Melastoma malabathricum Linn. is a perennial traditional medicine plants that grows abundantly throughout Asian countries. In this study, M. malabathricum Linn. leaf hot water crude extract with anticoagulant activity was purified through solid phase extraction cartridge and examined for the bioactive chemical constituents on blood coagulation reaction. The SPE purified fractions were, respectively, designated as F1, F2, F3, and F4, and each was subjected to the activated partial thromboplastin time (APTT) anticoagulant assay. Active anticoagulant fractions  were subjected to chemical characterisation evaluation. Besides, neutral sugar for carbohydrate part was also examined. F1, F2, and F3 were found to significantly prolong the anticoagulant activities in the following order, F1 > F2 > F3, in a dose dependent manner. In addition, carbohydrate, hexuronic acid, and polyphenolic moiety were measured for the active anticoagulant fractions . The characterisation of chemical constituents revealed that all these three fractions contained acidic polysaccharides  and polyphenolics. Hence, it was concluded that the presence of high hexuronic acids and polysaccharides, as well as polyphenolics in traditional medicinal plant, M. malabathricum, played a role in prolonging blood clotting in the intrinsic pathway. Melastoma malabathricum Linn. is a small shrub belonging to the Melastomataceae family [ M. malabathricum Linn. is known as “senduduk,” and this species is widely accepted as a traditional medicinal plant [e family . This eval plant . M. malabathricum Linn. plant has been reported to be used as a folklore medicine. Decoction from the leaves is the most favourable as it is believed to be able to treat diarrhoea and dysentery and heal wounds and other ailments [ M. malabathricum Linn. exhibited antidiarrheal [The wholeailments . Besidesiarrheal  as well iarrheal , 6. M. malabathricum Linn. hot water extract has an outstanding anticoagulant activity. To our best knowledge, however, there has been no report on the chemical constituents of M. malabathricum Linn.hot water extract, which exhibits anticoagulant activity. Previous literature has also proven that the anticoagulant activity of Lythrum salicaria and Porana volubilis is contributed by acidic polysaccharides and polyphenolic compounds [ M. malabathricum Linn. leaf contain similar chemical constituents which demonstrate similar anticoagulant activity that is probably better than other higher plants such as L. salicaria and P. volubilis. Thus, in continuum with the previous study, the hot water crude extract of M. malabathricum Linn. leaf was fractionated, and anticoagulant activities of fractionated fractions were evaluated. At the same time, carbohydrate, hexuronic acid, and polyphenolic moiety in the active anticoagulant fractions were also measured. Overall, this study revealed that the fractions of M. malabathricum Linn. contained high amounts of carbohydrate, hexuronic acid, and polyphenolic, which are responsible for the anticoagulant activity. Such a discovery is crucial information as it provides clues for potential treatment of blood coagulant disorders.Manicam et al.  have shoompounds , 9. In tStandard human normal pooled plasma, STA-PTT automated reagent, and 0.025 M calcium chloride were purchased from Diagnostica Stago , while sodium bicarbonate, sodium borohydride, gallic acid, carbazole, glucuronic acid, arabinose, galactose, mannose, rhamnose, xylose, dextran standards, trifluoroacetic acid (TFA), and heparin sodium salt were bought from Sigma-Aldrich . Phenol, acetonitrile, ammonium hydroxide, glacial acetic acid, acetic anhydride, dimethyl sulfoxide (DMSO), and Folin-Ciocalteu's reagent were purchased from Merck . Concentrated sulphuric acid and glucose were purchased from Fisher Scientific , and sodium borate de carbohydrate was purchased from Biobasic . M. malabathricum Linn. leaves were collected between September and October 2009 from Lebuh Silikon, Universiti Putra Malaysia, in Serdang, Selangor, Malaysia. The samples were identified by a botanist and a voucher specimen was deposited at the Herbarium Biodiversity Unit, Institute of Biosciences, Universiti Putra Malaysia, under the reference number SK 1717/09.Fresh matured M. malabathricum was performed according to the method described by Manicam et al. [ M. malabathricum leaves (500 g) were refluxed for 5 h at 100°C with 1 L deionised water. After 5 h, the hot water extract was filtered using Whatman Grade number 1 filter paper , concentrated into dryness, lyophilised, and stored at −20°C until further analysis. Next, lyophilised crude extract (0.5 g) was dissolved in 100% deionised water and loaded into 10 g of C18, Sep-Pak Cartridge . Four different ratios of water to acetonitrile  were passed through the cartridge. The analysts of interests for each fraction were collected, dried and lyophilized. Four fractions designated as F1, F2, F3, and F4 were obtained. Each fraction was reconstituted with distilled water at different concentrations for anticoagulant activity assay and chemical characterisation.The hot water extraction from the leaf ofm et al. . BrieflyIn vitro intrinsic anticoagulant pathway was measured by activated partial thromboplastin time (APTT). Standard human normal pooled plasma was spiked with equal volume of different concentrations of M. malabathricum crude extract and fractions . Meanwhile, STA-PTT automated reagent was reconstituted and subjected to APTT assay, together with 0.025 M calcium chloride. The APTT assay was analysed using STA Compact coagulation analyser  according to the manufacturer's protocol. The anticoagulant activity was measured by using a parallel standard curve based on the standard calibration curve for heparin sodium salt (140 IU/mg).−1 resolution. The 1H NMR spectra were obtained from 500 MHz Varian INOVA NMR spectrometer . All the 1H NMR samples were dissolved in deuterium oxide (D2O).Total carbohydrate content was determined by using a modified phenol-sulphuric acid assay . AbsorbaμL water and adding 20 μL allose as an internal standard. Next, 20 μL of 15 M ammonium hydroxide and 1 mL of 0.5 M sodium borohydride solution in dimethyl sulfoxide were added. The samples were then left overnight at 4°C after being incubated in 40°C water bath for 90 min. On the following day, the samples were prewarmed at 40°C for 5 min. Subsequently, 100 μL of glacial acetic acid, 200 μL of 1-methylimidazole, and 1 mL of acetic anhydride were added. The samples were incubated at 40°C for 10 min to allow completion of acetylation. Finally, 2.5 mL of water and 1 mL of dichloromethane were added and centrifuged at 1000 ×g for 10 min. The lower dichloromethane part was analysed using GC. Meanwhile, arabinose, glucose, galactose, mannose, rhamnose, and xylose were used as standards. Aditol hexa-acetates were analysed using Agilent 6890 gas chromatography  with BPX-70 column . Helium was used as gas carrier on column injection and flame ionisation detector was utilized as a detector. The temperature was programmed at 50°C and maintained for 30 s, increased to 170°C at 50°C/min and finally to 230°C at 2°C/min before it was maintained for 5 min. The inlet temperature was 250°C, with a flow rate of 1 mL/min.Neutral sugar compositions were identified and quantified through gas chromatography (GC) analysis. Polysaccharides were subjected to hydrolysation, reduction, and acetylation , 14. ActMolecular mass of active anticoagulant samples was determined through Jasco high performance liquid chromatography (HPLC LC-1500 series) with refractive index detector . Phenomenex polysep-GFC-P-4000  was used with the flow rate at 0.8 mL/min, while the mobile phase was deionised water. Dextran standards of various molecular masses from 5 kDa to 270 kDa were used as standards for calibration.n = 3) and analysis was performed using one-way analysis of variance (ANOVA), followed by post hoc Dunnett's multiple range tests to determine the means of significance from the control . Significant differences for all the data sets were measured and designated as *P < 0.05, **P < 0.01, and ***P < 0.001.Data are shown as mean ± S.E.M.  and 264.15 ± 2.34 s (P < 0.001) compared to the crude extract, which exhibited 252.51 ± 2.01 s (P < 0.001) and 255.56 ± 3.68 s (P < 0.001) at 500 μg/mL and 1 mg/mL, respectively. Similarly, F2 also showed a similar anticoagulant characteristic as F1. It significantly prolonged the blood clotting time at a concentration as low as 125 μg/mL. The prolonged clotting time at 1 mg/mL for F2 was 249.37 ± 3.45 s (P < 0.001). However, F3 was only able to significantly inhibit blood clotting at 1 mg/mL and 500 μg/mL, with 234.14 ± 0.99 s (P < 0.001) and 137.11 ± 0.58 s (P < 0.05), respectively.The dark brown material (lyophilised crude extract) was fractionated into four fractions with increasing solvent polarity. The fractions were labelled as F1, F2, F3, and F4. The anticoagulant activity of each fraction was determined by automated partial thromboplastin time (APTT) assay. This is in continuum with Manicam et al.'s report, which showed thatI, and V . Overall Lythrum salicaria (0.17 IU/mg) and Filipendula ulmaria (0.5 IU/mg) and were therefore reported as anticoagulant medicinal higher plants [The anticoagulant activity for all the fractions was subsequently measured using the standard calibration curve of unfractionated heparin (140 IU/mg). As summarised in r plants .The chemical characterisation was carried out to have a better understanding of the chemical compositions for active anticoagulant fractions . F1 was a dark brown gum, while F2 was a brown material, and F3 was a light brown material. The yields for F1, F2, and F3 were 54.17%, 14.97%, and 8.8% (w/w) of dry crude extract, respectively. M. malabathricum Linn. hot water extract which was fractionated with increasing solvent polarity had decreased carbohydrate compositions in the F1, F2, and F3. From the results, F1 with the most potent anticoagulant fraction yielded the highest compositions of carbohydrate and uronic acid contents. The occurrences of carbohydrate and uronic acid compositions of fractions were correlated with the anticoagulant activities. The anticoagulant activities of the fractions decreased as the carbohydrate and uronic acid contents decreased in the following