.26%; 3rd quartile (glucose: 125-173 mg/dL); 10.53%; and 4th quartile (glucose: ≥173 mg/dl): 14.87%. Specifically, when comparing the patients in quartiles 1, 2 and 3, the patients in the 4th quartile had a 68%, 71%, and 49% increase in the odds of mortality, respectively (p: <0.001). The LOS was shorter for those in the 4th quartile (p: <0.001) : 5.56%; 2nd quartile (lactate: 1.05-1.45 mmol/L): 6.83%; 3rd quartile (lactate: 1.45-2.23 mmol/L): 9.72%; and 4th quartile (lactate: >2.23 mmol/L): 21.95%. Compared with patients in quartiles 1, 2 and 3, the patients in the 4th quartile had a 377%, 284%, and 161% increase in the odds of mortality, respectively (p: <0.001). The LOS was shorter for those in the 4th quartile (p: <0.001) : 19.72%; 2nd quartile (3.3-5.1): 9.35%; 3rd quartile (5.1-7.5): 8.98%; and 4th quartile (>7.5): 6.11%. When comparing the patients in the 1st quartile glucose/lactate molar ratio, the patients in quartiles 2, 3, and 4 had a 58%, 60%, and 74% decrease in the odds of mortality (p: <0.001) ; the highest mortality was in the groups with normal glucose/very high lactate 32.43%), high glucose/very high lactate (34.04%), and very high glucose/very high lactate levels (39.15%). In the normal glucose category, compared with patients in normal and high lactate groups, patients in the very high lactate group had a 603% and 212% increase in the odds of mortality (p: <0.001), respectively. In the high glucose category, compared with patients in the normal and high lactate groups, patients in the very high lactate group had a 345% and 295% increase in the odds of mortality (p: 0.001), respectively. In the very high glucose category, compared with patients in normal and high lactate groups, patients in the very high lactate group had a 897% and 290% increase in the odds of mortality (p: <0.001), respectively. In addition, patients in the very high glucose/very high lactate group had the shortest LOS   comparing glucose levels, lactate levels, and glucose/lactate ratios shows a higher AUROCs for lactate levels and glucose/lactate levels when compared to glucose levels alone, indicating better performance of lactate levels and glucose/lactate ratios for predictors for mortality Figure .In this study, we found that the combination of two well-known markers provides valuable information in the early identification of patients with worse outcomes. The group of patients with a combination of very high glucose and very high lactate had the highest mortality in the study cohort. Our results indicate that this combination provides better identification of the sickest patients when compared to either lactate or glucose levels alone.It has been well established that elevated lactate levels are associated with increased morbidity, LOS, and mortality ,5,6. HypThe relationship between increased glucose levels and patient outcomes has also been studied, and there is evidence supporting the association of high glucose values with increased morbidity, a longer length of hospitalization, increased readmission, and worse outcomes ,15,16,17These associations may be related to the metabolic changes caused by an acute illness in glucose and lactate metabolism ,21,22. IVery few studies have analyzed combined glucose and lactate levels as predictors of patient outcomes. Kaukonen et al. found that there is an independent association with hyperglycemia and hyperlactatemia and increased mortality. But the association of hyperglycemia disappeared when hyperlactatemia was considered in combination, suggesting that elevated lactate levels may have a stronger association with poor outcomes . Green eWe found a decreased LOS in the groups with the highest glucose, lactate, or both. The explanation for these results is uncertain. This is in contrast to previously published studies, where elevated lactate levels have been associated with increased LOS . One expSome of the strengths of this study include a large population sample, with patients included from multiple clinical settings and services. The authors recognize the limitations of a retrospective study, including the inability to discriminate stress-induced hyperglycemia from diabetes mellitus , the lack of serial measurements of lactate and glucose values, and the inability to determine the causes of death.The combined use of glucose and lactate levels provides useful information for early-risk stratification to identify those patients with worse clinical outcomes. Patients with very high glucose and very high lactate levels need more evaluation and intensive care.


The effects of pH, PAC coagulant dose alone and with polymers dose in various combinations was studied by jar tests. To compare the removal efficiencies of turbidity, total suspended solids (TSS), chemical oxygen demand (COD), and color at different levels, we run multivariate analysis of variance. Regarding the economic evaluation, we applied the incremental cost-effectiveness ratio. PAC had the best performance in pH 7 and in optimal dose of 400 mg/L; so that removal efficiency of wastewater turbidity, TSS, COD and color were 99.63%, 99.7%, 47.5% and 50.38%, respectively. The best removal efficiency for wastewater turbidity, TSS, COD and color were 99.87%, 99.89%, 87.5% and 93.02%, respectively which were obtained by combination of anionic polymer (1.5 mg/L) with PAC (300 mg/L). Furthermore, with combination of PAC + anionic + non-ionic polymers, the removal efficiency for wastewater turbidity, TSS, COD and color were 99.93%, 99.94%, 88% and 94.57%, respectively. The imposed cost for treating one cubic meter of ceramic-tile wastewater treatment by PAC + anionic and PAC + anionic and non-ionic polymers in comparison with PAC alone was reduced to 22.96% and therefore economically more affordable for the tile industry wastewater treatment.Enhanced treatment of ceramic-tile industry wastewater was investigated by modified coagulation–flocculation process using combination of poly Organic materials in wastewater are mainly produced from the additives used in decorating tiles .Characteristics of the ceramic-tile industry raw wastewater are presented in Table 2(OH)nCl6−n. YH2O, Al2O3 = 30%wt, basicity = 65% and pH = 3) was used as a coagulant and anionic (A300), cationic (C270) and nonionic polymers, as the coagulant aids were provided from AquaTech Company, Switzerland.Poly-aluminum chloride . In order to prepare a 0.1 percent solution as coagulant aid, 0.1 g of each polymer was provided separately. Considering the polymers, a 0.1 percent solution was prepared; 0.05 g of each polymer was separately dissolved into 100 ml of distilled water at temperature of 30–50 °C . All experiments were carried out at a temperature of 25 °C. In order to determine the best sedimentation time, 1 L of the wastewater samples was poured into an Erlenmeyer flask at sedimentation time of 10–120 min. Later, the wastewater turbidity and TSS removal were measured to determine the best time of sedimentation before coagulation and flocculation. In order to determine the optimal pH, we used the lime solution and normal hydrochloric acid and adjusted the pH in the range of 5–11. Then, the constant concentration of PAC (200 mg/L) was added to them using the jar test. The optimum pH was determined for each sample by measuring the removal efficiencies of turbidity, TSS, COD, and color  and color were analyzed by using standard methods  , chemical oxygen demand (COD), biochemical oxygen demand (BODods APHA . ParametCH) APHA .Data normality was investigated by Kolmogorov–Smirnov and the homogeneity of data was determined using Levene test. To compare the turbidity, TSS, COD, and color of wastewater with the independent variables such as PH and different concentrations, we run the multivariate analysis (MANOVA) (P = 0.05). The Tukey test was also used to conduct multiple average comparisons between the groups.1 and C0 are the cost and E1 and E0 are the effects in the intervention and control groups, respectively.In order to evaluate the economic efficiency, we used the ICER statistical formula Eq. . In thisThe effects of the primary sedimentation time on the TSS and turbidity removal efficiencies before adding a coagulant are represented in Fig. The results of determining the optimal pH using a fixed dosage of PAC (200 mg/L) are represented in the Fig. The effects of PAC dosage on the removal efficiency of turbidity, TSS, COD and color is shown in the Fig. 300) (0.5 mg/L) was observed 99.66% for turbidity, 99.72% for TSS, 85.12% for COD and 87.2% for colors. The best removal efficiency in combination of PAC (300 mg/L) + cationic polymer C270 was observed in 99.58% for turbidity, 99.64% for TSS, 52.5% for COD and 62.4% for color respectively. The highest removal efficiency in combination of 300 mg/L of PAC with non-ionic polymer were 99.41%, 99.48%, 76.25% and 55.03% for turbidity, TSS, COD and color, respectively. As a result, the applied polymers as coagulant aids decreased the efficient dose of PAC coagulants from 400 mg/L to 300 mg/L.In this stage of experiment, a constant dose of each polymer (0.5 mg/L) was added to different doses of PAC coagulants. The results are presented in Table 300, cationic C270, and non-ionic polymers on removal efficiency of turbidity, TSS, COD, and color are represented in Fig. 300 was 1.5 mg/L with removal efficiency of turbidity, TSS, COD and color 99.87%, 99.89%, 87.5% and 93.02%, respectively. The most removal efficiency was in 2 mg/L cationic polymer for turbidity (99.85%), TSS (99.88%), COD (65%) and color (89.14%). Furthermore, the most removal efficiency of non-ionic polymer for turbidity, TSS, COD and color were 99.68%, 99.73%, 86.5% COD and 84.88%, respectively, in concentration of 2 mg/L. As it is observed in Fig. The purpose of this stage of jar-test experiment was to identify the suitable dose of polymers, which could be used in combination with optimal dose of PAC coagulant (300 mg/L) for treatment of the ceramic wastewater. The applied polymer dose varied from 0.5 to 3 mg/L. The effect of various doses