Objective:To evaluate the effect of epigallocatechin-3-gallate (EGCG) and epigallocatechin-3-O-(3-O-methyl)-gallate (EGCG-3Me) on the anti-bacterial effect and the stability of intraradicular dentin-adhesive interface.Methods:EGCG and EGCG3Me with the concentration of 400 μg/ml were incorporated into Single Bond 2 (SB2) respectively to obtain 2 modified adhesives E-SB2 and E3-SB2.Confocal laser scanning microscopy(CLSM) and ultraviolet spectrophotometry were used to evaluate the anti-bacterial effect of the modified adhesives.Micro-Raman spectrum was used to test the degree of conversion (DC) of the adhesives.Push-out bond strength test was conducted to examine the immediate bond strength and the bond strength after themocycling.Results:E-SB2 and E3-SB2 both showed inhibiting effect on the proliferation of E.faecalis,while E3-SB2 performed stronger inhibiting effect.DC and the immediate push-out bond strength of SB2 were not decreased with the incorporation of EGCG or EGCG-3Me(P ＞ 0.05).E-SB2 and E3-SB2 showed significantly higher push-out bond strengths than that of SB2 (P ＜ 0.05) after themocycling.Conclusion:EGCG and EGCG-3Me modified adhesives have anti-bacterial effect and can enhance the stability of bonding between intraradicular dentin and adhesive,EGCG-3Me may have stronger anti-bacterial effect.

A technique of diffusive gradients in thin films (DGT) was developed for the in situ measurement of reactive phosphorus species in natural waters, sediments and potentially soils. Polyacrylamide / basic magnesium carbonate was used as the novel binding phase of DGT. Various factors, such as initial concentration, deployment time, pH and ionic strength, which may affect the adsorption of phosphate to the DGT were investigated. H2 SO4(0. 25 mol/ L, 10 mL) was used for elution of phosphate from the binding gel, and an elution efficiency of 85±5% was obtained. The DGT measurement was independent of ionic strength (0. 001-0. 05 mol/ L) and pH (4. 10-9. 15). The results indicated that the maximum adsorption capacities of DGT were limited to 20. 4 μg per disc ( T = 25℃, pH = 7. 00, [ P] = 2 mg / L). Good agreement was obtained between the measurement results of DGT method and molybdenum blue method in the P concentration from 0. 001 to 20 mg / L. The method detection limit (MDL) was 102. 4 ng / L. Field performances of DGT in synthetic seawater, the coastal seawater of Xiamen, Lake Yihai, Lake Chaohu and Nanfei River indicated that the basic magnesium carbonate-DGT method was more reliable than the commonly used ferrihydrite-DGT method.

Objective:To summarize the clinical and imaging features of five patients of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) with cysteine-sparing NOTCH3 gene missense mutations and explore potential pathogenicity of gene mutations.Methods:The clinical data from five patients who were admitted to the People′s Hospital of Zhengzhou University from March 2017 to November 2018 were collected. The patients were found to carry cysteine-sparing NOTCH3 gene mutations through genetic testing and diagnosed pathologically. They were probands confirmed from five unrelated family and all five patients were performed full exon detection and skin biopsy.Results:Genetic testing identified five patients with cysteine-sparing NOTCH3 gene missense mutations, a total of five different mutations, including p.R75Q, p.D80G, p.V237M, p.S1418L and p.R1761H. The first three mutations were found in the epidermal growth factor-like repeats (EGFr), the latter two mutations near the transmembrane domain. Granular osmiophilic material was identified in all cases examined with skin biopsy. The age at initial symptom onset of these five cases was ranged from 22 to 58 years and three cases presented cardiovascular risk factors. The primary clinical manifestations included migraine in one case, ischemic stroke in three cases, psychiatric disturbances in four cases, cognitive dysfunction in five cases, while gait disturbance, pseudobulbar palsy, and seizures accounted for only one case each. Magnetic resonance imaging of five patients all showed white matter hyperintensities (WMLs) and lacunar infarcts, and WMLs involved the anterior temporal pole and external capsules in three cases separately. According to the criteria proposed by Mui?o et al for evaluating the pathogenicity of cysteine-sparing NOTCH3 mutations, all five mutations are potentially pathogenic.Conclusions:Most characteristics of CADASIL patients with cysteine-sparing NOTCH3 gene mutations are similar to those of CADASIL patients with cysteine NOTCH3 gene mutations. Mutations not involving the EGFr may also have potential pathogenicity, and the specific mechanism still needs further study.

