BACKGROUND:Combining seed cells with 3D bioprinting technology enables the specific construction of various tissues and organs to meet the demands of tissue repair.However,further research is needed on the promotion of angiogenesis in damaged tissues. OBJECTIVE:By cultivating a 3D scaffold structure of methacrylated gelatin loaded with fibroblasts,obtaining the supernatant,and mixing it in different proportions with a complete culture medium to simulate the cellular microenvironment during tissue repair,this study aimed to explore the role of various cellular microenvironments in promoting angiogenesis in endothelial cells. METHODS:A methacrylated gelatin scaffold structure loaded with fibroblasts was prepared using an extrusion-based 3D bioprinting process.Hydrogel scaffold extract was prepared and mixed with a complete culture medium in ratios of 1:1,1:2,and 1:4 to obtain conditioned medium.Mouse embryonic fibroblasts BALB3T3 and human umbilical vein endothelial cells were co-cultured with complete medium(control group)and hydrogel scaffold extract,respectively.Cell proliferation was assessed using the CCK-8 assay and cell viability was analyzed using live/dead staining.Three kinds of conditioned medium and complete medium(control group)were used to co-culture with human umbilical vein endothelial cells for tube formulation assay,vascular genetic testing,and immunofluorescence staining of CD31. RESULTS AND CONCLUSION:(1)Scanning electron microscopy revealed that the methacrylated gelatin scaffold exhibited a porous structure,and rheological results demonstrated excellent mechanical properties of the hydrogel.CCK-8 assay and live/dead cell staining showed that the hydrogel scaffold extract had no obvious cytotoxicity.(2)Tube formulation assay indicated that the hydrogel showed the total length of cell tubules in 1:1 conditioned medium group was smaller than that in the control group(P<0.05).There were no statistical differences among the four groups in the number of vascular branches formed by endothelial cells(P>0.05).(3)qRT-PCR results showed that for vascular endothelial growth factor mRNA expression,the 1:2 conditioned medium group was lower than the 1:1 conditioned medium group on day 1(P<0.01).On day 3,the expression level of vascular endothelial growth factor in the 1:2 conditioned medium group was higher than that in the control group(P<0.01).On day 5,the cytokine expression level in the 1:2 conditioned medium group was significantly higher than that in the other three groups(P<0.01 or P<0.000 1).The expression in the 1:1 conditioned medium group was significantly lower than that in the other three groups(P<0.05 or P<0.01).On day 1,the expression level of basic fibroblast growth factor in the 1:1 conditioned medium group was significantly higher than that in the control group and 1:4 conditioned medium group(P<0.01,P<0.05).The expression was higher in the 1:2 conditioned medium group than that in the control group(P<0.05).On day 3,the expression levels of cytokines in the 1:4 conditioned medium group was higher than that in the control group(P<0.05).(4)On day 3,the expression of CD31 in the 1:2 conditioned medium group was higher than that in the control group and the 1:4 conditioned medium group(P<0.05).(5)The results indicate that the resulting conditioned media can simulate the microenvironment of vascular regeneration after tissue damage,promoting the vascularization process of endothelial cells.The best promotion of vascularization in endothelial cells was observed when the ratio of supernatant to complete culture medium was 1:2.

Objective:To evaluate effect of diammonium glycyrrhizinate(DG)on lipopolysaccharide(LPS)-induced TLR4/NF-κB inflammatory signaling pathway in BV2 microglial cells and to explore its potential regulatory mechanisms on ameliorating neu-roinflammation.Methods:BV2 microglia injury model was induced by LPS and treated with 20 and 60 μmol/L DG.MTS was used to determine vitality of cells.NO level secreted by cells was detected by Griess.Inflammatory factors TNF-α,IL-6,and IL-1β levels in cell supernatant were measured by ELISA.TNF-α,IL-6,IL-1β,TLR4 and MyD88 mRNA levels were detected by RT-qPCR.Western blot was used to determine expressions of TLR4,MyD88,NF-κB,p-NF-κB,IκB-α and p-IκB-α proteins of cells.Results:Compared with control group,LPS-treated BV2 microglia had significantly lower viability(P<0.05),significantly higher NO secretion(P<0.05),significantly up-regulation levels of inflammatory factors TNF-α,IL-6 and IL-1β(P<0.05),and significantly higher mRNA levels of TNF-α,IL-6,IL-1β,TLR4 and MyD88(P<0.05),TLR4,MyD88,NF-κB,p-NF-κB,IκB-α,and p-IκB-α protein expressions were significantly increased(P<0.05).Compared with LPS group,BV2 microglia intervened with different concentrations of DG had significantly higher viability(P<0.05),significantly lower NO secretion(P<0.05),significantly lower TNF-α,IL-6 and IL-1β inflammatory factors levels(P<0.05),significantly lower mRNA levels of TNF-α,IL-6,IL-1β,TLR4 and MyD88(P<0.05),and TLR4,MyD88,p-NF-κB and p-IκB-α protein expressions were significantly reduced(P<0.05).Conclusion:DG can inhibit LPS-induced neuroinflammatory responses in microglia,whose underlying mechanism is suppression of TLR4/NF-κB signaling pathway.