of anionic AAt this stage of the experiment, the anionic, cationic, and non-ionic polymers were combined in different doses of 0.5–3 mg/L. Later, we investigated the effect of this composition combined with the optimal dose of PAC. The effect of various doses of anionic-nonionic polymer combination on removal efficiencies of turbidity, TSS, COD, and color is shown in Fig. 300/nonionic) and PAC + A300 were higher than those of other options. The economic evaluation formula (ICER) was applied to compare the costs of different methods used for ceramic- tile wastewater treatment; the results of which are tabulated in Table The comparisons of removal efficiencies attributed to different coagulant-polymers combinations are indicated in Table Removal efficiency of heavy metals including cadmium (Cd), chromium (Cr), nickel (Ni), zinc (Zn), lead (Pb), and boron (B) in the coagulation–flocculation process under the optimum condition are represented in Fig. In this study, the TSS and turbidity removal efficiencies at the primary sedimentation time of 100 min were 39.9% and 41.97%, respectively. Fahimnia et al.  studied 2+ and Al(OH)2+). So, they neutralize the charges, adsorb the organic pollutants and solids, and consequently increase the removal efficiency , in which charge neutralization and adsorption mechanisms did not happen for the pollutant removal . As a result, application of polymers as coagulant aids decreased the efficient dose of PAC coagulant. Polymers act as aids in cleansing the water and wastewater. They even can be used as primary coagulants for some purposes  showed the highest removal efficiency for turbidity, TSS, COD, and color. Considering the polymers’ combinations, the combination of anionic A300 and non-ionic polymers in the optimal dose had the highest removal efficiency. The treatment cost for one cubic meter of ceramic-tile wastewater using the PAC + anionic as well as PAC + anionic and non-ionic polymers was 22.96 percent less than the PAC-alone method.Finally, the results of this study show that the coagulation–flocculation process as well as the PAC combination with anionic, cationic, and nonionic polymers can be used as an effective method for treatment of ceramic-tile wastewater. The PAC, as the coagulator with the optimal dose of 300 mg/L and the anionic polymer A


Adsorption performances of Cu(II), Cr(VI), and Fe(III) on CAs well fitted with a pseudo-second-order kinetic model. The structures and morphologies of metal-containing samples were characterized by scanning electron micrographs (SEM), Energy Dispersive Spectrometer (EDS), transmission electron microscope (TEM), and X-ray diffraction (XRD). The results demonstrated that the texture and electrochemical performance of CAs adsorbing metal ions exhibited a clear change. The specific surface area of CAs for adsorbing copper ions was 450 m2·g−1 and they showed a small average pore diameter (7.16 nm). Furthermore, CAs adsorbing metals could be used for the super capacitor. The specific capacitance of CAs adsorbing copper ions could reach 255 F·g−1 at a current density of 1.0 A·g−1. The CA-Cu electrode materials exhibited excellent reversibility with a cycling efficiency of 97% after 5000 cycles.Environmental problems caused by metal ions have caused widespread concern in recent years. In this work, carbon aerogels (CAs) adsorbing different metal ions were prepared. The adsorption performance and kinetics of metal ions (Cu(II), Cr(VI), and Fe(III)) on carbon aerogels were systematically investigated. The results indicated that the maximum adsorption capacity of Cu(II) was 424 mg·g Water pollution and soil contamination from heavy metal ions, which resulted from various industrial manufacturing, had been a worldwide environmental problem. Therefore, the removal of heavy metal ions from wastewater and soil have attracted more and more attention in recent years. Traditional methods for removing heavy metal ions included adsorption, precipitation, flocculation, reverse osmosis, ion exchange, and electrochemical treatments ,2,3,4,5.−1 and the removal rate was 96.03%. Li et al. [−1 for copper ions by CAs in aqueous solutions in the presence of a surfactant. Wang et al. [Pekala et al.  first dii et al.  found thg et al.  found th3 and N2. The results showed that the specific capacitance of the CF@VN material was 104.05 F·g−1 at the current density of 0.5 A·g−1. Hao et al. [−1 at 2 mV·s−1, respectively. Yu et al. [The super capacitors have been widely used and studied because of their advantages such as long service life and high power density ,14. CAs o et al.  used chiu et al.  reduced −1 at 1.0 A·g−1. Carbon aerogels as adsorbent provided a new way to remove metal ions. The corresponding materials exhibited an excellent electrochemical performance and could be used as super capacitor electrode materials and resource utilization.In this case, carbon aerogels were used to adsorb Cu(II), Cr(VI), and Fe(III). The adsorption isotherms were suitable for the Langmuir model. In addition, metal-adsorbed carbon aerogels could be used as super capacitor materials. By observing electrochemical measurements, the specific capacitance of carbon aerogels adsorbing copper ions could reach 255 F·gPhloroglucinol (P), resorcinol (R), and formaldehyde (F) were used as raw materials to prepare the carbon aerogels. The molar ratio of P/R was 0.14. In addition, sodium carbonate (C) and ascorbic acid (Vc) were used as catalysts. The ratios of C/(P + R) and Vc/(P + R) were 0.002 and 0.02, respectively. The mass concentration of the distilled water (P + R + F + C + Vc) in the solution was 45%. All of the solutions of raw materials were prepared in distilled water and stored in glass bottles. Afterward, the glass bottles were placed into an ultrasonic bath to obtain uniform solutions. The bottles were left in the water bath at 60 °C for 2 days, which generated gels. The wet gels were placed in acetone for 3 days and the solvent was replaced every day. The gel was placed in a tube furnace, evacuated, and then purged with nitrogen. Afterward, the gels were carbonized by a temperature programming method. The temperature of the carbonation process was set so that uniform holes would form. Lastly, the CAs were obtained.4)2Fe(SO4)·6H2O was dissolved in distilled water to prepare a standard iron solution. The standard iron solution  and 5 mg of CAs were added to a beaker and stirred on a magnetic stirrer for 1 h, 2 h, 3 h, 5 h, 7 h, 9 h, 11 h, and 18 h. The stirred solution was centrifuged, 20 mL of the supernatant was transferred to a 100 mL volumetric flask, and then it was diluted with water to the mark. The absorbance was measured at its maximum absorption wavelength (230 nm) and the concentration of Fe(II) versus absorbance was plotted [2CrO4) was almost the same as that of Fe(III). However, Cr (VI) was measured at a maximum absorption wavelength of 370 nm [3)2·3H2O), the copper reagent, which served as the color developing agent was added, and the pH was adjusted to 9 with ammonia (13 mol/L). The concentration was determined at the maximum absorption wavelength (452 nm) [Fe(II) was determined by spectrophotometry. A total of 4.21 g  and the mixtures were stirred for 24 h so that the metal ions were fully adsorbed and then the samples were dried in an oven at 45 °C. Lastly, the samples were carbonized at 600 °C. The CAs alone were used as the contrast. We named the carbonized samples as CA-Fe, CA-Cr, CA-Cu, and blank, respectively, and they were denoted CA-X and the blank. Additionally, the CAs adsorbing copper ions that were not carbonized were denoted CA-Cu-0. After carbonization, all the samples were ball milled for 6 h and then passed through a 200 mesh sieve. The samples were then added to polytetrafluoroethylene (PTFE) at a mass ratio of 95:5. Approximately 40 mL of deionized water was added and the mixtures were stirred for 6 h and dried in a drying oven at 105 °C overnight. Lastly, the dried substances were placed on foam nickel. The electrodes were prepared by using a tablet press with 10 MPa. Then the samples were dried and weighed. To test the electrochemical performance, the samples were soaked in 6 M KOH electrolyte for 24 h.The CA powder (3 g) was mixed with 200 mL of iron solution, chromium solution, and copper solution  and k2 (g·(mg·min)−1) stood for the rate constants of pseudo-first-order and pseudo-second-order adsorption, respectively. t was the reaction time (min), t (mg·g−1), and eQ was the balanced adsorption capacity (mg·g−1). In Equation (1), plotting In the equation, In 1918, Langmuir  proposedeQ is the balanced adsorption capacity (mg·g−1), mQ is the monolayer saturated adsorption (mg·g−1), b is the adsorption equilibrium constant (L·mg−1), and Ce is the concentration of adsorbates in the system at the adsorption equilibrium (mg·L−1).In this equation, eQ). The amount of adsorbed metal was calculated by the following formula [In addition, the adsorption capacity of carbon aerogels could be assessed based on the amount of adsorbed metal ( formula .eq is the adsorption capacity (mg·g−1), C0 and eC are the concentrations of metal ions in the solution before and after adsorption, respectively (mg·L−1), m is the quantity of adding adsorbent (g), and V is the volume of the solution containing metal ions (L).In Formula (4), 2. The surface morphologies of the samples were analyzed by SEM , which were equipped with an APOLLO EDS immobilizing the sample with conduction glue. The internal morphology was observed by TEM , which the ground CA-X powder was placed in an absolute ethanol solution and the ultrasonic wave was evenly uniform and dropped into the micro-grid. XRD  was used to identify the specific peak to metals and the peaks characteristic of the CAs. The surface compositions of CA-X were characterized by XPS . A UV-vis spectrophotometer  was used to measure the metal ions concentration and characterize the adsorption capacity of CAs. Electrochemical performances were measured with a GAMRY Interface 1000 electrochemical workstation. All