Objective:To explore the safety, efficacy and preliminary clinical application of the single plantar approach or in combination with the dorsalis pedis approach in the treatment of Lisfranc injury with poor dorsalis pedis soft tissue, metatarsal avulsion fracture or complicated multi-column lesions.Methods:(1) Six fresh cadaveric specimens of adult foot were collected and dissected through the plantar approach in order to determine the skin incision of the plantar approach and the safe area for plate-screw internal fixation, including start-stop points and courses of plantar nerves, blood vessels, tendons and ligaments, followed by plate-screw fixation on the specimens. (2) After feasibility of the plantar approach was confirmed by our anatomical study, it was used to treat the 3 patients who were admitted to Department of Orthopedics, The Third Hospital Affiliated to Southern Medical University between September 2020 and November 2021 for Lisfranc injury with severe necrosis due to dorsalis pedis skin contusion or metatarsal base avulsion fracture. They were 2 males and one female, with an average age of 51 years (from 34 to 68 years). The preliminary clinical efficacy was evaluated in terms of visual analogue scale (VAS), midfoot score of American Orthopaedic Foot and Ankle Surgeons (AOFAS), Maryland score, Kofoed score, fracture healing at the last follow-up and postoperative complications.Results:(1) Regarding the anatomical exposure range, the metatarsal side of the first metatarsal wedge joint was exposed medially and the metatarsal side of the third metatarsal wedge joint was exposed laterally; the peroneus longus tendon, Lisfranc plantar ligament and interosseous ligament were explored. X-ray films after the simulated operation showed satisfactory plate positions. (2) As for the preliminary clinical application, all patients were followed up for 6 to 14 months (mean, 11 months). At the last follow-up, the VAS score ranged from 0 to 1 (mean, 0.5), AOFAS score from 85 to 92 (mean, 89), Maryland score from 93 to 96 (mean, 95), and Kofoed score from 92 to 95 (mean, 94). There were no early complications such as fascial compartment syndrome, skin necrosis or infection. All fractures got united, with no complications like traumatic arthritis, muscle atrophy or screw loosening.Conclusion:Testified by the anatomical study, the plantar approach can be used to treat Lisfranc injury with poor dorsalis pedis soft tissue, metatarsal avulsion fracture or complicated multi-column lesions, leading to safe, effective and satisfactory clinical outcomes.

Plant metabolites are important for plant development and human health. Plants of celery (Apiumgraveolens L.) with different-colored petioles have been formed in the course of long-term evolution. However, the composition, content distribution, and mechanisms of accumulation of metabolites in different-colored petioles remain elusive. Using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), 1159 metabolites, including 100 lipids, 72 organic acids and derivatives, 83 phenylpropanoids and polyketides, and several alkaloids and terpenoids, were quantified in four celery cultivars, each with a different petiole color. There were significant differences in the types and contents of metabolites in celery with different-colored petioles, with the most striking difference between green celery and purple celery, followed by white celery and green celery. Annotated analysis of metabolic pathways showed that the metabolites of the different-colored petioles were significantly enriched in biosynthetic pathways such as anthocyanin, flavonoid, and chlorophyll pathways, suggesting that these metabolic pathways may play a key role in determining petiole color in celery. The content of chlorophyll in green celery was significantly higher than that in other celery cultivars, yellow celery was rich in carotenoids, and the content of anthocyanin in purple celery was significantly higher than that in the other celery cultivars. The color of the celery petioles was significantly correlated with the content of related metabolites. Among the four celery cultivars, the metabolites of the anthocyanin biosynthesis pathway were enriched in purple celery. The results of quantitative real-time polymerase chain reaction (qRT-PCR) suggested that the differential expression of the chalcone synthase (CHS) gene in the anthocyanin biosynthesis pathway might affect the biosynthesis of anthocyanin in celery. In addition, HPLC analysis revealed that cyanidin is the main pigment in purple celery. This study explored the differences in the types and contents of metabolites in celery cultivars with different-colored petioles and identified key substances for color formation. The results provide a theoretical basis and technical support for genetic improvement of celery petiole color.
Anthocyanins ; Apium/metabolism* ; Chlorophyll/metabolism* ; Color ; Gene Expression Regulation, Plant ; Humans ; Metabolomics ; Plant Proteins/genetics* ; Tandem Mass Spectrometry