sequence F1, F2, and F3.Generally, carbohydrates, phenolic acids, and uronic acids were presented and they served as the main constituents for F1, F2, and F3 . It is iOn the other hand, F3 with moderate amounts of carbohydrate and uronic acid contents compared to F1 and F2 also displayed significant anticoagulant activity. This might be due to the presence of high percentage of phenolic acids in F3. Phenolic acids together with carbohydrates and uronic acids had been suggested to play a role in anticoagulant activities by Pawlaczyk et al. , 18 and Besides, three types of neutral sugars were being identified from F1, F2, and F3 active anticoagulant fractions. These findings suggest that rhamnogalacturonan, homogalacturonan, and rhamnose hexose types of pectic polysaccharide were presented. However, these neutral sugars occurred in small amounts only .The HPLC molecular mass analysis of F1, F2, and F3 showed broad and incomplete separated peaks. These unresolved molecular mass distribution patterns are typical profiles for polysaccharide conjugates that comprise mixtures of similar building fragments . BesidesF1 was the first fraction purified from the SPE extraction, which had been eluted with 95% water and 5% acetonitrile. The highest yield was obtained for F1, with 54.17% (w/w) of dry crude extract. From the HPLC molecular mass determination analysis, three major not well-separated peaks were identified. The molecular masses were ~280.08 ± 3.57 kDa, ~168.60 ± 3.57 kDa, and ~82.05 ± 3.22 kDa. However, the distinct peak at a retention time of 12.35 min could not be identified by using the set of dextran standards (data not shown). Molecular masses of sample analytes were separated based on the size. Larger molecular mass of analyte was eluted out first, followed by analyte with smaller molecular mass. In addition, retention time for 5 kDa of dextran standard was 10.94 min, while retention time for 27 kDa was 8.20 min. Therefore, this distinct peak with retention time at 12.35 min was postulated to have a molecular mass less than 5 kDa. Besides, fractionation through high water compositions of solvent resulted in the highest amounts of carbohydrates and uronic acids, with 78 ± 2.29% and 31.71 ± 3.96%, respectively. In spite of these, the phenolic acids content was the least abundant in F2 and F3. The percentage for phenolic acids present in F1 was 4.50 ± 0.49%. Meanwhile, F1 contained 2.39 ± 0.11% of rhamnose and 2.39 ± 0.15% of glucose. These results indicated the presence of rhamnose-hexose type polysaccharide.F2 was fractionated with 90% of water and 10% of acetonitrile containing 14.97% of the total mass. The molecular mass determination analysis for F2 revealed that F2 contains higher molecular mass components than F1. The average molecular mass of two dominant peaks was about ~267.48 ± 0.66 kDa and ~177.36 ± 1.63 kDa. Another distinguishable shoulder tailing peak was about ~224.85 ± 1.28 kDa. The presence of carbohydrates, phenolic acids, and uronic acids pointed out that F2 was a polyphenolic glycoconjugate. There were 46.34 ± 3.86% of carbohydrates, 34.76 ± 0.53% of phenolic acids, and 22.85 ± 3.60% of uronic acids found in F2. The neutral sugars compositions for F2 contained rhamnogalacturonan, homogalacturonan, and rhamnose-hexose types of pectic polysaccharides. The most dominant neutral sugar presence in F2 was rhamnose which was 6.01 ± 0.27%.F3 was purified with higher compositions of nonpolar solvent, with 80% of water and 20% of acetonitrile, and yielded 8.8% of the total dry weight of the crude extract. The HPLC molecular mass analyses for both F2 and F3 were alike. F3 also contained two dominants and one shoulder tailing peaks with ~271.24 ± 0.39 kDa, ~240.23 ± 0.04 kDa, and 183.49 ± 0.31 kDa, respectively. Phenolic acids were most likely distributed and extracted by higher compositions of nonpolar solvent. Therefore, F3 was rich in phenolic acids compared to F1 and F2. There was 65.12 ± 0.78% of phenolic acids found in F3. On the other hand, the carbohydrates and uronic acids compositions were the least abundant in F3, with 14.98 ± 1.96% and 15.42 ± 2.37%, respectively. Besides, there was only one neutral sugar-galactose found in F3. The percentage of galactose for F3 was 7.05 ± 0.09%, and type of pectic polysaccharide was suggested as homogalacturonan.−1. This spectral shape was identified as a stretching vibration of intramolecular hydrogen on hydroxyl (OH) group in carbohydrate [−1. These regions corresponded to the stretch vibration of aliphatic C–H groups derived from the methyl (CH3) group [−1. Stretching of phenolics C–C for F1, F2, and F3 was observed at the bands at 1470–1437 cm−1. In addition, the spectra band at 1396–1249 cm−1 indicated the presence of phenolic acids in the fractions. These bands were signature bands for phenolics C–O (H) stretching [The FT-IR spectrum of each active anticoagulant fraction  demonstrohydrate , 21. Bes3) group , 22. In 3) group  at 1771–−1, 2929 cm−1, and 2855 cm−1. The band spectrum at 3782 cm−1 was a signature band for carbohydrates. Carbohydrates were also abundantly found at strong signal bands of 1279–1199 cm−1. These bands could be due to the ring vibration of carbohydrates overlapping with stretching vibration of OH group and glycosidic vibration [−1 and 2855 cm−1 showed the stretching vibration of aliphatic carbon and hydrogen (C–H). The intensive band at 1470 cm−1 presented the characteristic of carboxylic functional group for uronic acids or phenolic acids.The FT-IR spectrum of F1 further proved that F1 did contain high amounts of carbohydrates and uronic acids. As shown in ibration . The ban−1. This band corresponded to the interaction of hydroxyl (O–H) group of carbohydrates and phenolic acids. Meanwhile, the intensive signals at 1699–1603 cm−1 demonstrated the stretching vibration of carboxylic group of uronic acids or phenolic acids in asymmetric and symmetric modes. Typical galacturonopyranose backbone of pectic polysaccharides was also presented at region 1181 cm−1.The characteristic of carbohydrates, phenolic acids, and uronic acids was displayed at F2 FT-IR spectrum. Broad band signal was exhibited at region 3361 cm−1. This region matched the characteristic of uronic acids or phenolic acids. Signals at region 1285–1249 cm−1 matched the FT-IR spectrum for phenolic acids. Moreover, spectra bands at 1044–1011 cm−1 displayed the stretching vibration of C–O for phenolic acids [F3, with the least abundance of carbohydrate moiety, did not have intense signal bands at the carbohydrate regions. Nevertheless, F3 did display a distinct band at 1566 cmic acids .1H NMR spectra for active anticoagulant fractions  also revealed the presence of carbohydrates and phenolic acids. Overall, the 1H NMR spectra were shown to be complex and displayed broad hump signals. These were due to the complex nature of F1, F2, and F3. The not well-resolved spectra were also caused by the overlapping of many chemical constituents within the samples. From δ 5.30–3.0 ppm.In agreement with the general chemical characteristics and FT-IR spectra, the 1H NMR spectrum for F1 was slightly different from that of 1H NMR of F2 and F3. F1, which was rich in carbohydrate contents, only displayed a complex signal at δ 5.30–3.0 ppm. This suggests the presence of C2–C6 atoms attached at the carbohydrate pyranose ring [δ 8.0–6.5 ppm. Both consisted of higher phenolic acids contents compared to F1. Furthermore, with the highest amount of phenolic acids, F3 was translated into having a more complicated signal than F2. The additional 1H NMR signals at δ 2.1–1.9 observed for F2 and F3 could be due to the substitution of acetyl, ethyl, or methyl groups for carbohydrates [The ose ring . Other thydrates . M. malabathricum Linn. fractions had high polydispersity of macromolecules. The active principles of the anticoagulant activity of these fractions were acidic polysaccharides and polyphenolics. This result further proved that the anticoagulant activities for M. malabathricum Linn. fractions were high when the polysaccharide and hexuronic acids contents were high. A plausible explanation for the anticoagulant activities is the presence of carboxylic acid from the uronic acids, which provided negative charge properties to the fractions [ Lythrum salicaria, Erigeron canadensis, and Porana volubilis have been found to contain polyphenolic polysaccharide conjugates as negative charged macromolecules for anticoagulant activity [Generally, F1, F2, and F3 showed significant prolonged blood clotting time, especially in the intrinsic pathway. The anticoagulant activities of the fractions were in an increasing manner of F1 > F2 > F3. These fractions were purer and they gave better anticoagulant activity profile compared to the crude extract. Taken altogether, and with all the chemical analyses, the results showed that active anticoagulantractions . Hence, activity , 18. Eveactivity , Guglielactivity , and Donactivity  reported M. malabathricum Linn. hot water leaf extract. This discovery opens up the avenue that the simple preparation of fractions and the abundant availability of M. malabathricum Linn. plant materials could lead to the development of M. malabathricum Linn. active anticoagulant fractions as safe and cheap natural oral anticoagulant agents. A further study to isolate single anticoagulant bioactive compound from active anticoagulant SPE purified fractions  and an investigation into the action of single bioactive compound in blood coagulation pathway can be carried out.All in all, the presence of negative charged polyphenolic-polysaccharides were suggested to have played a role in the anticoagulant activity, especially prolonging blood coagulation in the intrinsic pathway, as exhibited by the purified


During 1999–2010, a total of 49,762 deaths from drowning occurred in the United States, an average of 4,147 deaths per year. The average annual death rate from drowning for males  was more than three times that for females (0.7). The death rate for males was highest among those aged 1–4 years and ≥85 years . For females, the highest rates were among those aged 1–4 years (2.2) and <1 year (1.8).Source: National Vital Statistics System. Mortality public use data files, 1999–2010. Available at http://www.cdc.gov/nchs/data_access/vitalstatsonline.htm.