the measurements used a three-electrode system with a saturated calomel electrode (SCE) and Pt as a reference and counter electrode. The CA-X and blank samples were the working electrode. In this work, cycle voltammetry , electrochemical impedance spectroscopy (EIS), and charge and discharge testing  were used for the investigation. The formula to calculate the specific capacitance was shown below [The gels were carbonized by tube furnace  at 600 °C. The Brunaure-Emmett-Teller method and Density-Functional-Theory (DFT) model calculated the specific surface area and the micropore surface area . The pore volumes and pore size were used in the Barrett-Joyner-Halenda (BJH) model. The degassing temperature was maintained at 90 °C for 1 h and then increased to 250 °C for 6 h. The carrier gas was Nwn below .mC was the single-electrode specific capacitance (F·g−1), I was the charge current (A), ∆t was the charge time (s), m was the quality (g), and ∆V was the potential window (V).In this case, 2) of CA-Fe, CA-Cr, and CA-Cu were 0.997, 0.946, and 0.915, respectively. In the pseudo-second-order kinetic model . The adsorbing metal ions followed pseudo-second-order kinetics model.The results of the pseudo-first-order kinetic and the pseudo-second-order kinetic models under different stirring time were shown in ic model B, the R22 = 0.9999), which indicated that the adsorption process of Cu(II) on CAs was well fitted with physical adsorption. The adsorption capacities of carbon materials were largely dependent on the surface functional groups such as –COOH and –C=O, which could be introduced on the surface of the CAs. These functional groups mainly contributed to the adsorption of Cu (II) on the surface of the CAs [The results of fitting the Langmuir isothermal adsorption equation to the experimental data were shown in  the CAs . Further the CAs . Therefo−1. As shown in −1 and 139 mg·g−1, respectively. The reason for this was the strong electrostatic attraction between the metal ions and the CAs [3 was added to dissolve the (NH4)2Fe(SO4)·6H2O and the pH was approximately 6. The pH value of the standard chromium solution was 7 by using HNO3 and NaOH to adjust. In acidic solutions, metal ions competed with H+ for the adsorption sites, which resulted in the reduction of adsorption of the heavy metal ions. Therefore, Fe(III) had the smallest adsorption capacity (133 mg·g−1) and the adsorption amount of Cr(VI) was 139 mg·g−1. However, in an alkaline solution, the proportion of (CuOH)+ was large, the average charge density was small, and the electronegativity was high. At the same time, more anion adsorption sites provided by functional groups such as carboxyl and carbonyl and surface complexation with Cu(II) resulted in increasing adsorption [ the CAs . Before sorption . The max2 adsorption-desorption isotherms of CA-Fe, CA-Cr, CA-Cu, and the blank at 77 K were shown in 0 > 0.8, all the curves had an H1 type hysteresis loop, which indicated that there were mesopores and macropores in the materials and capillary condensation occurred.The NBET), total pore volume , micropore surface area (Smicro), and average pore diameter (Daverage) of the CA-Fe, CA-Cr, CA-Cu, and blank samples. Clearly, the CA-Cu sample had the smallest SBET, Vtotal, Smicro, and Daverage values. The result was attributed to the occupation of adsorbing metal ions on CAs. As presented in 2 adsorption-desorption isotherms discussion. At the same time, the decreasing tendency of the specific surface area showed that the metal ions adsorption process was physical adsorption.θ = 30.1°, 35.8°, 43.1°, and 53.8° could be indexed to the (220), (311), (400), and (422) planes of Fe3O4, respectively [2O3 [ely [2O3 ,39. Rega1s and O1s from 1s spectra possessed three peaks at 531 eV (C=O), 533.3 eV (C-O-C), and 532 eV (COOH). The C1s spectrum could be fitted into six peaks at 284.6 eV, 285.3 eV, 287.4 eV, 285.4 eV, 289.05 eV, and 291.7 eV corresponding to SP2, SP3, C=O, C-O, O-C=O and π-π, respectively [ectively ,41. It wectively .−1 were shown in The electrochemical properties of CA-Fe, CA-Cr, CA-Cu, and the blank were investigated in a three-electrode system by using 6 M KOH aqueous as the electrolyte. The CV curves acquired at a scan rate of 0.5 mV·s−1. The curves were similar to isosceles triangles, which indicated that the charge-discharge had good symmetry and linearly increased or decreased over time. This shape also suggested that this material could serve as a reversible double layer capacitor. In addition, the ion hydration radius (γH) was related to the diffusion resistance. γH of Cu(II) was 4.19 Å, γH of Cr(VI) was 4.61 Å, and the γH of Fe(III) was 4.67 Å [γH of Cu(II) was the smallest. Smaller γH values indicated lower diffusion resistance, which corresponded to improve charge transfer. The specific capacitance of the CA-Cu was 255 F·g−1, the blank was 125 F·g−1, and the CA-Fe was 215 F·g−1. The specific capacitance data were shown in −1, which was higher than that of CA-Cu-0. This result showed the copper ions of the CA-Cu-0 could desorb and come into the electrolyte. s 4.67 Å ,44. Clea−1. It revealed that the specific capacitance of CA-Cu still remained at 227 F·g−1 (~97%) after 5000 charge-discharge cycles. Clearly, the CA-Cu displayed good cycling stability. Compared with CA-Cu-0, the CA-Cu had a better stability. At the same time, when compared with CA-Fe and CA-Cr, the better stability features of the CA-Cu might be the compact structures.ct). The low frequency region reflected the ion diffusion rate, Rb was the equivalent circuit resistance, Cdl was the stationary phase element, Zw was the Warburg impedance, and CL was the intercalation capacitance. The electrode kinetics characterized the effect of metal adsorption on the pore structure. The EIS method evaluated the apparent chemical diffusion (Dk+) of the blank and adsorbed metal into the CAs. The K+ diffusion coefficient was expressed by the following formula [ formula .σ is the slope of the line and ω is the angular frequency in the low frequency region. In Equation (7), f is the frequency. In addition, A is the specific area of the electrode (0.785 × 10−4 m2), F is the Faraday constant (9.65 × 104 C·mol−1), n represents the number of electrons per molecule involved in the electron transfer, C is the concentration of K+ in the KOH solution (6 × 103 mol·m3), T is the experimental temperature (298 K), and R is the gas constant (8.314 J·K−1·mol−1). + diffusion coefficients in the CA-Fe, CA-Cr, CA-Cu and blank samples are 0.76 × 10−15, 1.8 × 10−15, 8.0 × 10−15 and 0.5 × 10−16cm2·s−1, respectively. It could be found that the Dk+ values of CA-X and blank are in the magnitude of 10−15 cm2·s−1 and the diffusion coefficients of K+ are increased when the Rct increases. This phenomenon revealed that the kinetics of K+ and electro transfer into the electrodes were much faster at low Rct. Among these samples, K+ diffusion coefficients in the CA-Cu was the biggest. Therefore, the charge transfer resistance of CA-Cu was the smallest.In Formula (6), −1. The dynamics equation confirmed to the pseudo-second-order model and adsorption behavior could be described by the Langmuir adsorption isotherm equation, which indicated the adsorption process was single layer adsorption. The structure of CA-X could be more compact. In addition, metal-adsorbed carbon aerogel materials could be used as super capacitors. According to the results of electrochemical measurement, the electrochemical performance of CAs with adsorbed Cu(II) exhibited the highest specific capacitance, which reached 255 F·g−1 at 1.0 A·g−1. The excellent adsorption performance indicated that the carbon aerogel was a potential and effective absorbent for metal ions removal and the corresponding carbon aerogels could be used for super capacitors.In summary, CAs with a higher specific surface area were synthesized through the sol-gel process and the prepared materials exhibited an excellent adsorption capacity. At the same time, the metal ions adsorption on carbon aerogels executed a noticeable improvement for high super capacitor performance. In the adsorption tests, CAs demonstrated a high adsorption capacity for Cu (II), which could reach 424 mg·g


An important question in neural correlate of consciousness (NCC) studies is whether event-related potential (ERP) component P3 reflects visual awareness or the confidence with which one reports a visual experience. In the present study, participants detected visual stimuli presented at threshold-level contrast, then rated their subjective confidence with respect to their response on a four-point scale . Because awareness responses in trials with rating of “not confident at all” were likely noise, we analyzed the data excluding those trials. The ERP results revealed a significant positive difference in P3 amplitude between “aware” and “unaware” trials. P3 amplitude was more positive in aware trials compared to unaware trials. Importantly, this pattern was observed for trials with combined confidence ratings of “very confident” and “quite confident,” and for trials with confidence ratings of “slightly confident,” suggesting that awareness alone can modulate P3. A significant interaction between awareness and confidence is reported, suggesting that confidence influences P3 as well. In addition, ERP results revealed that visual awareness negativity (VAN) was observed over posterior temporal and occipital electrodes and largely not influenced by confidence. This result indicated that VAN is an early neural correlate of visual awareness. The neural correlate of consciousness (NCC) is defined as “minimum neural mechanism jointly sufficient for any one specific conscious experience” , p. 307.Over the past 20 years, most ERP studies on NCC focused on visual processing. In particular, researchers recorded ERP during the presentation of a threshold-level visual stimulus. The presentation of such weak stimuli ensured that on some trials participants were aware of the stimulus, and on other trials they were not. Researchers