Dental erosion is characterized by progressively destroyed teeth, which has no relation to bacteria but to chemicals. Some internal factors, such as gastroesophageal reflux induced by bulimia, anorexia, gastrointestinal diseases, or drugs, and external factors, such as diet, drugs, and occupational acid exposure, are considered promotive factors for this disease. This article presents a patient suffering from severe dental erosion in the whole dentition, especially in the maxillary teeth, due to gastroesophageal reflux induced by glucocorticoid therapy for optic neuritis. This article discusses the mechanism between optic neuritis glucocorticoid therapy and dental erosion.
Humans ; Glucocorticoids/therapeutic use* ; Tooth Erosion/therapy* ; Gastroesophageal Reflux/complications*

A case of IgG4-related hypertrophic pachymeningitis was reported. The patient was an elderly female, with the course of disease more than 8 years. Clinical manifestations included recurrent headache, vision and hearing loss, exophthalmos and thyroid dysfunction. Finally, she was diagnosed as IgG4-related disease and IgG4-related hypertrophic pachymeningitis by PET-CT and dural biopsy. After treatment with methylprednisolone and mycophenolate mofetil, the patient′s clinical symptoms improved.

ObjectiveTo analyze the quality changes of Platycladi Semen before and after the deterioration of moth-eaten and rancidity during storage. MethodFour types samples of Platycladi Semen， including normal， moth-eaten， oxidative rancidity and hydrolytic rancidity， were determined for volatile components， odor， and taste based on headspace solid phase microextraction/gas chromatography-mass spectrometry （HS-SPME/GC-MS） and electronic sensory techniques such as electronic nose and electronic tongue. Volatile components were identified by searching the database and manual comparison， the odor and taste were determined by the response values of the electronic nose and electronic tongue sensors， and the difference between samples before and after deterioration was studied by multivariate statistical analysis. ResultA total of 85 compounds were identified in Platycladi Semen samples. Compared with the normal samples， the number of volatile compounds in samples after hydrolytic rancidity decreased by 5， the number of volatile compounds in samples after moth-eaten and oxidative rancidity increased by 1 and 21， respectively. Aldehydes and acids accounted for majority of types. Among them， the contents of N-hexanoic acid， hexanal and propionic acid in the samples of oxidative rancidity reached 11.49%， 10.21% and 7.52%， which became the key indicators of rancidity. There was significant variance among the odor components corresponding to W1W， W2W and W1S sensors by electronic nose analysis. It was indicated that the value of sourness in deteriorated samples generally increased by mean of electronic tongue analysis. Compared with normal samples， the moth-eaten samples had changed slightly and rancidity samples had changed significantly especially oxidative rancidity samples of volatile components， odor and taste by multivariate statistical analysis. ConclusionIn terms of Platycladi Semen， the oxidative rancidity caused by nature storage for 12 months has the greatest impact on the quality. Therefore， it should be mainly to prevent oxidative rancidity to ensure the quality of Platycladi Semen.

Although traditional treatment for Graves′ disease(GD) displays some effects, it is imperative to explore new treatment methods. Based on the pathogenesis of GD, biologic agents developed by consumption of B lymphocytes and acting on thyroid stimulating hormone receptor(TSHR), such as monoclonal antibodies, TSHR antagonists and epitopes, can provide more options for patients with GD, and some new drugs are expected to be put into clinical practice. By restoring the damaged immune system and maintaining normal thyroid function continuously, it can avoid the disadvantages of conventional therapies such as long-term treatment, induction or aggravation of Graves ophthalmopathy, permanent hypothyroidism, and other complications. Preliminary experience suggests that thermotherapy is effective and safe for patients with refractory GD. In addition, the traditional Chinese medicine improves the symptoms and thyroid function of GD patients.The emergence of new therapeutic modalities and techniques will provide new approaches for the future treatment of GD and help clinicians to make the best decision.