The aim of this study was to find out the ideal cut-off level of phosphate for safe healing when deformity correction and concomitant lengthening are indicated in the two different skeletal maturity groups of patients with rickets. Thirty-nine hypophosphatemic rickets patients were selected for the study and were divided into two groups: 27 skeletally immature (group IM) and 12 skeletally mature (group M). The outcomes were evaluated with respect to the healing index (HI), laboratory findings, and complications with the mean follow-up of 5.1 years . The healing index (HI) of group IM was 1.44 month/cm and HI of group M was 1.68 month/cm. The negative correlation between the level of serum phosphate and HI in group M (coefficient = −0.94) was evaluated to be less than the correlation in group IM (coefficient = −0.50), indicating that the HI is more likely to be affected by serum phosphate in group M than in group IM. Preoperative serum phosphate levels of 2.3 mg/dL and 2.6 mg/dL were analyzed to be the cut-off values of group IM and group M, respectively, in which the cut-off points divided the series into two groups having the most significantly different HI. Rickets is a group of diseases characterized by deficient or impaired mechanism of vitamin D due to a variety of etiologies. One of the most crucial clinical features is hypophosphatemia, which leads to poor bone mineralization and consolidation, resulting in limb deformities and short stature starting at young age. Low level of phosphate impairs phosphate-dependent apoptosis of hypertrophic chondrocytes and is considered the molecular basis for rickets. Phosphate plays an important role in the skeleton that extends beyond mineralized matrix formation and growth plate maturation and is critical for endochondral bone repair .Previous authors reported the surgical outcomes of deformity correction and variable affecting factors in hypophosphatemic rickets –4. And oSo we hypothesized that the optimal serum phosphate level for deformity correction and concomitant leg lengthening would be different for skeletally immature and mature patients and aimed to find the ideal cut-off level of phosphate for each group of skeletal maturity.With the approval of the review board of our institute, we retrospectively reviewed the history of a total of 61 patients who were diagnosed with hypophosphatemic rickets between the years 2002 and 2008. Diagnosis was made by the presence of radiologic findings, such as long bone cupping, splaying, and fraying of the metaphyseal areas and lab values such as elevated alkaline phosphatase level, low 25-OH vitamin D, and low percent tubular reabsorption of phosphate (TRP) . Among tUpon inclusion and exclusion of patients with respect to the criteria mentioned above, 39 patients were selected for this study. The patients were then divided into two main groups according to the physeal closure patterns of knee radiographs—27 skeletally immature (group IM) and 12 skeletally mature (group M) patients. The mean ages of the patients in group IM and group M were 9.6 years range: 3.8–15.8) and 23 years (range: 18.3–31.2) at the time of surgical treatment, respectively. The mean durations of follow-up of group IM and group M after initial treatments were 5.4 years (range: 3.1–7.6) and 4.8 years (range: 3.6–7.9). Only one side of the tibia of a patient was randomly selected and considered as one sample segment to minimize any bias caused by correlation of bilateral tibia of a patient [–15.8 andCorrective osteotomies were performed at the single, double CORA levels. The Ilizarov ring fixator was used for gradual or acute correction in all tibial deformities. The decision of performing concomitant leg lengthening with deformity correction was made after a thorough discussion with the patients and their families. Distraction was started 7 days postoperatively at a rate of 0.25 mm four times a day until the normalization of the mechanical axis and the desired length was achieved. The distraction rate was controlled according to the degree of callus formation. The external fixator was removed upon visualization of callus formation in 3 cortices at the distraction site , 10.μg/kg per day) was stopped about a week before surgery and was readministered when mobilization of the leg became possible, usually 5 to 7 days after the operation [The skeletally immature patients had been under medical treatment of hypophosphatemic rickets, which included anhydrous phosphate (Joulie's) oral solution  along with vitamin D or its metabolites. Joulie's solution was omitted in the morning of operation and was readministered as soon as bowl function returned after the operation. 1, 25-Dihydroxyvitamin D  and mechanical axis deviation (MAD) were measured using the radiographic images that were taken before the operation, at the time of removal of the external fixator and at the final follow-up. Additionally, the knee joint alignment was assessed using Fraser et al.'s criteria  and clasThe laboratory values of calcium, phosphate, and alkaline phosphatase (ALP) were assessed for biochemical evaluation.Complications were evaluated and classified into major and minor according to Paley's classification. The major complication did interfere with the original goals of treatment and the minor did not . Also thU test was used for data from two groups. The correlations between HI and other factors such as age and serum phosphate level were analyzed using linear regression model. The receiver operating characteristic (ROC) curve analysis was performed to determine the cut-off point in preoperative serum phosphate level by which the lengthened bone segments could be divided into two groups having the most significantly different HI.All statistical analyses were performed using SPSS . The Mann-Whitney In group IM, 24 patients had bilateral genu varum deformity and 3 patients had bilateral genu valgum deformity. In patients with genu varum deformity, the mean TFA averaged 20° (range: 2° to 36°) prior to the operation, 5° (range: 2° to 19°) at the time of removal of external fixator, and 6° (range: −8° to 22°) at the time of final follow-up. Among these patients, MAD averaged 3.5 cm (range: 4.4 to 5.8), 0.9 cm (range: −0.6 to 2.5), and 1.7 cm (range: −1.5 to 4.2) of the center of the knee joint preoperatively, at the time of external fixator removal, and at the time of final follow-up. Among the 3 patients who had bilateral genu valgum deformity in the IM group, TFA averaged −10° (range: −12° to −9°), −4.° (range: −11° to 1°), and 0° (range: −4° to 2°), and MAD averaged −16.0 cm (range: −20.0 to −14.0), −4.8 cm (range: −17.6 to 7.8), and 1.9 cm (range: −6.4 to 6) preoperatively, at the time of external fixator removal, and at the time of final follow-up.In group M, 10 patients showed genu varum deformity and 2 patients showed genu valgum deformity. Among patients with genu varum deformity, the TFA averaged 16° (range: 1° to 26°), 6° (range: −5° to 12°), and 6° (range: −5 to 17), and MAD averaged 4.0 cm (range: 0.4 to 8.3), 1.7 cm (range: −0.4 to 4.8), and 1.3 cm (range: −1.8 to 2.8) preoperatively, at the time of external fixator removal, and at the time of final follow-up. In two patients with bilateral genu valgum deformity, the TFA averaged −9° (−12° and −5°), 3° (−3° and 8°), and 1° (−5 ° and 6 °), and MAD averaged −2.1 cm (−3.0 and −1.2), 0.2 cm (−0.6 and 1.0), and −0.2 cm (−0.4 and 0.0) preoperatively, at the time of external fixator removal, and at the time of final follow-up .P = 0.04). The mean amounts of the lengthening were 4.43 cm (range: 1.70 to 10.46) and 4.96 cm (range: 2.14 to 8.33), respectively, without statistically significant difference (P = 0.56)  and 1.68 months/cm (range: 0.85 to 2.57), respectively, yielding statistically significant difference ( = 0.56) .P = 0.01), and the mean ALP levels were 574.96 U/IL (range: 352–1022) and 149.50 U/IL (range: 85–265), respectively, (P = 0.01). The mean levels of calcium were 9.32 mg/dL (range: 8.73–9.82) and 10.03 mg/dL (range: 9.14–13.14) and showed no statistical difference (P = 0.25)  and 2.21 mg/dL (range: 1.73–2.84) in group IM and group M ( = 0.25) .P = 0.11) (P = 0.86).The correlation between healing index and age of the patients was evaluated. Even though the relationship between HI and age showed a linear pattern, there was no statistically significant correlation between the two parameters in linear regression model ( = 0.11) . Also, tγ = −0.72, P = 0.01). Then the correlation of the parameters in groups IM and M was analyzed separately. The correlation equation and linear patterns in The correlation between HI and preoperative serum phosphate level of the patients was shown . There wP = 0.04). In group IM, the HI of those with a serum phosphate level higher than 2.3 mg/dL averaged 1.52 months/cm, while those with a serum phosphate level below 2.3 mg/dL averaged 1.82 months/cm. They were also significantly different (P = 0.03). Other biochemical parameters did not show any significant correlations with HI.The ROC curve analysis provided the optimal cut-off value for the serum phosphate level as being 2.6 mg/dL in group M and 2.3 mg/dL in group IM. In group M, the HI of those with a serum phosphate level higher than 2.6 mg/dL averaged 1.35 months/cm, while those with a serum phosphate level below 2.6 mg/dL averaged 1.74 months/cm. They were significantly different . However, there were no statistically significant differences in all other complications in all subgroups.In group IM, there were 26 major complications and 22 minor complications in the 15 patients whose serum phosphate level was below 2.3 mg/dL and 15 major and 17 minor complications among the 12 patients whose serum phosphate level was above 2.3 mg/dL. In group M, there were 11 major and 9 minor complications among patients whose phosphate level was below 2.6 mg/dL and 3 major and 7 minor complications observed in patients who had a higher serum phosphate level than 2.6 mg/dL. Recurrent deformity, leg length discrepancy, refractures, and delayed union were the common bony complications . The numHypophosphatemic rickets is a disorder characterized by markedly low serum phosphate level and abnormal mineralization of bone. The defective bone mineralization results in rachitic changes at the growth plate and osteomalacia in trabecular and cortical bones . The traPrevious authors reported various treatment modalities of deformity correction with or without limb lengthening in hypophosphatemic rickets patients , 10, 14 β superfamily. BMPs have been shown to play important roles in embryonic organ development, limb formation, and fracture healing including those that exhibit nonunion [The study of Wigner et al.  reportednonunion , 16–18. nonunion . Althougnonunion  performeThere were several limitations to this study. Firstly, this series involved a retrospective study of patients only whose tibia was osteotomized monofocally or bifocally. Many other patients who underwent surgeries for trifocal tibial corrective osteotomy or both tibial and femoral deformity corrections were excluded from the study. Thus, the cut-off serum phosphate level may not be representative of the entire population of hypophosphatemic rickets patients. However, gradual correction and lengthening were applied at the proximal osteotomy site and acute correction was applied at the distal osteotomy site in the bifocal deformity correction procedure. Healing index was assessed in reference to the result of gradual correction and lengthening applied at the proximal osteotomy site. So we believe the bias reflected on our result should be minimal. Secondly, other operative modalities such as intramedullary rod nailing and plate fixation were concomitantly used in a number of patients to prevent complications during or after deformity correction, hence causing bias in healing index.In conclusion, this study demonstrated the ideal cut-off level of serum phosphate for safe healing after deformity correction and concomitant limb lengthening procedures in both skeletally mature and immature patients with hypophosphatemic rickets. And it should be carefully taken into consideration preoperatively according to the patient's skeletal maturity. If the patient's serum phosphate level does not reach the cut-off level but is willing to undergo deformity correction or leg lengthening, we suggest that the patient may try phosphate supplementation until his/her serum phosphate level is maintained above the cut-off level for prevention of any undesired complications and delayed union. We believe our findings may suggest more cautious guidelines for meticulous workup plans towards safe healing during preoperative planning and consultation with the patients and their guardians.