then contrasted brain ERP’s from “aware” trials with “unaware” trials for the same stimulus intensity  , allowinBy using such contrastive experimental designs, two candidate electrophysiological NCCs have emerged: visual awareness negativity (VAN) and late positivity (LP). VAN is the negative amplitude difference between aware and unaware conditions . It typiHowever, it has been argued that the P3 may reflect variations in participants’ confidence with respect to their response . BecauseDespite the findings reported by Given the debate in the literature , the aimWe used a forced-choice detection task, modified after that of M = 22.25, SD = 2.59). All participants had normal or corrected to normal eyesight; none of them had history of neurological disease or brain injuries. All participants gave written informed consent prior to the study. All procedures were approved by the Ethics Committee in Academy of Psychology and Behavior, Tianjin Normal University.Thirty-one right-handed undergraduate students participated in this study. However, 15 participants were excluded. Among them, 4 participants reported awareness in less than 25% of the critical trials or more than 75% of the critical trials and 11 participants did not have enough (>20) critical trials for each awareness/confidence combination used in the data analysis . The final sample consisted of 16 participants , tilted 45 degree to the left.Stimuli were controlled with E-prime software on a 1024 × 768 resolution monitor with 60 Hz refresh rate. The luminance of the gray background was 22 cd/m”  at the center of the screen for 1200 ms. Next, a black fixation cross was presented for 1200 ms, followed by the stimulus . The fixation cross and stimulus were presented in the center of the screen. Stimulus duration was calibrated to each participant’s perception threshold (see below). After the stimulus (or blank), the participants indicated whether they had seen the stimulus by pressing one of two designated buttons using the same hand. Finally, a prompt appeared on the screen asking participants to rate their subjective confidence with respect to their response on a four-point scale  was used than in the critical trials.In pre-experimental calibration phase, the stimulus presentation was identical to that in the main experiment. The participants were instructed to indicate whether they had seen the stimulus in each trial, but they were not instructed to rate their subjective confidence. In the first calibration block, the duration of the critical stimuli was two frames. Each calibration block included 20 critical trials, 5 catch trials, and 5 control trials. In next block, the number of screen refreshes was increased or decreased by one frame according to the participants’ performance. If the participants reported awareness in less than 25% or more than 75% of the critical trials, the number of refresh frames was increased or decreased with one frame correspondingly. When the appropriate stimulus duration was found, the same calibration block was repeated once again to make sure that performance remained stable.The EEG was recorded from 64-channel Ag/AgCl electrode cap  according to the 10–20 system. Vertical EOG recording electrodes were positioned above and below the left eye, and horizontal EOG recording electrodes were positioned 1.5 cm from the outer canthus of each eye. The reference electrode was placed on the nose. The ground electrode was placed in front of Fz. The EEG and EOG signals were amplified with a band pass of 0.05–400 Hz and sampled at 1000 Hz. The impedance was kept below 5 kΩ.Electro-encephalography data were analyzed offline with Curry 7. Offline correction of eye movement artifact was performed. To exclude trials contaminated by artifacts, trials with voltages exceeding ± 80 μV at any electrodes were discarded. The signals were averaged offline over 800 ms periods, and an additional 100 ms was recorded prior to the probe onset to allow for baseline correction. The data were filtered with 0.1 Hz high pass and 30 Hz low pass filters.M = 71, SD = 37.1), aware/slight-confidence , unaware/high-confidence , and unaware/slight-confidence .In terms of confidence ratings, participants rarely chose “very confident.” For this reason the data from “very confident” trials and “quite confident” trials were combined into a high confidence dataset to increase statistical power. Additionally, the data from “not confident at all” were eliminated from ERP analyzed. Therefore ERPs for four types of critical trials were obtained : aware/high-confidence (Based on previous similar studies  and inspn = 16) reported awareness in 41.02% (SD = 13.37) of the critical trials; this level of awareness was appropriate, as our goal was to obtain about 50% awareness. The proportions of response for each awareness/confidence combination are presented in Participants (n = 16) reported awareness in 97.66% (SD = 3.77) of the control trials, and unawareness in 88.49% (SD = 17.32) of the catch the trials. Overall the participants indeed followed the instructions.In addition, participants , posterior temporal , and occipital  electrodes are shown in F = 4.18, p > 0.05, F = 13.11, p < 0.05, Analysis of variance on average N1 amplitudes showed that main effect of awareness was not significant  and occipital electrodes (-0.54 ± 0.97 μV vs. 0.52 ± 0.65 μV) , but not over the parietal electrodes (0.97 ± 0.81 μV vs. 1.57 ± 0.6 μV) . Other interactions involving Awareness were not statistically significant (ps > 0.05).The interaction between awareness and confidence was not significant [.21] see . In addiF = 3.93, p > 0.05, F = 3.03, p > 0.05, Analysis of variance on average N2 amplitudes showed no significant main effect of awareness  and occipital electrodes (0.96 ± 1.08 μV vs. 2.57 ± 0.79 μV) , but not over the parietal electrodes (3.46 ± 1.02 μV vs. 4.23 ± 0.78 μV) . Other interactions involving Awareness were not statistically significant (ps > 0.05).The interaction between awareness and confidence was not significant [.03] see . There wF = 19.65, p < 0.01, F = 1.19, p > 0.05, F = 11.04, p < 0.01, Analysis of variance on average P3 amplitudes showed a significant main effect of awareness . Notably, when the participants indicated slight confidence, there was also a significant difference in P3 amplitude between aware trials and unaware trials (8.55 ± 1.02 μV vs. 6.93 ± 0.97 μV)   [.05] see .F = 5.58, p < 0.05], but, for unaware trials, the different in P3 amplitude between high confident trials and slight confident trials did not reach statistical significance (6.44 ± 0.93 μV vs. 6.93 ± 0.97 μV) .Simple-effect analysis also showed that, for aware trials, P3 was more positive in high confident trials compared to slight confident trials (10.09 ± 1.35 μV vs. 8.55 ± 1.02 μV) , posterior temporal electrodes (8.67 ± 1.01 μV vs. 6.58 ± 0.79 μV) , and occipital electrodes (8.3 ± 1.25 μV vs. 5.77 ± 1.01 μV) . Other interactions involving Awareness were not statistically significant (ps > 0.05).There was also a significant interaction between awareness and areas .Finally, we have run an additional ANOVA using our entire dataset including data from The aim of the current study was to investigate whether P3 reflects visual awareness and/or confidence. For the most part of the discussion, unless we specified, we will discuss results from the data excluding “not confident at all” trials. We found P3 amplitude to be more positive in aware trials compared to unaware trials (evidence for LP). Moreover, we observed a difference in P3 amplitude between aware trials and unaware trials  for trials responded with slight confidence as well as high confidence.The main finding of the present study is a positive difference in P3 amplitude between “aware” and “unaware” trials for both confidence levels, indicating a robust effect of awareness on P3 amplitude. Critically, our results differ from that obtained by all four confidence levels implemented in our study . Indeed when the “not confident at all” trials were included there was no significant difference in P3 amplitude between “aware” and “unaware” trials in low confidence trials.In fact, the reason The second piece of our results is that for P3 amplitude, we found significant interaction between awareness (aware vs. unware) and confidence (high vs. slight confidence) even after we removed the data for “not-at-all confident” trials. This suggests that even though P3 is clearly modulated by awareness, confidence might also contribute to the P3 waveform. However, the effect of confidence found here should be interpreted with caution. It is conceivable that given the low level of confidence rating in the “slightly confident” trials, there could be some noise in the awareness response and consequently led to the effect of interaction .In the literature, researchers often argue whether the P3 reflects awareness  or confiOur results point to the conclusion that awareness modulates P3. But the nature of difference between aware vs. unaware responses remains unanswered. One might argue that in contrast to aware responses, unaware responses could be caused by neural noise which exceeds the signal strength. Fluctuations in attention level at the moment of encoding could also result in sensory signals being amplified on some trials  but not others .There have been some concerns that NCC studies using contrastive experimental designs not only reveal processes directly corresponding to conscious experience, but also other processes that precede conscious perception  or follow conscious perception (i.e., post-perceptual processes; Another main finding of the present study is that VAN is likely an early neural correlates of visual awareness. Our results showed a significant N1 difference between aware and unaware trials over posterior temporal and occipital electrodes. Similarly, there was a significant N2 difference between aware and unaware trials over posterior temporal and occipital electrodes. According previous studies e.g., , this siOur finding is in line with previous studies e.g., , which sAs shown in Overall, our results revealed a significant positive difference in P3 amplitude between “aware” and “unaware” trials that was found both in high confidence trials and slight confidence trials. This empirical trend supports the idea that P3 reflects awareness. Moreover, we found a significant interaction between awareness and confidence ratings, suggesting that confidence could modulate P3 as well. Finally, we detected VAN over posterior temporal and occipital electrodes suggesting that VAN over those electrodes is an early neural correlates of visual awareness.This experiment was approved by the Ethics Committee in Academy of Psychology and Behavior, Tianjin Normal University. All participants gave written informed consent in accordance with the 2013 Declaration of Helsinki and were paid for their attendance.MY, YL, and H-JS designed the experiments. MY and YL prepared the materials and performed the experiments. MY, YL, BS, and H-JS analyzed the data and wrote the manuscript. All authors approved the final version of the manuscript for submission.