Objective To investigate the feasibility of animal model of the reconstruction of right ventricular outflow tract in rats. Methods A total of 15 female Sprague-Dawley (SD) rats underwent right ventricular outflow tract reconstruction surgery. Before the operation, the collagen scaffolds were treated with g 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride chemistry (EDC), and seeded with human bone marrow stem cells (h-MSCs). Three days after the surgery, 3 rats were randomly sacrificed to evaluate the transmural resection of right ventricular outflow tract. One or 3 months later, other 3 rats at each timepoint were sacrificed, stained with Masson’s Trichrome to observe the degradation of scaffold. Furthermore, 4 weeks after the surgery, 4 rats were sacrificed and the hearts were sliced. Anti-human mitochondria staining was used to identify the survival of seeding cells. Results The transmural resection of right ventricular outflow tract was feasible in rats at an acceptable mortality (13.3%). After EDC treatment, the degradation rate of collagen scaffold was extended greatly. The seeding cells were detected by anti-mitochandria immunofluorescent staining in all patches 4 weeks after the operation. Conclusion Rat model of right ventricular outflow tract reconstruction could be a stable, reliable and economical screening model for engineered heart tissue research.

AIM:To observe the concentration of RBP4 and IL-6 in vitreous of proliferative diabetic retinopathy (PDR). METHODS: A total of 65 patients (66 eyes) were enrolled in Department of Ophthalmology, Renmin Hospital of Wuhan University from February 2017 to July 2017 with the informed consent. The patients were divided into PDR group (23 cases) and NPDR group (16 cases). Twenty- six patients without diabetic mellitus (DM) served as control group. The demography was matched among the groups, but the course of DM, the blood glucose level and the HbA1c level were elevated in the PDR group and the NPDR group (all P<0.05). Vitreous samples were collected during the procedure of vitrectomy. RBP4,IL-6,TNF-α concentrations in vitreous specimens were detected by ELISA. The differences of vitreous RBP4, IL-6 and TNF-α in various groups were statistically analyzed by ANOVA, respectively. The correlations between RBP4 and IL - 6, TNF - α were calculated by Pearson correlation analysis. RESULTS:The concentration of RBP4 in PDR group,the NPDR group and control group were 13.68士2.66, 11 03士1 12,10.45士1.17μ g/Ml, and the concentration of IL-6 were 56.0士10.27, 20.92士5.77, 10.26士1.91pg/Ml. RBP4 and IL- 6 concentrations were elevated in PDR group compared with NPDR group and control group, with significant difference among three groups (F = 12. 135, 161.167; P < 0. 01). IL - 6 concentrations in vitreous increased in the NPDR group in comparison with control group(P<0.05). RBP4 concentrations had no significant difference between the NPDR group and the group(P>0 05). Pearson correlation coefficient was significant positive between RBP4 concentration and IL - 6 concentration(r=0.606,P=0.001). CONCLUSION: RBP4 is probability involved in the inflammation pathogenesis of PDR. These results indicate that RBP4 could be a new target for the diagnosis and treatment of PDR.

OBJECTIVETo investigate the effect of MicroRNA-3963(miR-3963) on the adipogenic differentiation of mouse bone-derived mesenchymal stem cells(MSC).

METHODSMSCs were isolated from C57BL/6 mice bone fragment and transfected with miR-3963 mimic, miR-3963 inhibitor and negative control. The expression of miR-3963 and transfection efficiency were detected by q-PCR. These transfected cells were induced to adipocytes and stained with oil red O after 14 days culture. q-PCR and Western blot were used to detect the expression of adipogenic differentiation marker genes C/EBPα and PPARγ at transcriptional level and protein level.

RESULTSThe results of q-PCR revealed that miR-3963 expression level was up-regulated after transfection with miR-3963 mimic (P<0.0001), and down-regulated after transfection with miR-3963 inhibitor (P<0.0001). After oil red staining, overexpression of miR-3963 in MSCs could promote the formation of lipid droplet. The q-PCR and Western blot analyses showed the significant increase of expression of adipogenic marker genes C/EBPα and PPARγ in MSC transfected with miR-3963 mimic. Additionally, compared with the control group, miR-3963 inhibitor could decrease adipogenic differentiation of MSC.