Delonix regia and Cassia fistula seed extracts were evaluated for their antioxidant activity, total phenolics, ash, zinc and fatty acid content. Fourier Transform Infrared Spectroscopy (FTIR) was utilized to assess the chemical functionalities present within the seeds. Antioxidant activity was determined by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Trolox equivalent antioxidant capacity (TEAC) assays. Total phenolics were determined by the Folin-Ciocalteu assay. Lipid extracts were characterized by nuclear magnetic resonance spectroscopy and gas chromatography/mass spectrometry. Zinc concentration was determined by atomic absorption spectroscopy. Extracts from the seeds of C. fistula had a higher antioxidant activity, free radical scavenging activity, and phenolic content than D. regia. FTIR revealed that the seeds are a rich source of protein with small quantities of fat. C. fistula extracts contained a higher percentage of total fat than D. regia. Palmitic acid was identified as the predominant saturated fatty acid in both extracts. Oleic acid and linoleic acid were identified in smaller quantities. Seed extracts may be considered for use in food and nutraceutical applications. Delonix regia and Cassia fistula can be found interspersed throughout the island of Jamaica and are primarily ornamental in nature. There is limited knowledge regarding the chemical composition of the seeds from trees grown in Jamaica. D. regia, Raffin  also known as “flamboyant”, or “flame tree”, belongs to the Caesalpiniaceae family . The tree is known to reach heights of approximately 12 m, whereas the flowers, which cover the tree crown, show colours ranging from orange to red. Carotenoids  and anthocyanins  are responsible for the vibrant colours observed in the petals of the flower [ D. regia flower extract is known to also contain flavonols such as quercetin and its glycosides , as well as kaempferol rhamnosylhexoside and isorhamnetin rhamnosylhexoside [ Cassia fistula L.  is a semiwild Indian Laburnum that is widely cultivated in Mauritius but can also be found in Asia, Africa, Latin America, and the Caribbean. The tree has yellow flowers and is sometimes referred to as Golden Shower. C. fistula has been identified as a potentially novel source of free radical scavenging compounds [D. regia and C. fistula seed pods were randomly collected from trees located on the campus of the University of the West Indies, Kingston, Jamaica. Seeds were removed from mature dry pods. Ten pods represented one sample subset. Whole seeds were ground using a laboratory mill .A1 is absorbance of sample at 517 nm and  A0 is absorbance of control at 517 nm.The DPPH assay was performed according to the method of Brand-Williams et al. . SamplesμL) reacted with Folin-Ciocalteu reagent  and mixed for 5 min followed by addition of Na2HCO3 solution . The solution was incubated at 22°C (1.5 h) and the absorbance was measured at 760 nm using a spectrophotometer . A standard calibration curve of gallic acid (0–200 mg/L) was generated and the results were expressed as mg gallic acid/g.Total phenolics were determined using the Folin-Ciocalteu assay with modifications . SamplesμL) were combined with ABTS• (2 mL) and the absorbance readings were recorded after 2 min at 750 nm. A standard calibration curve of Trolox (0–300 μM) was prepared.Free radical scavenging activity of methanolic extracts was also determined as Trolox equivalence . 2,2′-Az in vacuo. Percent crude fat (dry weight basis) was determined gravimetrically.Oil was Soxhlet extracted from the dried, milled seeds with petroleum ether , and concentratedμL) were transmethylated with methanol/acetyl chloride solution [Soxhlet extracted oil samples (50 solution . The resμL) was chromatographed on an HP6890 series Gas Chromatograph interfaced with an HP5973 Mass Selective Detector. Constituent FAMEs were eluted with helium carrier gas (flow rate 1 cm3/min) through a DB-VRX column  in an oven programmed at 60°C for 3 min and increased at a ramp rate of 10°C/min up to 250°C for 15 min. Samples were injected at 230°C while the detector was maintained at 250°C. Constituents were identified by matching the mass spectra, National Institute of Standards and Technology (NIST) library of mass spectra .Methylated oil in hexane  at 500 MHz. A 5 mm probe was used for 1H NMR and 13C NMR experiments. Lipid extracts were run in deuterated chloroform (CDCl3) at 25°C, with tetramethylsilane as the internal standard while for phenolic extracts, deuterated acetone was utilized as solvent.x, y, and z are coefficients  [Iodine values were calculated based on the FAME content and were calculated utilizing the formula: = 2.62) .−1. Stock solutions of zinc were made and standard calibration curves were prepared. Results are expressed as ppm.Samples (1.5 g) were ashed in a muffle furnace (600°C for 1.5 h) and the resulting ash dissolved in HCl , diluted with deionized water, and filtered. Measurements for zinc were made using a Perkin Elmer 2380 Flame Atomic Absorption Spectrophotometer system equipped with the corresponding hollow cathode lamp at the time of analysis. The following parameters were utilized: lamp current 10 mA, wavelength 214 nm, and slit width 0.7 nm, with flame type consisting of air/acetylene and stoichiometric fuel flow at 0.9 to 1.21 min D. regia and C. fistula. OPUS software was used to acquire and manipulate the spectral data.A Bruker Vector 22 Fourier Transform Infrared (FTIR) Spectrometer was utilized to record the infrared spectra of milled seed samples ofSamples were analyzed in triplicate. Means and standard deviations of the data were presented. D. regia and C. fistula continues to increase with recent papers documenting their potential health benefit and applicability. Galactomannan, a storage polysaccharide, was isolated from the seeds of D. regia [ C. fistula [ C. fistula has shown antifeedant and larvicidal effects on insects. Rhein  was identified as the active component from the ethyl acetate extract [Interest in the chemical and physiological properties of extracts fromD. regia  with potD. regia . Sesquit fistula . The eth extract .Lens culinaris) and faba beans (Vicia faba minor) possessed high levels of antioxidant activity [ p-hydroxybenzoic acid [Legumes are a source of natural antioxidants . Their aactivity . The domoic acid . C. fistula have been used extensively in traditional Indian medicine. High levels of antioxidants and phenolics (proanthocyanidins and flavonoids) have been detected in the pods [ D. regia also possess antioxidant and antimicrobial attributes [ D. regia leaf extracts exhibit anti-inflammatory activity [ D. regia has been previously reported [Extracts fromthe pods  with barthe pods . Leaf, ftributes . D. regiactivity . Whereasreported , there i D. regia and C. fistula seed extracts was determined by the DPPH free radical scavenging and TEAC assays and total phenolics by the Folin-Ciocalteu assay. The DPPH assay is frequently utilized in assessing the antioxidant properties of extracts [The antioxidant activity ofextracts . The DPP C. fistula compared to D. regia  compared to D. regia (1.41 ± 0.65%). Palmitic acid was identified as the predominant fatty acid in both seed extracts  and benzyl alcohol (3.27 ± 0.48%) were also identified in seed extracts from C. fistula. Increased oil yields may be obtained by the utilization of a pressure based system for extraction. The oil extract of cowpeas (Vigna unguiculata) has been shown to possess antidiabetic properties [ C. fistula and D. regia may be evaluated for potential medicinal activity.extracts . Prior s present –26. LinoD. regia . The prelcohol 3. ± 0.48% 1H NMR data  and a multiplet (δ 5.26) which are due to the protons on the triacylglycerol backbone  and δ 2.70 (C. fistula) which are indicative of protons on a bis allylic carbon [α) and the 2-glycerol carbon (β) were observed at δ 172.65 and δ 173.09, respectively, in D. regia and δ 172.88 and δ 173.33 in C. fistula (δ 128–130 ppm in the 13C NMR (1H NMR spectroscopic data of methanolic extracts showed evidence of a methyl group (δ 1.36), ether linkages (δ 3.63–δ 3.81), phenolics (δ 4.12), and vinylic protons (δ 5.35 and δ 5.42). The presence of the methyl and ether functionalities suggests that the phenolic compounds present are substituted. The vinylic protons resonated as doublets and are due to the aromatic ring of the phenolics. The 13C NMR spectra also confirmed the presence of vinylic carbons (δ 129.07 and δ 130.93). Carbons with hydroxyl functionalities were observed at δ 62.19, δ 63.38, δ 64.03, and δ 64.25. NMR spectroscopy is increasingly being utilized to determine sample authenticity [NMR data  of the lbackbone . Olefinic carbon  due to t fistula , Table 5enticity . C. fistula and D. regia was similar (−1 (D. regia) and 3285.75 cm−1 (C. fistula) are indicative of N-H stretching vibrations present in the amide functionality of proteins. Amide N-H bending vibrations were observed at 1542.60 cm−1 (D. regia) and 1539.80 cm−1 (C. fistula). For secondary amides, the N-H bending vibrations were observed at 1238.54 cm−1 and 1243.64 cm−1 [−1, 1402.21 cm−1, 1645.96 cm−1, and 1644.30 cm−1 were due to the carbonyl functionality of the amides. The appearance of these strong bands indicates the presence of protein in the solid state [−1 (C. fistula) and 1456.76 cm−1 (D. regia) were due to bending deformation of C-H vibration with stretching vibrations being observed at 2852.90 cm−1, 2922.57 cm−1, 2857.90 cm−1, and 2926.49 cm−1. Other pronounced bands were observed at 1742.63 cm−1 (D. regia) and 1745.18 cm−1 (C. fistula) due to the carbonyl stretching vibrations of triacylglycerols present [−1 (D. regia) and 3363.25 cm−1 (C. fistula) which is due to the hydroxyl functionality of the phenolics present (−1 (C. fistula) and 1720.19 cm−1 are indicative of the presence of a carbonyl substituent on the phenolic ring. Sharp bands at 1052.94 cm−1 (C. fistula) and 1049.09 cm−1 (D. regia) also indicate that the phenolics present may be further substituted producing the carbon-oxygen stretching vibration  and 5.48 ± 0.05% (C. fistula) ash. Zinc is an essential trace element which has antioxidant [ C. fistula and D. regia seed extracts were found to contain similar concentrations of zinc, 34.5 ± 0.3 ppm and 38.4 ± 1.0 ppm, respectively.Seed extracts were found to contain 3.83 ± 0.60% (ioxidant  and antiioxidant . The abiioxidant . Seed sa D. regia and C. fistula seeds was investigated in the form of powder, ash, oil, phenolics, and antioxidant activity. The results indicate that the seeds are a rich source of proteins with fats and starch being present in small quantities. The seeds are also a source of antioxidants. This study further substantiates that extracts from the seeds of these trees may be of commercial value and warrant further investigation.The composition of