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The results showed that TS could significantly enhance the absorption of SA5, SA6, and DISS monomers. Verapamil, a P-glycoprotein inhibitor, was able to elevate the absorption of SA5 and SA6, and an absorption experiment using Rho123 led us to conclude that TS influenced the absorption of SA5 and SA6 in a manner similar to that of a P-glycoprotein inhibitor. Sodium caprate, a paracellular absorption enhancer, was found to increase the absorption of SA5, SA6, and DISS. Results showed that the absorption mechanisms of SA5 and SA6 may combine active transport with paracellular passive penetration, while DISS's absorption was dominated by paracellular passive penetration. However, the relationship between polygala saponins and the absorption of SA5, SA6, and DISS by paracellular passive penetration remain to be examined. This is the direction of our future research.Oligosaccharide esters, which are among the main active components of Polygalae Radix (PR), demonstrate significant pharmacological activities in the human nervous system. In our previous research, some other constituents in PR were able to improve the bioavailability of oligosaccharide esters such as sibiricose A5 (SA5), sibiricose A6 (SA6), and 3,6′-disinapoyl sucrose (DISS), but the related components and their underlying mechanisms remain unknown. The present study aimed to investigate the intestinal absorptive profile of SA5, SA6, and DISS and the absorptive behavior influenced by the coadministration of polygalaxanthone III and total saponins of PR (TS) using an Polygala tenuifolia Willd. or Polygala sibirica L. It has been used for many years for the treatment of amnesia, neurasthenia, palpitation, insomnia, and depression [β-amyloid (Aβ) [Polygalae Radix , one of the commonly used traditional Chinese medicines (TCM), is derived from the roots ofpression . Pharmacoid (Aβ) , antioxioid (Aβ) , antidepoid (Aβ) –6, and noid (Aβ) , 8. The oid (Aβ) –11. Amonoid (Aβ) –15. Sibioid (Aβ) . These toid (Aβ) –19. Otheoid (Aβ) , 21. In oid (Aβ) , 23. How in vitro. Then, the influence of polygalaxanthone III (PT) and total saponins (TS) on the absorptive profile of SA5, SA6, and DISS and their possible influence mechanism were investigated. Samples were collected from the sac contents and were analyzed for three oligosaccharide esters using RP-HPLC.The efficacy of TCM is based on the active ingredients, and the various effective components are combined in proportion to exert efficacy. With the further development of modern studies on TCM, development of modern Chinese herbal drugs by studying compatibility of various effective components in single Chinese medicines or compounds has become a key research direction. Studies on the synergy between multiple ingredients in one type of TCM could also help to reveal the mechanisms of the integral effects of any particular TCM. Therefore, in order to explore the synergistic mechanism between multiple components in RP from the point of intestinal absorption, the present research was designed to study the intestinal absorptive profile of three oligosaccharide esters  Base in Heyang City, Shaanxi Province. The roots were identified by Yuting Chen (Beijing University of Chinese Medicine). Sibiricose A5 and sibiricose A6 were isolated from Polygala tenuifolia in our laboratory [1H-NMR, and 13C-NMR) and their purity was determined by HPLC (purity ≥ 96%). Polygalaxanthone III (purity ≥ 98%) was purchased from Nanchang Beta Biological Technology Co., Ltd. . We purchased 3,6′-disinapoyl sucrose and verapamil from the National Institutes for Food and Drug Control, Beijing, China (purity ≥ 96%). Rhodamine 123 (Rho123) and sodium caprate were obtained from Sigma Chemical Co. . The culture solution was a Krebs–Ringer (K–R) culture solution, containing 3.90 g NaCl, 0.175 g KCl, 0.685 g NaHCO3, 0.01 g MgCl2, 0.16 g NaH2PO4, and 0.35 g glucose in 500 mL distilled water. MCI GEL CHP20P resin was purchased from Mitsubishi Chemical Corporation (Japan).The roots ofboratory . Their sHPLC-grade acetonitrile and methanol were obtained from Fisher Scientific Inc. . Purified water was purchased from Hangzhou Wahaha Group Co., Ltd. . Oxygen-generating agent was purchased from Oxygen Stand Firm . All of the other chemicals were of analytical grade.SD rats  were obtained from the Vital River Experimental Animal Co., Ltd. . The experiments on animals were in conformity with the Guidelines of “Principles of Laboratory Animal Care”  and were approved by the Animal Experimentation Ethics Committee of Capital Medical University. The rats were housed in a temperature- and humidity-controlled room  with standard diet and water. Prior to the experiments, the rats were acclimatized for three days and had free access to water but no food for 12 h before the experiments.2O, 40% EtOH, and 80% EtOH to afford three fractions. The 80% EtOH fraction was concentrated under reduced pressure to obtain the extract of TS from Polygalae Radix (9.40 g).PR (400 g) was refluxed with water  for 2 h twice. Afterwards, removal of the solvent under reduced pressure was performed, resulting in an extract of Polygalae Radix (PRE) weighing 143.12 g. The PRE, which was analyzed by LC, contained 0.417% SA5, 0.716% SA6, 1.65% DISS, and 0.099% PT. PRE (71.56 g) was further applied to column chromatography on MCI GEL CHP20P resin eluting with Hμm, 250 mm × 4.6 mm) protected with an Xbridge™ RP18 guard column . The mobile phase consisted of CH3CN (A) and water containing 0.1% (v/v) phosphoric acid (B). A gradient program was used as follows: 8%–14% A (0-15 min), 14%–18% A (15–25 min), 18%–21% A (25–35 min), 21%–25% A (35–40 min), 25%–27% A (40–46 min), 27%–30% A (46–53 min), 30%–35% A (53–62 min), 35%–40% A (62–75 min), and 40%–55% A (75–85 min). The flow rate was 1.0 ml/min. The column temperature was 30°C. The detection was performed at a wavelength of 330 nm. PRE was dissolved in K-R culture solution at concentration of 2.07 mg/ml. Working standard solutions of SA5, SA6, and DISS in the concentration ranges of 1.09–130.20 μg/ml, 2.25–270.00 μg/ml, and 2.66–212.80μg/ml, respectively, were freshly prepared in K-R culture solution. A calibration plot was obtained by plotting SA5, SA6, and DISS peak areas against concentration; a regression equation was obtained by linear regression analysis. The intraday precision was determined within one day by analyzing six replicates of samples of sac contents at concentrations of 50 μg/ml (SA5), 86 μg/ml (SA6), and 198 μg/ml (DISS). The interday precision was determined on the third day of analysis for the samples of sac contents. The intraday and interday precision was defined as the relative standard deviation (RSD). The stability of PRE samples at room temperature for 24 h was determined six times for the sac contents. The average recoveries were determined by analyzing the PR sample solutions mixed with corresponding referenced standard six times, and we took the average expressed as a percentage value .Regarding the detection of SA5, SA6, and DISS, the HPLC system consists of a Waters 2695 HPLC pump, a Waters ASI-120 automated sample injector, and a Waters 2996 Photodiode array detector . Samples were separated on a Waters Xbridge™ shield RP18 column  was used for the detection of Rho123 analysis at an excitation wavelength of 500 nm and an emission wavelength of 525 nm. The regression equation was Y = 298.34X - 2.0279; r = 0.9991.For determination of Rho123, it was prepared in K–R culture solution at the concentrations of 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, and 1.5 μm, 150 mm × 4.6 mm). The mobile phase consisted of CH3CN (A) and water (B). A gradient program was used as follows: 5%–12% A (0–5min), 12%–25% A (5–10 min), 25% A (10–18 min), 5%–30% A (18–23 min), 30%–45% A (23–33 min), and 45%–70%A (33–40 min). The flow rate was 0.4 ml/min. The column temperature was 25°C. The detection was performed at a wavelength of 330 nm. The mass spectrometer was operated in negative ion mode. The capillary voltage was 5 kV. The dryer flow rate was 6 L/min and the dryer pressure was 2.0 bar at 180°C. The balance plate effect was -500 V.Regarding identification of chemical components in TS, the analysis was performed on an Agilent 1260 series  coupled with a Bruker FT-ICR-MS solariX MALDI/ESI 9.4T . Samples were separated on an Agilent Poroshell 120 SB-C18 column , SA5 , SA6 , DISS , PT (0.012 mg/ml), and TS (1.58 mg/ml) were dissolved in the intestinal culture solution as test solutions. The medium concentration of PRE was equal to the human clinical dose. The concentrations of SA5, SA6, DISS, and PT were calculated by equal amount of PRE. The concentration of TS was calculated according to the amount of raw drugs of PRE.During the2 and 95% O2. Aliquots of 1 ml of serosal solution were taken out from the intestinal sac at different time intervals , and 1 ml drug-free culture solution