Afferent Pathways ; anatomy & histology ; Animals ; Female ; Ganglia, Spinal ; anatomy & histology ; Neurons, Afferent ; Rats ; Rats, Sprague-Dawley ; Urinary Bladder ; innervation

Objective To study interleukin-1 receptor(IL-1R) and connexin(Cx36) gene expression following Mg 2+-free-induced seizures in cultured developing neuron. Methods Rat embryo cortical neurons cultured for 6 and 17 days were exposed to Mg 2+-free media to induce seizure. At different time after Mg 2+-free treatment, real-time RT-PCR was used to detect IL-1R and Cx36 mRNA expression. Results 1. IL-1R mRNA expression transiently decreased after Mg 2+-free treatment in neurons cultured for 6 and 17 days in vitro. Then the levels of IL-1R mRNA expression recovered in neurons cultured for 6 days, but IL-1R mRNA expression were increased in neurons cultured for 17 days compared with control group and the peak was at 24 hours. 2. In neurons cultured for 6 days in vitro, Cx36 mRNA expression increased after Mg 2+-free treatment compared with control group, the peak was at 24 hours. But in neurons cultured for 17 days in vitro, Cx36 mRNA expression decreased at 6 hours after Mg 2+-free treatment compared with control group, the peak was at 24 hours. Conclusions IL-1R mRNA and Cx36 mRNA expression following Mg 2+-free-induced seizures are different between the neurons cultured for 6 and 17 days in vitro. This is possibly related to the different neuron injury between 6 and 17 days in vitro following seizures.

The marine biological source of mineral drugs recorded in Chinese Pharmacopoeia (2010 version) mainly including pearl, nacre, clam shell, common oyster shell, ark shell, cuttle bone, and sea-ear shell are widely used in clinical. Calcium carbonate and a small amount of protein are the main components in this type of drugs. In this paper, a systematical and comparable study were carried out by determination of calcium carbonate by EDTA titration method, the crystal of calcium carbonate by X-Ray powder diffraction and the total amino acids (TAAs) of the hydrolyzed samples by ultraviolet spectrophotometry method. As a result, the crystal structure is calcite for common oyster shell, mixture of calcite and aragonite for nacre and sea-ear shell, aragonite for the other drugs. The content of calcium carbonate ranged from 86% to 96%. Cuttle bone has the highest amount of TAAs among the seven drugs which reached 1.7% while clam shell has the lowest content of 0.16% on average. In conclusion, an effective method was developed for the quality control of marine mineral drugs by comprehensive analysis of calcium carbonate and TAAs in the seven marine mineral drugs.
Amino Acids ; analysis ; chemistry ; Animal Shells ; chemistry ; Animals ; Calcium Carbonate ; analysis ; chemistry ; Crystallization ; Edetic Acid ; chemistry ; Mollusca ; chemistry ; classification ; Pharmaceutical Preparations ; analysis ; chemistry ; standards ; Quality Control ; Reproducibility of Results ; Seawater ; Species Specificity ; Spectrophotometry, Ultraviolet ; X-Ray Diffraction