Background. Cancer is a significant disease burden in the world. Many studies showed that heavy metals or their compounds had connection with cancer. But the data conflicting about the relationship of manganese (Mn) to cancer are not enough. In this paper, the relationship was discussed between Mn concentrations in drinking water for rural residents and incidence and mortality caused by malignant tumors in Huai'an city. Methods. A total of 158 water samples from 28 villages of 14 towns were, respectively, collected during periods of high flow and low flow in 3 counties of Huai'an city, along Chinese Huai'he River. The samples of deep groundwater, shallow groundwater, and surface water were simultaneously collected in all selected villages. Mn concentrations in all water samples were determined by inductively coupled plasma-mass spectrometry (ICP-MS 7500a). The correlation analysis was used to study the relationship between the Mn concentration and cancer incidence and mortality. Results. Mn concentrations detectable rate was 100% in all water samples. The mean concentration was 452.32 μg/L ± 507.76 μg/L. There was significant difference between the high flow period and low flow period  and also among deep groundwater, shallow groundwater, and surface water . The ratio of superscale of Mn was 75.32%. There was significant difference of Mn level between samples in the high flow period and low flow period  and also among deep groundwater, shallow groundwater, and surface water . And also we found that, during the low flow period, Mn concentration has positive correlation with cancer incidence and mortality; for a 1 μg/L increase in Mn concentration, there was a corresponding increase of 0.45/100000 new cancer cases and 0.35/100000 cancer deaths (P < 0.05). Conclusions. In Huai'an city, the mean concentration of Mn in drinking water was very high. Mn concentration correlated with cancer incidence and mortality. Based on the GLOBOCAN 2008 estimates, about 12.7 million cancer cases and 7.6 million cancer deaths are estimated to have occurred in 2008 [0.54/105 . Huai'anManganese (Mn) is widely distributed as one of the most abundant elements in the earth's crust and also in many foodstuffs. Mn is an essential trace element with many biological functions but toxic at higher doses , 7. In iHuai'an city is located in the center of Jiangsu province across Huaihe River. It has 5.4 millionpeople and 70% of them are rural residents. It has four counties, four districts, and one economic development zone which contain in all 116 towns. Here it has a high cancer incidence especially digest system neoplasm. Currently, only Huai'an city and its urban district use central water supply, while the other towns use small central water supply in rural areas. Before quality improvement, drinking water for rural residents mainly came from well and hand pressure well or earlier from the ditch and pond.3 was added to keep pH < 2. Then the samples were transported to the lab and were stored under 4°C. The water samples were analyzed by ICP-MS 7500a.We chose 5 towns from Jinhu County and Chuzhou county, respectively, and 4 towns from Xuyi County and chose 2 villages with a higher cancer incidence of every chosen town. Finally, there were 28 villages. 158 water samples from these villages were, respectively, collected in two water period: the high flow and low flow period, and three water sources: deep groundwater, shallow groundwater, and surface water. About 500 mL of water was bottled and HNOμg/L were defined as overproof in drinking water. We use SAS9.1 for data analysis. The data of incidence and mortality of population Cancer from 2008 to 2010 year come from Cancer Registration System of Huai'an city. The correlation analysis was used to study the relationship between the Mn concentration and cancer incidence and mortality.According to national standards for drinking water quality, Mn concentrations more than 100 5, 282.47/105, and 179.33/105 and 157.01/105, 192.20/105, and 170.02/105. There was a significant difference in cancer incidence and mortality among these three counties (P < 0.01). This study included 158 water samples from 28 villages of 14 towns in Chuzhou county, Jinhu county, and Xuyi county. All of them, 84 water samples were from the high flow period and 74 water samples from the low flow period. Detectable rate was 100%.Cancer incidence and mortality of Chuzhou county, Jinhu county, and Xuyi county was 216.54/10μg/L, the max was 4364.00 μg/L, and the mean was 452.32 μg/L ± 507.76 μg/L. The mean concentration of Mn in Chuzhou county, Jinhu county, and Xuyi county was, respectively, 401.15 μg/L ± 309.76 μg/L, 574.46 μg/L ± 701.60 μg/L, and 350.17 μg/L ± 352.26 μg/L. There was no significant difference in the mean concentration among these three counties . The mean concentration of Mn in the high flow period and the low flow period was 265.47 μg/L ± 388.83 μg/L, and 664.41 μg/L ± 544.82 μg/L, and there was a significant difference between them . The mean concentration of Mn in deep groundwater, shallow groundwater, and surface water was 287.83 μg/L ± 370.25 μg/L, 573.64 μg/L ± 672.47 μg/L, and 506.33 μg/L ± 362.47 μg/L, and there was a significant difference among them . The concentration of Mn in deep groundwater and shallow groundwater was more than in surface water; the difference was significant (P < 0.05). During the high flow period, the mean concentration of Mn in three water sources was difference significantly with the mean concentration in deep groundwater being only 72.01 μg/L ± 130.91 μg/L while the other two sources being 437.85 μg/L ± 439.79 μg/L, 286.56 μg/L ± 429.09 μg/L  (The min concentration of Mn in water samples was 0.02  < 0.05) .μg/L ± 439.79 μg/L, 286.56 μg/L ± 429.09 μg/L, and 72.01 μg/L ± 130.91 μg/L, and there was a significant difference among them . During high flow period, the mean concentration of Mn in deep groundwater, shallow groundwater, and surface water was 607.24 μg/L ± 168.38 μg/L, 871.35 μg/L ± 753.13 μg/L, and 503.65 μg/L ± 406.83 μg/L, and there was a significant difference among them . S-N-K showed that in high flow period Mn concentration in both shallow groundwater and surface water were higher than that in deep groundwater (P < 0.05). During high flow period, Mn concentration in shallow groundwater was higher than that in deep groundwater (P < 0.05)  .χ2 = 3.86, P > 0.05). The ratio of superscale of Mn in the high flow period and the low flow period was 53.57% and 100%; the difference was significant . The ratio of superscale in surface water was 87.23% which was the highest one. And next one was in shallow groundwater which was 80.00%. The third one was in deep groundwater which was 60.71%. The difference among them was significant   .χ2 = 19.24, P < 0.05)  .P < 0.05). Regression analysis showed that, for 1 μg/L increase in Mn concentration, there was a corresponding increase of 0.45/105 cancer cases and 0.35/105 cancer deaths (P < 0.05)  Tables .Our study showed that the mean concentration of Mn in drinking water in Huai'an city was much higher than the concentration; refer to standards for drinking water quality. The ratio of superscale is high of 75.32% and during the low flow period was 100%. We also found that the difference between it in the high flow period and low flow period was significant. For three water sources, the ratio of superscale of Mn in surface groundwater and shallow groundwater was higher than deep in water, while the same as the mean concentration. Excessive and prolonged exposure to Mn resulted from welding and mining, inhalation of combustion products from the antiknock agent in fuel (MMT) and highly concentrated Mn concentrations in ground/well water, and so on [μg/L increase in Mn concentration, there was a corresponding increase of 0.45/105 cancer cases and 0.35/105 cancer deaths. At first glance, this finding appears to be unbelievable. A large ecological study was carried out in North Carolina, and they also found manganese concentration in groundwater correlate with cancer mortality [Another important finding was that Mn concentration in drinking water in Huai'an city correlated with cancer incidence and mortality. In our city, for a 1 ortality . There aortality , 14. An ortality , 16.μg/d [Though World Health Organization (WHO) found among the general population, the average estimated Mn intake from drinking water is substantially lower than the intake of Mn from the diet, typically 20 μg/d , our finμg/d –21. ThusBecause data were analyzed at the population level and these same county-level factors may be related to individual adverse health behaviors such as cigarette smoking and obesity that contribute to cancer incidence and mortality, these ecological findings should be confirmed at the individual level or in animal models.The mean concentration of Mn in drinking water in Huai'an city was much higher than the concentration according to the national standards for drinking water quality. And the ratio of superscale is very high. Mn concentration is carcinogenic to people, which correlates with cancer incidence and mortality. Measures for mental pollution prevention and control must be taken immediately.