was injected into the intestinal sac to complement the solution. After sampling, the length and width of the intestinal segments were measured. Then uptake per unit area was calculated.The rats were anesthetized with an intraperitoneal injection of 2% phenobarbital sodium (0.4 mg/kg). After the abdominal area had been shaved, we opened the abdominal cavity with a midline incision (2–3 cm). The jejunum segment of the intestinal sac was isolated carefully and then flushed with ice-cold K-R solution immediately. We removed the underlying mesentery. The intestinal segments were cut into 10 cm lengths, everted carefully with a smooth glass rod, and tied at the outlet at one end. Then, 2 ml of drug-free K-R solution was injected into the serosa compartment and the other end was tied. The everted intestinal sac was suspended in the experimental test solution (25 ml) and maintained at 37°C. The experimental solution was continually aerated with 5% COμm Millipore membrane for HPLC analysis.The samples were mixed with acetonitrile at a ratio of 1:0.2, vortex-mixed for 2 min, and centrifuged at 9,000 rpm for 5 min. The supernatant was obtained and filtered through a 0.45  Q = 2CnVn + ∑i=1n−1CiVi. The sampling time was used for linear regression of the absorbed dose per unit area, and the absorption rate constant Ka was obtained from the slope. The apparent permeability coefficient (Papp) is calculated as Papp = △Q/△t/A/C0, where ΔQ is the amount of drug permeated in Δt (μg), A is the area of the intestinal membrane, and C0 is the initial concentration. Percentage of absorption (P%) = drug concentration in the intestinal sac/drug concentration in the bath × 100%.The cumulative absorption of drugs is described by the equation SD and are analyzed by one-way analysis of variance (ANOVA) and LSD test using SPSS ver. 16.0 software. P values lower than 0.05 were accepted as statistically significant.All of the values are expressed as the mean ± Y = 15293X – 608.48 , Y = 7089.4X - 3321.1 , and Y = 18814X – 37768 . Both the stability and the precision of the analytical method were below 2% (n = 6). Their mean recoveries were 97.45%, 97.67%, and 97.94%. These good results showed that the method was both stable and suitable for continuous monitoring of SA5, SA6, and DISS. According to the literature [Representative chromatograms obtained from PRE solution, the sac absorption solution of PRE, and the reference solutions of SA5, SA6, and DISS are shown in terature , the finThe stabilities of SA5, SA6, and DISS in the blank intestinal circulation solution were RSD = 1.70%, RSD = 0.87%, and RSD = 1.96%, respectively, which showed that they were stable at 37°C for 3 h. This study was designed to ensure that the loss of the drug during the absorption experiment was due only to absorption and not other means.In the present study, the total saponins from Polygalae Radix (TS), which mainly contains polygala saponins analyzed by HPLC-FT-ICR/MS, were prepared. r > 0.9) with time in three concentrations in the everted rat gut sac model. However, there was no statistical difference in Ka of SA5 and SA6 between medium and high concentrations, and the intestinal absorptive profiles of the two concentrations were similar. Moreover, Papp and P% of SA5 and SA6 in the small intestine presented a trend of increasing first and then decreasing, which is different from the characteristic of that in PRE (Ka of DISS increased linearly (r > 0.9) with concentration, and Papp and P% of DISS were unaffected by concentration. The phenomenon suggested that the absorption mechanism of DISS monomer may involve passive diffusion.As shown in t in PRE . These rr > 0.9) with both concentration and time in low, medium, or high concentrations of PRE in the everted rat gut sac model.  Ka, Papp, and P% of SA5, SA6, and DISS monomers are all significantly (P < 0.01) less than that of PRE-treated gut sacs in the three concentrations. These phenomena were consistent with the results of the pharmacokinetic study. Notably, and different from what we observed with SA5 and SA6 monomers, Ka's of SA5 and SA6 in PRE at high concentrations were higher (P < 0.01) than those at medium concentrations. For SA5 and SA6, Papp was significantly increased in both medium and high concentrations compared to their Papp in low concentrations, but there was no significant difference between Papp at medium and high concentrations. Papp of DISS in PRE remains basically unchanged with increasing drug concentration. These results suggested that the absorption enhancement of SA5, SA6, and DISS in PRE and the difference of absorptive profile between the three monomers in PRE versus these monomers alone may be related to the changes in intestinal microenvironment in rat gut induced by other chemicals in the PRE. However, the related ingredients and the underlying mechanism have so far been unclear.In this study, three different concentrations of PRE were used to investigate the intestinal absorption profile of SA5, SA6, and DISS. As shown in P < 0.01). The absorptive profiles of SA5 and DISS were close to that of the PRE incubated gut sacs over 180 min, while the absorption kinetic parameters such as Ka and Papp of SA6 cotreated with TS were higher (P < 0.01) than those of the PRE's. That is, when these three oligosaccharide esters were coincubated with TS, the cumulative amount per intestinal area of them was 1.59, 1.84, and 2.18 times more than that treated with monomers alone at 180 min. There were statistically significant differences in these measurements (P < 0.01). This showed that although the absorption of polygala saponins was poor , is the major compound of xanthones in PR. Polygala saponins have many pharmacological activities, including antidepression, neuroprotection, and antidementia –30. All  Figures , they maKa) of SA5 and SA6 were 0.044 ± 0.0077 μg·cm2·min (n = 5) and 0.066 ± 0.010 μg·cm2·min (n = 5) faster (P < 0.01) than that of non-verapamil-treated gut sacs. In 180 min, verapamil increased Papp from (1.34 ± 0.092) × 10−5 to (1.45 ± 0.093) × 10−5cm/s for SA5 and from (0.80 ± 0.079) × 10−5 to (1.27 ± 0.034) × 10−5cm/s for SA6 with a significant difference (p < 0.01). This indicated that the intestinal transportation and absorption of SA5 and SA6 could be markedly enhanced along with the inhibition of P-gp induced by verapamil and the bioavailability of SA5 and SA6 could subsequently be increased. However, there was no significant influence on 3,6′-disinapoylsucrose transport by verapamil, which suggested that DISS was not a substrate for P-gp. This is consistent with the results of Chen et al. [In addition to providing information on absorption mechanisms of drugs, the everted gut sac model was also useful to explore drug interactions with the ATP binding cassette transporter proteins including P-glycoprotein (P-gp), which is enriched in the small intestine. P-gp may reduce the absorption of P-gp substrate drugs. Inhibition of P-gp's function or decreasing its expression by use of a P-gp inhibitor like verapamil could improve the bioavailability of P-gp substrate drugs . In orden et al. .Papp of DISS, which means that DISS might be transported across the intestinal mucosa by paracellular passive penetration. In the present study, sodium caprate (1%) was coadministered with SA5 (0.050 mg/ml), SA6 (0.086 mg/ml), and DISS (0.198 mg/ml) to observe the possible influences on the intestinal absorption of the three oligosaccharide esters. The cumulative absorption amount of these esters per area increased up to about 1.87-, 2.08-, and 2.00-fold, respectively (P < 0.01), at 180 min by combination with SC (Ka) of the three oligosaccharide esters were 0.061 ± 0.0077 μg/cm2/min (n = 5), 0.0963 ± 0.010 μg·cm2·min (n = 5), and 0.069 ± 0.0031 μg·cm2·min (n = 5) faster, respectively (P < 0.01), than that of monomers alone. These results suggested that SC could significantly enhance and increase the absorption of SA5, SA6, and DISS in the small intestine. The increased absorption might indicate that the absorption mechanism of SA5 and SA6 combined active transport with paracellular passive penetration, while the absorption mechanism of DISS was dominated by paracellular passive penetration.The literature  states t with SC . In addiRho123 is a well-known substrate for P-gp, whose absorption can be significantly enhanced by P-gp inhibitors, such as verapamil. . Therefo in vivo, in vitro, and in situ routes. The in vitro everted rat gut sac model is a simple and efficient method to study the permeability and absorption kinetics of oral drugs to provide information on absorption mechanisms of drugs. Compared with other in vitro methods, it has an advantage for analytical experiments because the volume in the intestinal sac is small, and the concentration of a drug can be relatively high [Oral administration is the most common route for TCM. It is important to investigate the intestinal absorption process of the main ingredients in TCM. Up to the present, several intestinal absorption methods or models includeely high . In the In conclusion, by using the everted gut sac model, the overall absorption profile of SA5, SA6, and DISS can be enhanced by combination with TS, which are other effective components in PR. We showed that both the P-gp inhibitor verapamil and paracellular enhancers, such as sodium caprate, could significantly enhance the absorption of SA5 and SA6. This suggests that the absorption mechanism of SA5 and SA6 combines active transport with paracellular passive penetration. Therefore, the data from the Rho123 absorption experiment led us to conclude that TS can enhance the intestinal absorption of SA5 and SA6 like a P-gp inhibitor would. But this is not the whole reason. Indeed, the absorption of DISS was affected only by sodium caprate, and its absorption was dominated by paracellular passive penetration. Further research should be carried out to understand the relationship between polygala saponins and the absorption of SA5, SA6, and DISS by paracellular passive penetration.