Objective:To establish and evaluate a predictive model for recurrence risk of Graves′ disease after antithyroid drugs(ATD) withdrawal.Methods:Among 308 patients with newly onset Graves′ disease taking ATD from 2012 to 2019, 170 patients who completed follow-up were enrolled and divided into relapse and remission groups according to whether hyperthyroidism reoccurred within 2 years after ATD withdrawal to establish the discovery cohort. An internal validation cohort was constructed by repeating the sampling with bootstrap. Cox regression analysis was used to screen risk factors and establish a predictive model, named Graves′ Recurrence Evaluation System(GRES). The differentiation and accuracy of GRES model were evaluated and compared with the GREAT score.Results:Of 170 patients, 90 Graves′ disease cases relapsed within 2 years after ATD withdrawal. According to Cox regression analysis, family history of Graves′ disease, younger age(<30 years), grade Ⅱ-Ⅲ goiter, high level of TRAb(≥13 IU/L), large thyroid volume(≥26.4 cm 3) and low 25(OH) D(<14.7 ng/mL) were included in the predictive model: PI=0.672×family history+ 0.405×age+ 0.491×severity of goiter+ 0.808×TRAb+ 1.423×thyroid volume+ 0.579×25(OH) D. PI≥1.449 was associated with a higher risk of recurrence after drug withdrawal. The GRES model has good prediction in assessing Graves′ disease relapse within 2 years after ATD withdrawal and better than GREAT score. Conclusion:GRES model can be used to evaluate the recurrence risk within 2 years for patients with newly onset Graves′ disease after ATD withdrawal, and facilitate clinicians to reasonably select treatment modalities in order to improve the remission rate.

Pulmonary fibrosis (PF) is a chronic, progressive, fatal interstitial lung disease with limited available therapeutic strategies. We recently reported that the protein kinase glycogen synthase kinase-3

Vitamin D has been found to produce therapeutic effects on obesity-associated insulin resistance and dyslipidemia through its potent anti-inflammatory activity, but the precise immunomodulatory mechanism remains poorly understood. In the present study we found that 1,25-dihydroxyvitamin D [1,25(OH)D], the biologically active form of vitamin D, significantly attenuated monosodium glutamate (MSG)-induced obesity and insulin resistance as indicated by body weight reduction, oral glucose tolerance improvement, and a glucose infusion rate increase as detected with hyperinsulinemic-euglycemic clamp. Moreover, 1,25(OH)D not only restored pancreatic islet functions but also improved lipid metabolism in insulin-targeted tissues. The protective effects of 1,25(OH)D on glycolipid metabolism were attributed to its ability to inhibit an obesity-activated inflammatory response in insulin secretory and targeted tissues, as indicated by reduced infiltration of macrophages in pancreas islets and adipose tissue while enhancing the expression of in liver tissue, which was accompanied by increased infiltration of Treg cells in immune organs such as spleen and lymph node as well as in insulin-targeted tissues such as liver, adipose, and muscle. Together, our findings suggest that 1,25(OH)D serves as a beneficial immunomodulator for the prevention and treatment of obesity or metabolic syndrome through its anti-inflammatory effects.

OBJECTIVETo investigate the relationship between serum/follicular fluid fetuin B levels and outcome offertilization (IVF).

METHODSInfertility women (28 with low fertilization rates, 44 with normal fertilization rates) receiving IVF in Women's Hospital of Zhejiang University School of Medicine during June and December 2016 were enrolled in the study. Serum/follicular fluid fetuin B levels were measured with ELISA method. Correlations of serum and follicular fetuin B level with fertilization outcome of IVF were analyzed with Pearson correlation coefficient and receiver operating characteristic (ROC) curve.

RESULTSA positive correlation between serum fetuin B and follicular fluid fetuin B levels was observed (=0.675,<0.01). Both serum and follicular fluid fetuin B levels in women with low fertilization rates of IVF were lower than those in women with normal fertilization rates[(6.09±1.31) μg/mL vs. (7.13±1.47) μg/mL,=3.050,<0.05; (5.13±0.96)μg/mL vs. (6.22±1.33) μg/mL,=3.755,<0.01]. ROC analysis showed that the area under curve (AUC) of serum fetuin B level in predicting fertilization rate was 0.832 (95%:0.729-0.934,<0.01), and 6.08 μg/mL could be used as cut-off value.

CONCLUSIONSSerum fetuin B level is correlated with follicular fluid fetuin B level, and it may be used for predicting the fertilization outcome of IVF.