Cinnamomum cassia we performed randomized, double-blinded placebo-controlled study using euglycaemic-hyperinsulinaemic clamp.Published studies have reported conflicting results regarding the effects of cinnamon on glucose, lipids and insulin. To gain further insight into the metabolic effects of Twenty-one subjects with impaired glucose tolerance (IGT) were included in the study (10 or 11 subjects in each group). The study groups were matched for age, gender and body mass index (BMI). Waist-to-hip ratio, BMI, blood pressure, fasting blood glucose, insulin, triglycerides, total cholesterol, low-density lipoprotein, high-density lipoprotein , HbA1c, ASAT, ALAT, bilirubin, ALP, GT and PK were measured before and after the intake of capsules equivalent to 6 g cinnamon twice a day for 12 weeks. The changes in insulin resistance were measured by euglycaemic-hyperinsulinaemic clamp. The Wilcoxon signed rank sum test, the Mann-Whitney U test and Pearson’s chi-squared test were used to analyse the data. Values of p < 0.05 were considered to indicate statistically significant differences.At enrolment, the groups were similar in terms of age, gender and BMI. Of the 21 randomized patients with IGT, 17 completed the study (8 controls vs. 9 treated). The ingestion of 6 g cinnamon twice a day for 12 weeks had no significant effect on insulin sensitivity, HbA1c, fasting glucose or BMI. No significant changes were seen in lipids or liver enzymes.C. cassia twice a day for 12 weeks did not change the insulin sensitivity or liver enzymes in subjects with IGT.This study showed that ingestion of 6 g  Chronic diseases such as cardiovascular disease, cancer, chronic respiratory disease and diabetes mellitus are currently the major cause of death in almost every country worldwide, and not only in the developed countries. According to the World Health Organization chronic diseases cause almost 35 million deaths each year worldwide, which is 60% of the total mortality. Cardiovascular diseases account for almost 50% of deaths due to chronic diseases . The agiImpaired glucose tolerance (IGT) is the term used to describe the intermediate stage between normal glucose metabolism and type 2 diabetes mellitus. Subjects with IGT have an increased risk of developing type 2 diabetes mellitus and cardiovascular disease compared with normoglycaemic subjects, and early interventions are, therefore, important. The causes of type 2 diabetes mellitus are multifactorial, but diet plays an important role in the incidence, severity and management of the condition . The relCinnamomum cassia. C. cassia is obtained from the bark of the outer skin of a tall evergreen tree, and contains active components including cinnamic aldehyde, cinnamyl aldehyde, tannin, mucus and carbohydrates. These components have been found to have anti-oxidant, anti-microbial, anti-inflammation, anti-diabetic and anti-tumour activity [C. cassia and C. zeylanicum extracts have been shown to be among the most effective natural substances in the regulation of blood glucose [C. cassia, may have detrimental effects on the liver, and the Federal Institute for Risk Assessment in Europe has, therefore, suggested that it be replaced by C. zeylanicum[C. cassia powder reduces the postprandial blood glucose concentration [C. zeylanicum did not affect postprandial blood glucose or insulin levels in humans [Various types of natural remedies have been used historically for the treatment of ailments, among them activity –8. C. ca glucose . Howeverylanicum. We haventration . Howevern humans .C. cassia increased insulin sensitivity in subjects with IGT, using the euglycemic-hyperinsulinaemic clamp, which provides a direct measure of insulin resistance. To the best of our knowledge, the direct measurement of the effect of cinnamon on insulin resistance been not been studied previously in humans. We also investigated whether C. Cassia supplementation had any effect on liver enzymes.Other studies have shown conflicting results regarding the effect of cinnamon on insulin sensitivity, when measured indirectly –16. In tTwenty-one subjects with IGT, who met the study criteria, were included in this randomized, double-blinded placebo-controlled study. The inclusion criterion was diagnosis of IGT, by a standard 75 g OGTT, for less than 12 months before enrolment. Glucose tolerance status and fasting blood glucose levels were evaluated using the criteria established by the WHO [C. cassia and 200 mg cellulose. Participants received both individual verbal and written information and signed a consent form. The participants were instructed to ingest one capsule each morning at breakfast time and one each evening at dinner time for 12 weeks. During this period, the subjects were examined three times, at baseline, after sex weeks and finally, after 12 weeks. Insulin sensitivity was measured at baseline and after 12 weeks, in the morning after a 12-hour fast. Smoking and snuff-taking were prohibited 8 hours prior to and during the test.All the capsules were prepared in advance by Scandinavian Nutrients AB , and contained either 700 mg cellulose or 500 mg 2 body surface area/min. After 4 min, glucose infusion (200 mg/ml) was started; the infusion rate being adjusted manually throughout the procedure to maintain the blood glucose level at 5.0 mmol/l. Blood glucose was determined at the bedside every 5 minutes. Blood samples were taken after 60 and 120 min for the analysis of the insulin concentration. Blood glucose concentrations were measured with the HemoCue Glucose system . The precision of the HemoCue Glucose system was better than 0.3 SD in the range 0 mmol/l to 22.2 mmol/l.Insulin sensitivity was determined with the euglycaemic-hyperinsulinaemic clamp, according to DeFronzo et al. . IntraveAll analyses of plasma and whole blood were performed on samples obtained after overnight fasting. Analyses of fasting plasma triglycerides, total cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL), HbA1c, ASAT, ALAT, bilirubin, ALP, GT, PK, whole-blood glucose (fasting blood glucose), and insulin were carried out at the time of baseline examination, at the Department of Clinical Chemistry, Skane University Hospital in Malmö, which is affiliated to a national standardization and quality control system. Insulin concentrations were measured using an immunoassay with an alkaline phosphatase conjugate . The sensitivity of the insulin immunoassay was 0.03 mU/l, and the intra-assay coefficient of variation was less than 10% in the range 0.03 mU/l to 300 mU/l. Total cholesterol and HDL were measured by enzyme assay kits, using an automated analyser . LDL was calculated using the Friedewald equation . All samThe study was approved by the Ethics Committee of Lund University and was performed according to the Helsinki Declaration. The study started in 2011 and was completed in 2013.All statistical calculations were performed using SPSS for Windows software . The Wilcoxon signed rank sum test was used to compare quantitative variables within the group, and the Mann-Whitney U test was used to compare quantitative variables between groups. Pearson’s chi-squared test for categorized variables was used to test for statistical significances between the groups. Values of p < 0.05 were considered to indicate statistically significant differences.No significant changes were seen in clinical or demographic characteristics such as BMI, waist:hip ratio, systolic or diastolic blood pressure between the two groups at baseline Table . No signC. cassia supplementation (12 g/d) did not improve fasting plasma glucose, insulin, blood lipids or insulin sensitivity in patients with IGT. Neither were any of the liver enzymes affected in this study. Although the effects of cinnamon on glucose, insulin resistance and lipid profile have been investigated in several studies, showing conflicting results, to the best of our knowledge, this is the first study to use a direct measurement of insulin resistance.In this study, it was found that 12 weeks of in vitro and in vivo[, and cinnamon may have qualities that decrease insulin resistance. However, this has not been verified in humans using direct measurements, i.e. the euglycaemic-hyperinsulinaemic clamp. A reduction in insulin resistance was seen in six nondiabetic women after the intake of C. burmannii, 1 g per day, in capsules, for 8 weeks [C. cassia  [C. cassia per day for 6 weeks [Cinnamon has demonstrated qualities that enhance glucose uptake by activating insulin receptor kinase activity, autophosphorylation of the insulin receptor, and glycogen synthase activity,  in vivo–22. In a in vivo. This ma 8 weeks . A reduc 8 weeks . Solomonper day) . However 6 weeks . In the  6 weeks –26. Howe 6 weeks  or who h 6 weeks . On the  6 weeks . Methods 6 weeks , 31. AltC. cassia powder per day for 40 days. Not only were the levels of fasting glucose reduced in subjects with type 2 diabetes mellitus, but positive effects were also seen on triglyceride, LDL and total cholesterol levels [C. cassia supplement per day for 4 weeks reduced fasting glucose in type 2 diabetes mellitus patients. In contrast, Vanschoonbeek et al. and Blevins et al. found no difference in fasting glucose in type 2 diabetes mellitus patients after taking 1.5 g C. cassia per day for 6 weeks, and 1 g C. cassia per day for 12 weeks, respectively [In 2003, Khan et al. reported remarkable results following the ingestion of 1, 3 and 6 g l levels . Followil levels  revealedectively , 33. No ectively , 32, 33.ectively .C. cassia per day for 12 weeks ) [C. cassia per day for 12 weeks) and Lu (120 mg per day or 360 mg C. aromaticum per day for 12 weeks) [However, in a recent study, HbA1c was found to be decreased in patients with type 2 diabetes mellitus (1 g  weeks ) . Similar2 weeks) . In thes2 weeks) . These cC. cassia. Debate on the toxicity of coumarins started after experimental studies on hepatotoxicity in dogs. The Federal Institute for Risk Assessment issued a warning on the consumption of cinnamon and established a limit of 0.1 mg coumarin per kg body weight/day. In a study on 114 participants who ingested 30 mg coumarin combined with a vasoactive component, Schmeck-Lindenau et al. found that nine of the patients showed elevated levels of transaminases in serum [C. cassia[C. cassia is 3000 mg/kg. In our study, where the subjects ingested 12 g C. cassia per day for 12 weeks, i.e., about 36 mg coumarin, no significant changes were seen in transaminases. Askari et al. found a reduction in the level of transaminases in patients with nonalcoholic fatty liver disease, who were given 1.5 g cinnamon/day for 12 weeks [Coumarin is present in cinnamon, especially in serum . But the. cassia. AccordiThe strengths of the present study were the double-blind placebo-controlled design with few drop-outs, and the direct measurement of insulin sensitivity. However, this study also had some limitations. Firstly, the sample size was small and all the participants were living in southern Sweden. Secondly, the control group had a significantly lower insulin response than the treated group after 12 weeks, which indicates that the placebo capsules had an active effect. The placebo capsules contained cellulose, which is a type of fibre, and the amount of fibre was probably sufficient to cause an effect on the gastrointestinal tract, leading to an indirect effect on insulin response. The relation between high fibre diets and a low insulin response is already known . ThirdlyC. cassia twice a day for 12 weeks did not change the insulin sensitivity or liver enzymes in subjects with IGT.In conclusion, the findings of this study were that the ingestion of 6 g


Cystatin C, serum creatinine (SCr) levels, and estimated glomerular filtration rates (eGFR) were assessed at several time points. The primary endpoint was CIN incidence; secondary endpoint included changes in cystatin C, SCr, and eGFR. CIN incidence was significantly lower in the BNP group compared to controls . In addition, a more significant deterioration of eGFR, cystatin C, and SCr from 48 h to 1 week (P < 0.05) was observed in controls compared to the BNP group. Although eGFR gradually deteriorated in both groups, a faster recovery was achieved in the BNP group. Multivariate logistic regression revealed that using >100 mL of CM  and BNP administration  were independently associated with CIN. Combined with hydration, exogenous BNP administration before CM effectively decreases CIN incidence in CKD patients.The role of brain natriuretic peptide (BNP) in the prevention of contrast-induced nephropathy (CIN) is unknown. This study aimed to investigate BNP's effect on CIN in chronic kidney disease (CKD) patients undergoing elective percutaneous coronary intervention (PCI) or coronary angiography (CAG). The patients were randomized to BNP (0.005  The modified intention-to-treat population was analyzed. All statistical analyses were performed using SPSS 13.0 . Data are presented as mean ± standard deviation (SD) or median (interquartile ranges) for continuous variables and proportions for categorical variables. The independent samples t-test was used to compare continuous variables and the χ2-test or Fisher's exact test for categorical variables. Repeated measure ANOVA and the Bonferroni post hoc test were used to evaluate the changes in variables within the same group. Multivariate logistic regression analysis was used to identify independent predictors of CIN. Two-sided P values <0.05 were considered statistically significant.Sample size , 209 completed the study. Before starting the trial, one patient suffered from hypotension; two patients had heart dysfunction ; one patient suffered from refractory angina and needed early invasive therapy before the study; one patient had end-stage renal failure; and four patients were lost to follow-up after the procedure. Finally, 106 and 103 patients were analyzed in the BNP and control groups, respectively . The incidence rates of a SCr increase of ≥0.5 mg/dL were 3.8% and 13.6% in BNP-treated patients and controls, respectively (P = 0.011). When defined as a SCr increase of ≥25%, the incidence of CIN was also lower in the BNP group compared to the control group   .P = 0.251), SCr , and eGFR  were similar between the two groups. Nevertheless, a more significant deterioration was observed in the control group compared to BNP-treated patients. Cystatin C levels for the control and BNP groups were 1.56 ± 0.29 mg/L and 1.75 ± 0.94 mg/L  and 1.20 ± 0.24 mg/L and 1.88 ± 0.82 mg/L , respectively. Meanwhile, 140.1 ± 13.9 μmol/L and 151.2 ± 15.9 μmol/L were observed for SCr in the control and BNP groups at 48 h (P = 0.017), respectively, and 123.8 ± 14.4 μmol/L and 159.7 ± 13.8 μmol/L, respectively, at 1 week (P < 0.001). eGFR of 43.6 ± 17.1 mL/min and 40.2 ± 18.7 mL/min were observed in the control and BNP groups, respectively, at 48 h (P = 0.046) and 50.4 ± 14.9 mL/min and 37.9 ± 15.9 mL/min at 1 week (P < 0.001). There were no significant differences between the two groups for the mean eGFR, cystatin C, and SCr values at 1 month.As shown in Overall, eGFR gradually deteriorated until 1 week, followed by a mild improvement at 1 month in the control group, whereas the deterioration observed from 24 h to 48 h in the BNP group was completely restored at 1 week, and values remained at baseline until 1 month .Besides the nine patients who suffered from complications before starting the trial, no patient presented adverse effects related to the use of rhBNP.P = 0.023), to have a history of diabetes , to have hypertension , to have received PCI (P = 0.014), to have received more CM , and to be non-BNP-treated (P < 0.001). After multivariate logistic regression analysis, only the CM volume (CM >100 mL) and BNP administration were significant independent predictors of CIN  or absolute (≥0.5 mg/dL) increase in SCr levels from baseline. Basically, SCr rises within the first 24–48 hours after CM exposure, peaks at 3–5 days, and returns near baseline values within 1–3 weeks . However5% or absApart from BNP administration, CM volume >100 mL was found to be an independent predictor of CIN in this study. This finding is supported by previous reports demonstrating that large CM volumes are associated with an increased risk of CIN , 22, 23.A few limitations of the present study should be mentioned. First, the relatively small sample size prevents the generalization of our findings. In addition, this was a single center trial with the inherent selection bias. Finally, the study was not blinded. Therefore, multicenter, randomized, controlled trials with a larger sample size are required to further evaluate the beneficial effects of BNP on CIN.Overall, this study showed that, in addition to hydration, exogenous administration of BNP before CM exposure is effective in decreasing the incidence of CIN in patients with CKD.