Background Studies demonstrated that human amniotic epithelial cells (AECs) have some characteristics of embryonic stem cells and they were used to re-establish the surface of eyes. Human AECs may serve as new seed cells in tissue engineering for corneal epithelium reconstitution in the future. Objective The present study was to investigate the application value of human amniotic epithelium cells transfected by lentiviral vectormediated enhanced green fluorescent protein (EGFP) gene as new seed cell source for engineering the corneal surfacelayer. Methods Lentiviral vector carrying the objective gene EGFP was transfected into human amniotic epithelial cells (pLenti6/V5-DEST),and the transient expression of the transgene in the human amniotic epithelial cells was observed under the fluorescence microscope. Flow cytometry was used to detect the positive expression rates of EGFP in transfected cells. The transfected human amniotic epithelial cells were seeded onto the fresh corneal stromal surface of New Zealand white rabbit and cultured in vitro. The stem cell deficiency ( SCD ) models were established by cutting off the limbus of cornea in 20 eyes of New Zealand white rabbits, and the model rabbits were then divided into 2 groups randomly. The transplanted grafts carrying the pLenti6/V5-DEST-EGFP gene-transferred human amniotic epithelium cells were regarded as the pLenti6/V5-DEST-EGFP group, and the corneal stroma graft without any epithelial cell served as the control group. The opacity of stroma and corneal conjunctivalization and vascularization were observed daily. The rabbits' eyes were extracted one month after operation. The expression of EGFP in the cornea was detected under the fluorescence microscope, and the expression of CK8, CK18 and CK12 in cornea was detected by immunohistochemical staining. Results The shape of the transferred human amniotic epithelial cells resembled normal human amniotic epithelial cells. 48 hours after the transient transfection of EGFP presented with the highest expression level throughout the observation duration, with a positive expression rate of EGFP of 61.5% ,showing significant differences in comparison with that of 12 ( 5.24% ) , 24 ( 38.27% ) or 96 ( 39. 10% ) hours ( P ＜0. 05) post-transfection; but no obvious difference was found in the positive rate of transiently transfected EGFP between 48 hours and 72 hours ( 58.36% ) ( P＞0. 05 ). Six cornea grafts were clear in 1 month and two corneas were rejected during the observation period in the pLenti6/V5-DEST-EGFP group. A few new blood vessels were seen around the graft. Ten corneas of the control group became opaque and cloudy with new blood vessels growth around the grafts. Imunohistochemistry revealed the positive expressions of CK8, CK1 8 and CK12 in the corneal epithelial layer in the pLenti6/V5-DEST-EGFP group. However,the expression of CK12 was absent in the control group. Conclusion Human amniotic epithelium cells transfected with the pLenti6/V5-DEST-EGFP gene is a new and ideal feed cell type to reconstruct the corneal surface layer. Lentivirus is a relatively safe gene transfection vector.

The aim of this paper was to investigate the effect of terpene penetration enhancers on membrane fluidity and membrane potential using HaCaT keratinocytes, and study the potential mechanisms of these terpene compounds using as natural transdermal penetration enhancer. Six terpene compounds, namely menthol, limonene, 1,8-cineole, menthone, terpinen-4-ol and pulegone, were chosen in this study on account of their good penetration-enhancement activities. The cytotoxicity of these terpene compounds was measured using an MTT assay. The fluorescence recovery after photobleaching (FRAP) technique was employed to measure the change of membrane fluidity of HaCaT cells. The flow cytometer was used to study the alteration of membrane fluidity of HaCaT cells, and investigate the effect of terpene compounds on intracellular Ca2+. It was found that 6 terpene compounds possessed low cytotoxicity in comparison to the well-established and standard penetration enhancer azone. Those terpene compounds could significantly enhance HaCaT cells membrane fluidity and decrease HaCaT cells membrane potentials. Meanwhile, after treated with various terpene compounds, the Ca2(+)-ATPase activity and intracellular Ca2+ of HaCaT cells was decreased significantly. Terpene penetration enhancers perhaps changed the membrane fluidity and potentials of HaCaT cells by altering the Ca2+ balance of the cell inside and outside, resulting in the low skin permeability to increase the drug transdermal absorption.
Cell Line ; Drugs, Chinese Herbal ; pharmacokinetics ; Humans ; Keratinocytes ; drug effects ; metabolism ; Membrane Fluidity ; drug effects ; Skin Absorption ; drug effects ; Terpenes ; pharmacokinetics