ObjectiveTo analyze the production of Poria in 5 major producing areas and the problems through field investigation and literature review and thus to lay a foundation for healthy development of Poria industry. MethodEach production process of Poria in Anhui， Hubei， Hunan， Yunnan， and Sichuan was probed and related research was retrieved for the analysis. ResultSichuan， Guangxi， Guizhou， and other areas have been the emerging producers of Poria. However， a few Poria cocos lines were used and the variety breeding lagged behind. Different cultivation techniques were adopted in different production areas. For example， the "induction" method failed to be widely used. Moreover， the harvest time was mainly dependent on the market demand not matter it was suitable or not. Furthermore， steaming has replaced the traditional diaphoretic processing in the processing of this medicinal material in production areas. In addition， fumigation with sulphur was still used in the processing of Poria. ConclusionAn excellent variety is the key to the quality of Poria. Efforts should be made to strengthen the evaluation of the germplasm resources of P. cocos and variety breeding and standardize the processing in production areas， thereby ensuring the safety and effectiveness of Poria and the decoction pieces. In addition， the contradiction between artificial cultivation and woodland ecology should be fully coordinated to ensure the sustainable development of Poria industry in China.

ObjectiveHigh performance liquid chromatography （HPLC） was used to establish the specific chromatograms of Aurantii Fructus from different origins， and the quality variability of Aurantii Fructus from Sichuan was analyzed and evaluated by combining entropy weighting method and grey correlation method. MethodHPLC was performed on an Agilent Eclipse Plus C₁₈ column （4.6 mm×250 mm， 5 μm） with a gradient elution of methanol （A）-0.1% phosphoric acid aqueous solution as the mobile phase （0-12 min， 25%-33%A； 12-21 min， 33%-41%A； 21-30 min， 41%-42%A； 30-40 min， 42%-59%A； 40-53 min， 59%-72%A； 53-60 min， 72%A； 60-65 min， 72%-100%A； 65-70 min， 100%A； 70~71 min， 100%-25%A； 71-80 min， 25% A） at a flow rate of 1.0 mL·min^-1， the injection volume was 10 μL and the detection wavelength was 330 nm. Fifty batches of Aurantii Fructus samples from different origins （Sichuan， Chongqing， Jiangxi and Hunan） were tested， and the similarity evaluation software is used to generate characteristic profiles and compare them with control profile for peak identification， and then to evaluate the similarity of the samples. IBM SPSS 19.0 and SIMCA 14.1 were used to perform multivariate statistical analysis on the results of the samples， and then the entropy weighting method and grey correlation were used to calculate the overall quality score of samples from Sichuan. ResultHPLC specific chromatogram of Aurantii Fructus was established， and 14 common peaks were identified as eriocitrin， neoeriocitrin， narirutin， naringin， hesperidin， neohesperidin， meranzin hydrate， poncirin， meranzin， marmin， nobiletin， 3，3′，4′，5，6，7，8-heptamethoxyflavone， tangeretin and auraptene. And the similarities between the samples from Sichuan and the control chromatogram were all above 0.980. The samples could be classified into four categories according to their main origins by chemical pattern recognition， and the results of cluster analysis， principal component analysis and orthogonal partial least squares-discriminant analysis were all able to discriminate the samples of different main origins effectively. The comprehensive evaluation results of entropy weighting method combined with grey correlation showed that the quality of Aurantii Fructus from Sichuan varied greatly among different origins， and the quality of Aurantii Fructus from Sichuan was ranked as Bazhong>Luzhou>Chongqing>Neijiang. ConclusionIn this study， the characteristic mapping of Aurantii Fructus from Sichuan is established， and combined with the analytical methods of chemometrics and grey correlation， the quality of samples from different origins can be effectively differentiated， which can provide a reference for the comprehensive evaluation and control of the quality of Aurantii Fructus from Sichuan.