Peripheral cytopenias are common in systemic lupus erythematosus (SLE), but bone marrow involvement is rarely reported. Myelofibrosis is a rare disorder characterized by reticulin fibrosis of the bone marrow, which usually occurs in response to clonal proliferation of hematopoietic stem cells in myeloproliferative disorders. However, bone marrow fibrosis has also been described in association with auto-immune diseases, especially SLE.We will report here a new case of bone marrow fibrosis associated with SLE. We also reviewed the 27 cases published in the English language literature, and will discuss the clinical presentation, outcome, treatment, and pathophysiology of bone marrow fibrosis occurring in association with SLE.Over one half of patients were diagnosed concomitantly with bone marrow fibrosis and SLE. Epidemiological, clinical and biological features of lupus were unremarkable. Except for the presence of reticulin fibrosis, the findings from the bone marrow biopsies proved highly variable. Overall mortality was about 14% but corticosteroid-based therapy lead to clinical improvement and reverted bone marrow fibrosis in most cases. Data on the usefulness of other immunomodulatory therapies are inconclusive.SLE may be complicated by bone marrow involvement, of a likely autoimmune origin. Bone marrow fibrosis occurring with SLE is probably similar to “primary autoimmune myelofibrosis” and may respond to steroid and immunomodulatory therapies. Further studies with standardised proofreading of bone marrow aspirations and biopsies are needed to delineate the clinical and biological features of this rare complication of SLE. Hematological abnormalities such as anemia, auto-immune hemolysis, leukopenia, lymphopenia and thrombocytopenia are very common in systemic lupus erythematosus (SLE)  without anemia or thrombocytopenia. Antinuclear antibodies were present (>1/1280 with a speckled fluorescence pattern). Anti-DNA antibodies were absent. Anti-U1-RNP and anti-SSA antibodies were positive. Three years later (2002), she was admitted to the hospital because of atypical eating disorder and psychotic behaviour. No evidence for neuropsychiatric lupus was found despite an extensive work-up. The patient was diagnosed with schizophrenia and treated with olanzapine. Nine years later (2011), she was referred to the hospital for fatigue, fever and pancytopenia. With the exception of extreme paleness, clinical examination was unremarkable. Her weight is normal. Laboratory findings on admission showed the following values: leukocyte count 0,48 × 109/l with 0.28 × 109/l neutrophil granulocytes and 0.08 × 109/l lymphocytes, hemoglobin 2.9 g/dl, platelet count 15 × 109/l and reticulocyte count 3 × 109/l, low-normal level of folate, normal levels of vitamin B12 and ferritin. Anti-SSA, anti-RNP 70 and anti-U1-RNP antibodies were positive. The anti-DNA antibody titer was 76 UI/ml in ELISA [normal range < 10]. Direct Coombs’ test was negative. C3, C4, and CH50 were respectively 1.18 g/l [normal range: 0.79–1.52], 0.12 g/l [normal range: 0.2–0.51] and 120%. Abdominal ultrasonography showed limit homogen splenomegaly (around 12 cm). Tear drop cells or leukoerythroblastic blood smear were not noted. Two attempts to aspirate bone marrow at different sites were unsuccessful and yielded only dry taps. Bone marrow biopsy showed hypercellular marrow (cellularity: 80%) with focal lymphocytic infiltration, dysmyelopoiesis, erythrophagocytosis and grade 1–2 fibrosis. JAK-2, MPL W515L/K, and calreticulin mutations were not screen. A diagnosis of SLE with bone marrow involvement was retained. High dose methylprednisolone (500 mg/d) was given for 3 days. Then hydroxychloroquine and prednisone (1 mg/kg) were started, in addition to supportive care with blood transfusion and antibiotics. However, the patient remained pancytopenic. Intravenous immune globulins (30 g/d) were given for 4 days with remarkable improvement. Hemoglobin level after 2 months was 11.8 g/dl, platelet count was 175 × 109/l and leucocyte count was 3.27 × 109/l with 2.32 × 109/l neutrophil granulocytes and 0.49 × 109/l lymphocytes. Prednisone was continued for 17 months and gradually tapered. The patient did not experience change in her mental status during the course of steroid treatment. She remains in good physical health and still takes hydroxychloroquine and olanzapine.A 17 year-old woman with a history of polyarthralgias and photosentivity was referred to the hospital in 1999 for the evaluation of a Raynaud’s phenomenon. Clinical examination revealed no anomaly. She presented with neutropenia , 13/28 had bicytopenia (anemia or/and leukopenia or/and thrombocytopenia), 1/28 had thrombocytopenia and neutropenia without leukopenia and 1/28, only thrombocytopenia. Neutropenia  was observed in 10/12 (16 missing data) patients, lymphopenia in 4/9 (19 missing data), hemolysis with hyper-reticulocytosis in one case and a positive direct Coombs’ test in 10/18 (10 missing data). Hemoglobin levels ranged from 13.1 to 2.7 g/dl (mean 7.3 g/dl), platelets from 341 to 1 × 109/l (mean 50 × 109/l), and leucocytes from 6.8 to 0.35 × 109/l (mean 3.4 × 109/l). Of the 24 patients with thrombocytopenia, 11 had deep (<20 × 109/l), 8 severe (<50 × 109/l), and 5 moderate thrombocytopenia. Tear drop cells or leukoerythroblastic blood smear, two common finding in primary myelofibrosis, were noted in 14 patients.Thirteen of the 28 patients had pancytopenia  to normal (11/28) or decreased (5/28) without any increase in blasts. All elements including megakaryocytes appeared morphologically normal. Megakaryocytes were increased or/and with clustering in 13/28 patients and decreased in 1/28. Focal or massive lymphocytic infiltration was observed in 4/28, plasmocytosis in 1/28 and erythroid hyperplasia in 2/28. Fifteen patients underwent repeated bone marrow examination showing improvement, with reduction in reticulin amounts in 12/15.Antinuclear antibodies were found in all patients, anti-dsDNA in 12/28, anti-Ro/SSA in 2/28, anti-histone in 1/28, and a low complement level in 18/28 patients. Antiphospholipid antibodies were detected in 4/28 patients. Anti-platelet antibodies testing came out positive in 3 patients and negative in 4.Follow-up time ranged from a few months to years but was often unspecified. The overall mortality was 14% (4/28). Two patients died within few days with no other treatment than blood transfusions and antibiotics. Two other patients who died received only prednisone as a specific treatment. Improvement was noted in 17/28 patients, transient response with need for new treatment in 5/28, and no improvement in 2/28. Supportive care with antibiotics and transfusions was explicitly mentioned for 8 patients. Two patients received granulocyte colony stimulating factor (G-CSF).Immunomodulatory therapies consisted in corticosteroids (26/28) , intravenous immune globulins (4/28)  and is more common in males (ratio 3:2) (Tefferi et al.Although it is likely that negative outcomes are less frequently reported in case reports, for which follow up data may be lacking, and that consequently the overall mortality may be higher than the 14% documented from this review, this mortality rate suggests a more favourable course for SLE-associated bone marrow fibrosis than for primary myelofibrosis (Tefferi et al.SLE may be complicated by bone marrow fibrosis, which is likely to be of autoimmune origin. This feature may be more common than previously thought, with cases being incorrectly characterized as blood peripheral cytopenias in patients previously diagnosed with SLE, and cases being misdiagnosed with primary myelofibrosis in patients not previously diagnosed with SLE. We think that in patients with SLE, cytopenias should be confirmed by bone marrow aspiration, and by bone marrow biopsy in atypical or refractory cases. Moreover, autoimmune myelofibrosis or SLE-associated bone marrow fibrosis should be considered in cases of primary myelofibrosis with atypical features such as young age and female sex, absence of spleen enlargement, or absence of JAK2 V617F mutation, because this condition seems amenable to efficient treatment. High-dose corticosteroid therapy with or without intravenous immune globulins should be the first-line therapy.In order to improve knowledge of bone marrow involvement in SLE, we have established a French registry, with centralized proofreading of bone marrow aspirations and biopsies. We hope to achieve a sufficient sample size for epidemiological and clinical research on this unusual feature of lupus.Written informed consent was obtained from the patient for the publication of this report.