Glomerular hypertrophy is the main pathological characteristic in the early stage of diabetic nephropathy (DN), and its regulatory mechanism is closely related to mammalian target of rapamycin (mTOR) signaling pathway activity. mTOR includes mTOR complex 1 (mTORC1) and mTOR complex 2(mTORC2), in which, the upstream pathway of mTORC1 is phosphatidylinositol-3-kinase (PI3K)/serine-threonine kinase(Akt)/adenosine monophosphate activated protein kinase(AMPK), and the representative signaling molecules in the downstream pathway of mTORC1 are 4E-binding proteins(4EBP) and phosphoprotein 70 S6Kinase(p70S6K). Some Chinese herbal extracts could improve cell proliferation via intervening the expressions of the key molecules in the upstream or downstream of PIK/Akt/mTOR signaling pathway in vivo. As for glomerular mesangial cells(MC) and podocyte, mTOR plays an important role in regulating glomerular inherent cells, including adjusting cell cycle, energy metabolism and matrix protein synthesis. Rapamycin, the inhibitor of mTOR, could suppress glomerular inherent cell hypertrophy, cell proliferation, glomerular basement membrane (GBM) thickening and mesangial matrix deposition in model rats with DN. Some Chinese herbal extracts could alleviate glomerular lesions by intervening mTOR signaling pathway activity in renal tissue of DN animal models or in renal inherent cells in vivo and in vitro.
Animals ; Diabetic Nephropathies ; drug therapy ; enzymology ; genetics ; pathology ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hypertrophy ; drug therapy ; enzymology ; genetics ; pathology ; Kidney Glomerulus ; drug effects ; metabolism ; pathology ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; genetics ; metabolism

ObjectiveThis work is aimed to investigate the possible association of dendritic cell immunoreceptor (DCIR) with rheumatoid arthritis (RA) susceptibility in Chinese Han population.Methods A total of 523 patients with RA and 510 healthy controls were genotyped for single-nucleotide polymorphism (SNP) rs2377422 and rs10840759.Association analyses were performed on the whole data set and on RA subsets based on the status of anti-cyclic citrullinated peptide antibody (CCP) in RA patients.Finally,we carried out the association analysis of rs2377422 with DCIR mRNA expression in RA patients.Statistical analysis used in this study included X2 test,Logistic regression,and Mann-Whitney U test.ResultsDCIR rs2377422 was found significantly associated with RA(allele analysis: OR 1.26; 95％CI 1.06～1.51,P=0.005; genotype analysis CC vs TT+TC: OR 1.34; 95％CI 1.18～2.06,P=0.004).Following stratification for anti-CCP antibody status,association of ra2377422 with anti-CCP-positive RA was observed(allele analysis: OR 1.22,95％CI 0.99～1.48,P=0.055).In contrast,the SNP rs2377422 was found specifically susceptible to anti-CCP-negative RA(allele analysis: OR 1.46; 95％CI 1.10～1.93,P=0.0091; genotype analysis CC vs TT+TC: OR 1.58;95％CI 1.01～2.47,P=0.043),despite loss of power in the analysis.DCIR gene transcription quantification analysis further proved the dominant effect of rs2480256 CC genotype on DCIR mRNA expression levels in RA patients (CC vs TT+TC: 0.429±0.069 vs 0.238±-0.023,U=1861,P=0.0015).ConclusionThe study provides evidence for the association between DCIR rs2377422 and RA,particularly with anti-CCP-negative RA in Chinese Han populations.

Background The diagnosis of central serous chorioretinopathy (CSC) is mainly dependent onfluorescine fundus angiography (FFA). However, the combination of optical coherence topography (OCT) with FFA offers a new approach to the research of the pathogenesis of CSC. Objective This clinical study was designed to study the combined application of the FFA and OCT for the research of the pathogenesis of central serous chorioretinopathy (CSC). Methods Forty-four eyes of 44 patients with CSC were included in this study with 36 cases of males and 8 cases of female. The patients were aged 39.3 ± 5.3 years and the visual acuity was 0. 64 ±0. 27. FFA and OCT examinations were performed in all patients and the FFA images were imported into the Topcon 3D OCT 1000 device to locate the conformity of OCT lesions with the leakages of FFA. The neuroepithelial layer thickness at the fovea and the height of the neuroepithelial layer detachment were measured using 3-D OCT. Results OCT showed serous REP detachment in 34 eyes (77.3%) and rough surfaces of RPE in 10 eyes (22. 7% ). In thirtyfour eyes with RPE detachment, the OCT lesions and FFA leakage spots conformed to the same locations in 31 eyes, but the other three eyes did not. The mean foveal neuroepithelial thickness was (138.5±19.4) μm in CSC eyes and that of normal eyes was ( 131.35±5. 01 ) μm ,showing a significant difference between them( t=0. 39 ,P＞0. 05 ). The mean height of neuroepithelial detachment was (263.3 ± 126.7 ) μm in CSC eyes. Conclusion RPE detachment occurs in CSC eyes and further induces macular neuroepithelial detachment. Leakage lesion of fluorescine corresponds to RPE detachment. CSC without RPE detachment may be related to the increase in RPE permeability. OCT can accurately measure the thickness of the macular neuroepithelial layer and the height of the neuroepithelial detachment.