ObjectiveTo study the accumulation and absorption of dry matter and nutrients in Curcuma phaeocaulis from Sichuan， the origin of Curcumae Radix （tuber） and Curcumae Rhizoma （rhizome）， to explore the growth and development laws and nutrient demand characteristics of the medicinal species， and thus to lay a theoretical basis for rational fertilization. MethodThe plant growth indexes， dry matter accumulation， and nutrient content of C. phaeocaulis at different growth stages in Sichuan were measured and analyzed. ResultThis medicinal species featured the dominant growth of aboveground leaves and stems before October and growth of underground part （particularly the rhizomes and tubers） from October. During the whole growth period， the accumulation of nitrogen， phosphorus， potassium， calcium， magnesium， iron， manganese， zinc， and copper per plant was 2 450.31， 907.09， 3 171.18， 625.94， 493.38， 14.53， 2.24 ， 2.93， 0.46 mg， respectively， with the order of potassium > nitrogen > phosphorus > calcium > magnesium > iron > zinc > manganese > copper. ConclusionThe species needs sufficient potassium and nitrogen， appropriate amount of phosphorus， calcium， and magnesium， a small amount of iron， and very little zinc， manganese， and copper for growth， and potassium is particularly important. Nitrogen， phosphorus， and potassium fertilizers supply macroelements and nitrogen fertilizer should be supplemented at seedling stage， leafy stage， early rhizome expansion stage， tuber expansion stage， and the second expansion stage of rhizome. Phosphorus and potassium fertilizers should be applied at seedling stage， tuber expansion stage， secondary expansion stage of rhizome， and dry matter accumulation stage of tuber and rhizome. At seedling stage， leafy stage， early rhizome expansion stage， tuber expansion stage， and the second expansion stage of rhizome， a variety of medium and trace elements-containing foliar fertilizers should be used.

The anterior disc displacement (ADD) leads to temporomandibular joint osteoarthritis (TMJOA) and mandibular growth retardation in adolescents. To investigate the potential functional role of fibrocartilage stem cells (FCSCs) during the process, a surgical ADD-TMJOA mouse model was established. From 1 week after model generation, ADD mice exhibited aggravated mandibular growth retardation with osteoarthritis (OA)-like joint cartilage degeneration, manifesting with impaired chondrogenic differentiation and loss of subchondral bone homeostasis. Lineage tracing using Gli1-CreER⁺; Tm^fl/-mice and Sox9-CreER⁺;Tm^fl/-mice showed that ADD interfered with the chondrogenic capacity of Gli1⁺ FCSCs as well as osteogenic differentiation of Sox9⁺ lineage, mainly in the middle zone of TMJ cartilage. Then, a surgically induced disc reposition (DR) mouse model was generated. The inhibited FCSCs capacity was significantly alleviated by DR treatment in ADD mice. And both the ADD mice and adolescent ADD patients had significantly relieved OA phenotype and improved condylar growth after DR treatment. In conclusion, ADD-TMJOA leads to impaired chondrogenic progenitor capacity and osteogenesis differentiation of FCSCs lineage, resulting in cartilage degeneration and loss of subchondral bone homeostasis, finally causing TMJ growth retardation. DR at an early stage could significantly alleviate cartilage degeneration and restore TMJ cartilage growth potential.
Animals ; Mice ; Osteogenesis ; Zinc Finger Protein GLI1 ; Fibrocartilage ; Temporomandibular Joint ; Disease Models, Animal ; Osteoarthritis ; Stem Cells ; Growth Disorders