Vibrio alginolyticus is one of the most important pathogens in mariculture and leading to heavy losses. After treatment with Cu2+, Pb2+, and low pH, the expression of oxidative phosphorylation pathway genes, including coxA, coxB, coxC, ccoN, ccoO, and ccoQ, was found commonly downregulated by RNA‐seq as well as quantitative real‐time PCR. RNAi significantly reduced the expression of coxA, coxB, coxC, ccoN, ccoO, and ccoQ in V. alginolyticus. Compared with the wild‐type strain, the adhesion abilities of RNAi strains of V. alginolyticus were significantly impaired, as well as their cytochrome C oxidase activity. ccoQ appeared to be more important in the regulation of bacterial adhesion in these target genes, while ccoO was relatively weak in the regulation of the adhesion. Meanwhile, the changes of temperature, salinity, pH, and starvation affected coxA, coxB, coxC, ccoN, ccoO, and ccoQ expression remarkably. These findings indicated that: the oxidative phosphorylation pathway is a critical regulator of adhesion in V. alginolyticus; coxA, coxB, coxC, ccoN, ccoO, and ccoQ regulate the bacterial adhesion in response to environmental changes such as temperature, salinity, pH, and starvation. The purpose of this study is to reveal the relationship between bacterial adhesion and oxidative phosphorylation pathway under adversities.22.1Vibrio alginolyticus high pathogenic strain ND‐01 was isolated from mariculture fish large yellow croaker  broth added with 2% NaCl at 28°C  , Pb (100 mg/L), and Cu (50 mg/L) were carried out on V. alginolyticus was grown at 4, 15, 28, 37, and 44°C, respectively, in LB broth    annotation of the unigenes, while WEGO was applied for GO functional classification analysis of all genes and understand the distribution of gene functions at the macro level. GO terms with Bonferroni corrected http://www.genome.jp/kegg/) was used for the KEGG pathways annotation. Q value ≤ 0.05 was taken as a threshold, while pathways fulfilling this were regarded as significantly enriched pathways in DEGs  were used for mucus collection via our previously described procedure  was used to extract the total RNA from the bacterial cells following the manufacturer's protocol. A Rever Aid Mu‐MLV cDNA synthesis kit (TransGen Biotech) was used to synthesize first‐strand complementary (c) DNA from 2.0 mg total RNA following the manufacturer's protocol  21–23 nt) with a characteristic and highly specific 2‐ to 3‐nucleotide 3′ overhang was obtained from GenePharma Co. Ltd. Sequences of siRNA used here are listed in Table –23 nt wiV. alginolyticus competent cells was transferred to the cuvette after being chilled on ice for 30 min, followed by electroporation  with a Bio‐Rad MicroPulser . After adding 900 μl of LB immediately, the mixture was incubated at 28°C for 1, 3, 6, 9, and 12 hr prior to RNA extraction, qRT‐PCR, and in vitro adhesion assay  sequences targeting their coding regions. The shRNAs were obtained from Shanghai Generay Biotech Co., Ltd.  was used to screen stable silenced clones . Relative Expression Software Tool (REST 2008‐version 2) was used to assess the relative expression levels of coxA, coxB, coxC, ccoN, ccoO, and ccoQ  was used to carry out qRT‐PCR with the SYBR green I fluorescent dye (Dongsheng) in accordance with the manufacturer's instructions  activity of 2.98 CFU/ml) was placed on the mucus‐coated glass slides, following by incubating humid at 28°C for 2 hr, and washing five times with PBS. The adherent bacteria were then fixed for 30 min by 4% methanol, dyed for 3 min by crystal violet, and counted with a microscope . Each group was conducted for five times, and 20 fields of view were selected randomly. Negative control was performed using PBS instead of bacterial suspension.According to the procedure described by Kong et al. , in vitr2.10SD and statistically analyzed with spss 18.0. Differences were determined by analysis of one‐way ANOVA followed by Dunnett's multiple comparison tests. A value of p < 0.05 was considered statistically significant.Results were showed as mean ± 33.12+, and Cu2+ could significantly reduce the adhesion ability of V. alginolyticus by 56.58%, 39.26%, and 37.41%, respectively. Therefore, RNA‐seq was performed on V. alginolyticus which was treated with low pH, Pb2+ and Cu2+.Results of previous study , subunit II of aa3‐type cytochrome c oxidase gene (coxB), subunit III of aa3‐type cytochrome c oxidase gene (coxC), subunit I of cbb3‐type cytochrome oxidase gene (ccoN), subunit II of cbb3‐type cytochrome oxidase gene (ccoO), and subunit IV of cbb3‐type cytochrome oxidase gene (ccoQ), respectively.There were six commonly downregulated DEGs in the oxidative phosphorylation pathway, and these DEGs were significantly changed in the Cups Figure, 2, 3. T3.2coxA, coxB, coxC, ccoN, ccoO, and ccoQ was carried out to validate the results of RNA‐seq. The results of qRT‐PCR matched those of the RNA‐seq: the low pH, Pb, and Cu treatments significantly reduced the expression of coxA , coxB , coxC , ccoN , ccoO , and ccoQ   siRNA treatments. The expression of these genes decreased significantly at 1–6 hr. After 6 hr, coxA and ccoQ were no longer changed significantly, while coxB, coxC, ccoN, and ccoO were remain significantly changed at 12 hr , coxB (by 4.20‐fold), coxC (by 2.77‐fold), ccoN (by 6.58‐fold), ccoO (by 7.08‐fold), and ccoQ (by 2.91‐fold) at 1 hr. Simultaneously, the adhesion of V. alginolyticus was significantly reduced by 1.97‐, 2.76‐, 2.28‐, 1.97‐, 2.32‐, and 3.02‐fold, respectively. Thus, ccoQ appeared to be more important in the regulation of bacterial adhesion in these target genes, while ccoO gene was relatively weak in the regulation of the adhesion.According to the results of data analysis, siRNA treatment significantly reduced 3.43.4.1coxA, coxB, coxC, ccoN, ccoO, and ccoQ were decreased significantly in stable silenced strains by 1.52‐, 2.10‐, 3.00‐, 1.85‐, 2.35‐, and 1.17‐fold, respectively. The expression level of coxC gene was the most impaired among six genes in the oxidative phosphorylation pathway.The results in Figure 3.4.2V. alginolyticus and stable silenced clones were shown in Figure coxA, coxB, coxC, ccoN, ccoO, and ccoQ‐RNAi V. alginolyticus were impaired by 77.80%， 65.72%， 44.40%， 49.20%， 50.80%, and 80.64%, respectively. The decrease of the adhesion ability of ccoQ‐RNAi V. alginolyticus was the largest among them, while the decrease of the adhesion ability of coxC‐RNAi V. alginolyticus was the least. These results indicated that stable gene silencing could significantly reduce the adhesion of V. alginolyticus.The adhesion ability of the control 3.4.3V. alginolyticus displayed well enzyme activity of CoxC and the stable silenced clones displayed defective enzyme activity of CoxC. The enzyme activity of CoxC of coxA, coxB, coxC, ccoN, ccoO, and ccoQ‐RNAi V. alginolyticus were impaired by 92.78%, 76.63%, 29.73%, 65.12%, 65.81%, and 95.70%, respectively. The enzyme activity of CoxC of coxA‐ and ccoQ‐RNAi V. alginolyticus were more impaired among them, and the coxC‐RNAi V. alginolyticus was less impaired. These results indicated that stable gene silencing of these six genes could significantly reduce the enzyme activity of CoxC.The results in Figure 3.5coxA, coxB, coxC, ccoN, ccoO, and ccoQ in different ways , coxC (by 1.18‐fold), ccoN (by 1.52‐fold), and ccoO (by 2.29‐fold) at 3 days, respectively. Therefore, among these genes, ccoO appeared to be the more sensitive gene to starvation.Although the expression levels of 4V. alginolyticus which were treated with Cu2+, Pb2+, and low pH, and these stress conditions reduced the adhesion ability of V. alginolyticus significantly  might be sensitive to environmental stresses. Previous studies have shown that starvation   , without a CuA center present, is a proton‐pumping terminal oxidase solely in bacteria , CcoO (subunit II), CcoP (subunit III), and CcoQ (subunit IV) proteins   . The results showed that the V. alginolyticus.In summary, the present study demonstrated that the oxidative phosphorylation pathway was sensitive to environmental factors, and it was related to the adherence of The authors declared that they have no conflicts of interest to this work.Q. Y. and Y. S. conceived and designed the experiments. L‐X. H., L. H., L. Z., and Y. Q. performed the experiments. Q. Y. and L‐X. H. drafted and revised the manuscript. All the authors approved the final version.All animal experiments were carried out strictly under the recommendations in the “Guide for the Care and Use of Laboratory Animals” set by the National Institutes of Health. The animal protocols were approved by the Animal Ethics Committee of Jimei University (Acceptance No. JMULAC201159).