Aloe vera (AV) using probiotics. To explore the potential benefits, the antioxidant effects and the antibacterial effects on foodborne pathogens of Aloe fermentation supernatant were evaluated in vitro. Our results indicated that the Aloe fermentation supernatant fermented by Lactobacillus plantarum HM218749.1 had very strong scavenging capacities of the DPPH (86%), O2•− (85%), •OH (76%), and Fe2+ chelation (82%) and reducing powers (242.5 mg/L), and the inhibition zones for Salmonella typhimurium, Salmonella enteritidis, Shigella flexneri, Escherichia coli, Listeria monocytogenes, S. dysenteriae 301, Staphylococcus aureus Cowan1, and Propionibacterium acnes were 16, 15, 19, 20, 21, 20, and 27 mm. Moreover, the low concentration of Aloe fermentation supernatant had significantly reduced the production of IL-1β, TNF-α, and IL-6 in both mRNA and protein levels (P < 0.01). Therefore, the Aloe fermentation supernatant can be used as functional beverage or cosmetic ingredients to guard human intestinal health, delaying senescence, and prevent chronic diseases.Little work is done to develop Aloe vera (AV) is a cactus-like plant which has been used widely in herbal medicines for millennia [illennia . The comillennia –6. Now, illennia . HoweverProbiotics are defined as “live microorganisms which when administered in adequate amounts, confer a health benefit on the host.” Previous studies indicated that probiotics conferred miscellaneous health benefits include an improvement in lactose intolerance, an increase in natural resistance to infectious disease in the gastrointestinal tract, the suppression of cancer, an antidiabetic agent, and a decrease in serum cholesterol levels –9. In re Lactobacillus plantarum is pointed out as an industrially important microorganism that can be found and isolated from dairy products, fermented foods , environments , human mouth, intestinal tract, and stools [ L. plantarum HM218749.1 was isolated from human intestinal, which could grow better at 37°C in an anaerobic environment and could resist harsh conditions of gastrointestinal tract . Moreover, this strain could strongly adhere to HT-29 cells and reduce the adhesion of Shigella dysenteriae 2457, Staphylococcus aureus Cowan1, Enterobacter sakazakii 45401, and Escherichia coli 44102 to HT-29 cells, and its long-term residence in mice intestine increased the number of lactobacilli and decreased the number of enterococci [Among the probiotics,d stools , 14, andd stools . In our d stools , L. planerococci . Bifidobacterium longum ATCC15708, L. plantarum HM218749, L. rhamnosus GG, and Streptococcus thermophilus G1) grew well in AV substrate and conferred AV sound antioxidant and antibacterial effects, and the L. plantarum HM218749.1 was finally chosen for its popularity in food fields and perfect beneficial effects verified in our previous studies, and the cell experiment was carried out to further verify the anti-inflammatory effects of the Aloe fermentation supernatant.In the present study, the tested probiotics , L. plantarum HM218749 (isolated from human intestine), L. rhamnosus GG (isolated from the milk), and Streptococcus thermophilus G1 (isolated from the milk) were cultured in de Man Rogosa Sharpe (MRS) medium (for Lactobacilli and Bifidobacterium) and brain/heart infusion (BHI) medium (for Streptococcus thermophilus G1) for 24 h, and 108 cfu/mL of the strains was used as an inoculum to be added to Aloe vera, Aloe vera + 5% glucose, or Aloe vera + 5% glucose + 5% skimmed milk for preparing the Aloe fermentation supernatant, which was incubated for 15 to 18 h at 37°C or 42°C. Finally, the Aloe fermentation supernatant and AV were freeze-dried using vacuum freeze drier under with the following conditions: freeze drying at 0.2 mbar vacuum, −40°C for 48 h. The lyophilized powder was stored at −20°C for further use.The probiotic2, 5% H2, and 10% CO2) at 37°C or 42°C for 24~36 h followed by counting colonies from plates showing 25–250 colonies [Bacterial counts were enumerated as previously reported . Dilutedcolonies . Aloe fermentation supernatant, 10% of the probiotics were added to AV extract and fermented for 15 to 18 h at 37°C, and then the Aloe fermentation cultures were centrifuged at 4000 ×g for 10 min, and the pellets in the bottom of the centrifuge tubes  were deemed as the residual substance, and the clear water insoluble substance was deemed as supernatant, and the AV extract was used as a control. Then, the clearance of the DPPH, O2•−, •OH, Fe2+ chelation, and oxygen-reduction activity were measured just as in [ Salmonella typhimurium ATCC 13311, S. enteritidis ATCC13076, S. flexneri ATCC 12022, E. coli 44102, Listeria monocytogenes ATCC 19111, S. dysenteriae 301, S. aureus Cowan1, and Propionibacterium acnes ATCC 11827 were spread on the surface of the LB agar plates, and the culture supernatant (200 μL) was loaded into an Oxford cup , which was placed on the surface of the agar. The size of the inhibition zone was measured until a formation of clear zone around the Oxford cup. The experiment was carried out in duplicate [For preparation of theuplicate .μg/mL of streptomycin) at 37°C in a 5% CO2 incubator. Emodin , polysaccharide , Aloe vera (lyophilized powder), and Aloe vera + L. plantarum (lyophilized powder) were dissolved in phosphate-buffered saline (PBS) solution and their effect on cell viability of RAW 264.7 cells was determined by MTT assay [RAW 264.7 cells (passages 8–20) were maintained in DMEM supplemented with 10% FBS and antibiotics , polysaccharide , Aloe vera , and Aloe vera + L. plantarum  in 96-well microtiter plates for 36 h at 37°C in a humidified incubator. Control cells treated with PBS served as the vehicle group. Each concentration of drugs was repeated in 5 wells. After incubation for specified times, MTT reagent  was added to each well and incubated for 4 h. After careful removal of the MTT solution, 150 μL of DMSO was added to each well. The absorbance was recorded on a microplate reader at the wavelength of 570 nm.RAW 264.7 cells were plated at a density of 10000 per well in 100 4 cells per well) for 24 h and were then washed with PBS. Cells were treated with/without lipopolysaccharide , emodin (2.5 μg/mL and 10 μg/mL), polysaccharide (2.5 μg/mL and 15 μg/mL), Aloe vera (5 μg/mL and 50 μg/mL), and Aloe vera + L. plantarum (5 μg/mL and 50 μg/mL) for 24 h, and then the cells and supernatants were harvested for Q-PCR and ELISA analysis, respectively.RAW 264.7 cells were seeded into 24-well plates . Quantitative real-time PCR was performed with 7900HT fast real-time PCR system  using 2 × SYBR Green master mix (Bio-Rad). Forty cycles were conducted as follows: 95°C for 30 s and 60°C for 30 s, preceded by 1 min at 95°C for polymerase activation with the following primers .For the evaluation of cytokine mRNA expression levels, total RNAs from the RAW 264.7 cells were prepared by adding TRIzol reagent  according to the manufacturer's protocol, and 2.4 β, TNF-α, and IL-6 in cell supernatants were determined using the ELISA kit for IL-1β (eBioscience), TNF-α (eBioscience), and IL-6 (eBioscience).The products of IL-1P < 0.05 or 0.01).Data are reported as means ± SD, and results were analyzed using SPSS 13.0 software  by means of independent one-way ANOVA tests in each sampling point. The differences among the three groups were assessed by means of the least significant difference (LSD) multiple comparison test ( B. longum (MRS medium), L. plantarum (MRS medium), L. rhamnosus LGG (MRS medium), and S. thermophilus (BHI medium) were inoculated in the corresponding media and cultured at 25°C, 30°C, 37°C, and 42°C, and the results showed that the optimum temperature for B. longum, L. plantarum, and L. rhamnosus LGG was 37°C, and S. thermophilus received its maximum yield at 42°C , and the growth promoting effect of the 5% glucose + 5% skimmed milk was much better than 5% glucose (b < 0.01)  .To evaluate the additive of the probiotics on the perfect antioxidant and antibacterial activities of AV , 20–22, 2•− clearance, Fe2+ chelation, and reduction activity, while the L. plantarum HM218749.1 had significantly (P < 0.01) enhanced the DPPH clearance and •OH clearance , while the addition of 5% glucose conferred the Aloe fermentation supernatant a perfect inhibition to all the tested pathogens, for example, S. typhimurium ATCC 13311 (inhibition zone diameter: 16 mm), S. enteritidis ATCC13076 (IZD: 15 mm), S. flexneri ATCC 12022 (IZD: 19 mm), E. coli 44102 (IZD: 11 mm), L. monocytogenes ATCC 19111 (IZD: 20 mm), S. dysenteriae 301 (IZD: 21 mm), and S. aureus COWAN1 (IZD: 20 mm) , which leads to an accumulation of the skins natural oils, bacteria, and acne vulgaris [Interestingly, no antimicrobial effect was observed using AV extract and: 20 mm) . Most ofvulgaris .β, IL-6, and TNF-α) lead to the damaging of cells and tissues and the activation of macrophages in inflammation-associated diseases  [During inflammation, excess levels of cytokines (IL-1patitis) , 25. Aloe fermentation supernatant on the production of IL-1β, IL-6, and TNF-α, different concentrations of emodin , polysaccharide , AV extract , and Aloe fermentation supernatant  were cocultured with RAW 264.7 cells to test their toxicity on RAW 264.7 cells, and the results indicated that no obvious toxicity was observed for all the tested materials, and the low concentration of emodin (1 μg/mL) showed promoting effects on the growth of RAW 264.7 cells , O2•− (85%), •OH (76%), Fe2+ chelation (82%), and reducing powers (242.5 mg/L) . Therefore, the sound inhibition of the growth of P. acnes ATCC 11827 (causative bacteria for acne vulgaris) and antioxidant activity made the Aloe fermentation supernatant a potential ingredient for cosmetic.After a series of evaluations, the.5 mg/L) . Moreoveβ, IL-6, and TNF-α induced by activated inflammatory cells  will lead to the damage of cells and tissues, which eventually cause the inflammation-associated diseases; for example, rheumatoid arthritis, chronic hepatitis, and macrophages play an important role in regulating several immunopathological conditions and inducing overexpression of the proinflammatory mediators of the inflammatory process [ Aloe fermentation supernatant fermented by L. plantarum HM218749.1.As we know, excess levels of IL-1 process , 25. So  Aloe fermentation supernatant in mRNA level and protein level . In addition, the L. plantarum HM218749.1 could remove the bitter taste of AV, and our human results (n = 15) indicated that the Aloe fermentation supernatant had shortened the acne cure time and had a perfect whitening effect (data unshown).The results indicated that both the emodin (anticancer activity)  and the  Figures  and 5. It effect , its antal tract  and endo Aloe fermentation supernatant's sound antimicrobial effect and anti-inflammatory effect, together with its intestinal health promoting effect when L. plantarum HM218749.1 enters host intestines, indicate its potential use as functional foods or cosmetics to guard human intestinal health, delaying senescence and preventing chronic diseases.In conclusion, the