Abstract: Objective To analyze the effect of nutrition package on the nutritional status and prevalence of children in rural areas of Hainan Province, and provide scientific basis and suggestions for further improving the nutritional and health status of children in this region. Methods　Four cities and counties were randomly selected as the intervention group, and four cities and counties matched with the intervention group in terms of population, economy, social culture, maternal and child health work foundation of township health centers, physical nutrition and health status were selected as the control group.With the combination of monitoring and prospective cohort study, infants in the intervention group and the control group were studied from June 1, 2020, and they were intervened for 12 months with supplementary food nutrition package. Before and after intervention, the nutrition and health status of infants aged 6-24 months in the intervention group and the control group were investigated to evaluate the nutritional and health effects of supplementary food nutrition package for infants aged 6-24 months in rural Hainan Province. Results　A total of 999 infants were investigated, including 427 in the intervention group and 572 in the control group. After 12 months of nutritional intervention, there was no significant difference in weight-for-age Z-score (WAZ) and height-for-age Z-score (HAZ) and weight-for-height Z-score (WHZ) between the intervention group and the control group (P>0.05). The rate of emaciation of the intervention group was 1.64%, which was significantly lower than 3.67% of the control group (P<0.05). There were no significant differences in the rate of growth retardation (2.81% and 3.32%, respectively) and underweight (0.47% and 1.92%, respectively) between the intervention group and the control group (P>0.05). The rate of respiratory infection and diarrhea in the intervention group were 9.13% and 1.17%, which were significantly lower than corresponding 23.25% and 3.15% in the control group (P<0.05). The hemoglobin of the intervention group and the control group were 117.24 g/L and 114.51 g/L respectively, and the rates of anemia were 11.11% and 22.84% respectively, the differences were statistically significant (P<0.05). Conclusions　The intervention of nutrition package in rural areas of Hainan Province has achieved the expected results, and supplementary food nutrition package has reduced the incidence of malnutrition and respiratory infection and diarrhea in recent two weeks in infants and anemia to a certain extent. We should attach great importance to the supplementary nutrition package for right-age children and promote the growth and health of children in rural areas through supplementary nutrition package, and continuously improve the nutrition and health level of children in Hainan Province.

ObjectiveTo explore the mechanism of extracorporeal shock wave (ESW) in treating osteogenic disorders and the ideal energy level. MethodsAfter success in marrow aspiration from patients' iliac crest, hMSCs were isolated by Percoll density gradient centrifugation and cultured in Dulbecco's modified Eagle's medium in a 5% CO2 and 37 ℃ incubator. Optimal ESW dose was determined by MTT of kinase-marked cytobiology. After hMSCs were exposed to ESW, their morphocytologic change, rate of adherence and doubling time were observed with IPCM. Enzyme cytochemistry reaction for the activity of alkaline phosphatase was also examined. ResultsESW of 5 kV and 100 times could increase cells' viability and proliferation (P<0.01), but higher than 7 kV would inhibit them. Rate of adherence of hMSCs in exposure group of passage 5 reached to 61.54%, which was significantly different from control group(P<0.05). Compared with control group, the MSCs' doubling time was short for 1.72 d (P<0.05). The curve of normal alkaline phosphatase activity of hMSCs was like type S, but ESW shortened its latent period, and promoted its peak time, which was significantly different from control group.ConclusionESW of 5 kV and 100 times can optimally promote the proliferation and activity of osteogen of hMSCs in vitro.