Streptococcus thermophilus nutrient requirements to guide the design of media for high cell density culturing. The growth kinetics, physiological state, and nutrient requirement profiles of S. thermophilus were analyzed in chemically defined media. The results showed that the intracellular ATP concentration, H+‐ATPase activity, NADH/NAD+, and NH3 concentrations varied with intracellular pH. The nutrient components with the highest amounts required were Leu and Asp; ascorbic acid and p‐amino benzoic acid; K+ and PO43−; and guanine and uracil. The nutrient components with the largest required ratios were Arg, His, and Met; folic acid, cyanocobalamine, biotin, and nicotinic acid; Ca2+ and Mg2+; and guanine and uracil. In this study, different nutrient components were primarily used at different phase. Trp, Tyr, calcium pantothenate, thiamine, guanine, and Mg2+ were mainly used from late‐lag to midexponential phase. Met, Pro, Phe, Ala, Gly, nicotinic acid, and riboflavin were mainly used from midexponential to late‐exponential phase. The highest bioavailabilities of nutrient components were also found at diverse phase. Met, Leu, Ile, Asn, Glu, Lys, Pro, Gly, riboflavin, nicotinic acid, adenine, uracil, inosine, and Ca2+ had the highest bioavailability from late‐lag to midexponential phase. Lactose, Glu, Asp, His, Trp, Cys, Val, Arg, Phe, Ala, Ser, Thr, Tyr, folate and cobalamin, calcium pantothenate, ascorbic acid, thiamine, biotin, p‐amino benzoic acid, vitamin B6, K+, Mg2+, guanine, xanthine, and PO43− had the highest bioavailability from midexponential to late‐exponential phase. This study elucidated the nutrient requirement profiles with culture time and biomass at various average growth rates during the growth of S. thermophilus. The present results will help to formulate complex media for high cell density cultivation and provide the theoretical basis for S. thermophilus feeding strategies.This study aimed to evaluate the profiles of  The effect of specific nutrients on LAB proliferation is dependent on the nutrient requirements of a given species  displays excellent properties in cheese and yogurt starter. The goal of this study was to evaluate the amino acids, vitamins, ions, purines, pyrimidine, inosine, and lactose requirements for the growth of ST‐MZ‐2 by assessing average growth rates with respect to culture time and biomass yields to help design of a high cell density cultivation medium for ST‐MZ‐2 and study the metabolic mechanism of nutrients.Previous studies have shown that 22.1ST‐MZ‐2 was obtained from a previous study  from eighth subcultures. The pellet was washed twice with PBS buffer  and the culture was inoculated in a 10‐l Biotech‐7000 bioreactor  containing 7 L of CDM. The temperature and rotation speed were fixed at 42.5°C and 150 rpm, respectively. The pH was maintained at 6.25 by the automatic addition of 1 mol/L NaOH. The culture supernatant and cells were obtained by centrifugation  and were preserved at −80°C for further analysis. Batch fermentations were repeated five times. Cell extract preparations were performed according to the methods of Zhengwen et al. . The protein content of the cell‐free extracts was determined with a protein assay kit . NADH and NAD+ were quantified using HPLC with a ZORBAX SB‐Aq column  and a UV detector  of ST‐MZ‐2 was measured spectrophotometrically at 650 nm. The dry weight (DW) of cultured cells was determined gravimetrically. The intracellular pH was measured according to the method of Breeuwer, Drocourt, Rombouts, and Abee . The cul6, thiamin‐HCl, and cyanocobalamine concentrations were determined by RP‐HPLC with ultraviolet detector . Lactose was quantified by HPLC using a chromatographic system composed of an Alliance 2695 module injector , a 2414 Differential Refractometer detector (Waters), and an ICSep ION‐300 ion exchange column , all controlled with Empower software (Waters) described by Del et al. . The fol33.1+‐ATPase activity were highest at end‐lag phase and gradually increased from midexponential growth phase to stationary phase . The consumption amounts of other vitamins ranged from 1.6 to 36.1 nmol/L. The consumption amount of folic acid was 98.68% of the addition amount, which was highest, and thiamin‐HCl (48.49%) was the lowest. The consumption amounts of other vitamins ranged from 59.86% to 89.16% of the addition amounts. Folic acid, ascorbic acid, vitamin B6, biotin, and p‐amino benzoic acid exhibited the highest consumption at lag phase, whereas calcium pantothenate and thiamine exhibited the highest consumption at T2. Nicotinic acid and riboflavin had the highest consumption at T3. In Figure p‐amino benzoic acid was highest at stationary phase.In Figure amin‐HCl 8.49% was3.5In Figure 3.6+ (9.32 mmol/L), which was 17.71% of the addition amounts. The amount of PO43− consumed was 4.6 mmol/L, and was 13.49% of the addition amount, which was the lowest. The consumption amount of Mg2+ was 0.19 mmol/L and was 19.59% of the addition amount. The amount of Zn2+ consumed (3.4 nmol/L) was the lowest. Fe2+ exhibited a low consumption amount, whereas it was 46.44% of the addition amount, which was the highest. The amount of Mn2+ and Ca2+ consumption was 0.04 and 1.32 mmol/L, and was 24.66% and 29.53% of the addition amounts, respectively. Mn2+, Zn2+, Ca2+ and PO43− exhibited the highest consumption at lag phase, whereas Fe2+ and Mg2+ exhibited the highest consumption at T2. K+ had the highest consumption at stationary phase. In Figure 2+ and K+ showed similar consumption patterns from end‐lag phase to stationary phase. The consumption of PO43−, Mn2+, and Fe2+ decreased with increasing μ, indicating a more effective utilization of these ions at a higher μ with the increasing of biomass as the biomass yield based on consumption increased. In addition, Zn2+ and Ca2+ had similar consumption patterns from end‐lag phase to stationary phase. Their biomass utilization efficiencies were the highest at T2, whereas that of the other ions was highest at T3. All assayed ions, with the exception of Mg2+, exhibited the lowest biomass utilization efficiencies at lag phase and stationary phase.In Figure 4+‐ATPase to maintain the intracellular pH (Kilstrup, Hammer, Ruhdal, & Martinussen, + participates in glycolysis as the coenzyme of dehydrogenase, which brings a large amount of ATP to pump H+ out of the cells (Ningzi et al., +‐ATPase activity, NADH/NAD+ ratio, and NH3 concentration are highest during lag phase, contributing to the higher intracellular pH during lag phase. The intracellular pH dropped as the sodium lactate concentration increases. This could be attributed to the pH homeostasis by H+/lactate symport, which was less effective at high external lactate concentrations (Konings & Otto, 3, ATP, H+‐ATPase, NAD+, and NADH required the consumption of nutrients (Fernández & Zúñiga, S. thermophilus encounters two stress factors simultaneously during culturing, including the gradual increase in intracellular acidity and extracellular osmotic pressure (Shan et al., The intracellular pH of LAB is low when they are initially inoculated into new media but needs to be adjusted to a high intracellular pH during lag phase to grow better during exponential growth phase (Rault, Bouix, & Béal, S. thermophilus was observed to consume a large amount of Leu, Glu, Lys, Ala, Arg, and Asp in M17 medium (Guimont, LAB have multiple amino acid requirements to synthesize proteins, provide precursors for the biosynthesis of amino acids, nucleotides, and vitamins, generate metabolic energy, control intracellular pH, and resist stresses (Iyer, Tomar, Maheswari, & Singh, Guimont, , which wGuimont, . CompareGuimont, .S. thermophilus and exhibited the highest consumption at lag phase. The primary role of lactose may be to provide energy for the regulation of intracellular physiological states at lag phase. Vitamins are essential micronutrients that are typically used as precursors of various enzymes that are necessary for vital biochemical reactions in all living cells (LeBlanc et al., Lactobacillus sakei and L. plantarum. In addition, biotin was observed to be a stimulatory factor for the growth of some LAB strains or an essential factor for other strains (Hayek & Ibrahim, p‐amino benzoic acid, and the consumption rate of folic acid was the highest. Riboflavin, nicotinic acid, and calcium pantothenate were required by S. thermophilus and they promote the growth of bacteria (Letort & Juillard, Lactose provided energy and a carbon source for growth of L. lactis MG1363 in chemically defined medium (Martinussen, Wadskov‐Hansen, & Hammer, L. lactis (Ryssel et al., Nucleotides are substrates for RNA and DNA synthesis, constituents of coenzymes, are primary energy donors, and activate precursors in peptidoglycan and lipid synthesis (Kilstrup et al., 2+ and Ca2+is known to stimulate the growth and improve the survival of S. thermophilus (Letort & Juillard, 2+ and Mg2+, more so by Ca2+ (Tham et al., 2+ and Ca2+ was highest at T2 and T3, respectively. Their consumption exhibited a complementary trend at these two stages. Furthermore, ST‐MZ‐2 required Mg2+ and Ca2+ to stimulate its growth at T2 and T3. Mn2+ has been observed to enhance β‐glucosidase activity (Jeng et al., 2+ had the highest biomass utilization efficiency. The acid phosphatase and β‐glucosidase activity was inhibited by Fe2+ and Zn2+ (Jeng et al., 2+ and Zn2+ from CDM did not affect the growth of S. thermophilus (Letort & Juillard, MgThe ratios of nutrients are crucial for high cell density cultivation of LAB. Shortage of nutrients limits the growth of LAB (Lahtvee et al., S. thermophilus (Letort & Juillard, S. thermophilus (Hong et al., S. thermophilus for better growth. S. thermophilus has the capability to synthetize nonessential nutrients using other nutrients (Hayek & Ibrahim, The optimization of balanced amounts of nutrients is important in the design of ideal medium for the high cell density cultivation of LAB. More importantly, we believed that biosynthesis of inadequate or unnecessary nutrients could be avoided in LAB by optimization of balanced amounts of nutrients. Studies have shown that some amino acids, vitamins, and nucleic acid bases are unnecessary for the growth of It should be understood that none of the authors have any financial or scientific conflict of interest with regard to the research described in this manuscript.