CONCLUSIONmiR-3963 can regulate and promote adipogenic differentiation of mouse bone-derived MSC.

Field surveys and literatures show that Polygonati Rhizome (Huangjing) was firstly recorded in Shen Nong&s Herbal Classic, and widely used as a medicinal and edible plant. It has a long history of cultivation, and the researches on chemistry have made some progress. The future development is prospected on health market. But the Polygonati Rhizome industry has faced a lot of problems, including the resource depletion, unstable quality, low-tech in cultivation and germplasm confusion, unclear of functional composition, decentralized, small scaled and primary processing products. The suggestion for sustainable development are listed below. First, the relevant researches should focused on material basis and biological mechanism of core effects. To speed up the selection and breeding of improved varieties, ensure the supply of high-quality seedlings and eliminate the unauthentic species are the most important measures. Secondly, to strengthen the conservation and rational use of wild resources, break through the key technologies of high-quality artificial cultivation on light regulation, site control, density control and precision harvesting are also very important. Thirdly, to reveal the toxicity-reducing-and-efficacy-enhancing mechanism of processing, optimize the parameters and setup the standard operating procedure are indispensable. Fourthly, that full advantage of the root, leaf, flower and fruit resources should be strengthened for enlarged health products based on the development of exact functional factors. Above all, Polygonati Rhizome could be a growing market in the future driven by the technological innovation, cultural creativity, integration of three industries, brand strategy and internet+technique.

Human cytosolic sulfotransferase 2A1 (SULT2A1) is an important phase II metabolic enzyme. The detection of SULT2A1 is helpful for the functional characterization of SULT2A1 and diagnosis of its related diseases. However, due to the overlapping substrate specificity among members of the sulfotransferase family, it is difficult to develop a probe substrate for selective detection of SULT2A1. In the present study, through characterization of the sulfation of series of bufadienolides, arenobufagin (AB) was proved as a potential probe substrate for SULT2A1 with high sensitivity and specificity. Subsequently, the sulfation of AB was characterized by experimental and molecular docking studies. The sulfate-conjugated metabolite was identified as AB-3-sulfate. The sulfation of AB displayed a high selectivity for SULT2A1 which was confirmed by reaction phenotyping assays. The sulfation of AB by human liver cytosols and recombinant SULT2A1 both obeyed Michaelis-Menten kinetics, with similar kinetic parameters. Molecular docking was performed to understand the interaction between AB and SULT2A1, in which the lack of interaction with Met-137 and Tyr-238 of SULT2A1 made it possible to eliminate substrate inhibition of AB sulfation. Finally, the probe was successfully used to determine the activity of SULT2A1 and its isoenzymes in tissue preparations of human and laboratory animals.

·Age - related macular degeneration ( ARMD ) and Stargardt's macular dystrophy ( SMD ) are two kinds of degenerative retinal diseases that respectively lead to irreversible vision loss of the elderly and juvenile population. However, the severe visual impairment in dry ARMD and SMD remains untreatable. In recent years, with the advancement of stem cell technology, stem cell-derived RPE cell transplantation therapy of retinal degeneration has become new research hotspot and direction. This article reviewed the progress of stem cell based approaches for treating retinal degenerative diseases and discussed the prospect and challenges in this field.

As medicine and food plants, the market demand of Zamthoxyli Pericarpium is extensive, and the sanshool in Zamthoxyli Pericarpium is getting attention gradually. However, because of the low content and not establishing rating evaluation method for sanshool and lack of standards, the research on substance basis and physiological activity is necessary. This article reviewed the extraction, separation, purification and biological activities of sanshoolin Zamthoxyli Pericarpium in recent years to provide development and utilization of Zamthoxyli Pericarpium.

Objective To summarize and review the comparative results of the isotope dilution mass spectrometry in Cholesterol Reference Method Laboratory Network (CRMLN)project of the Centers for Disease Control and Prevention (CDC)of the United States in order to provide quality controls for determination of blood lipid.Methods The isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS)methods for determination of serum total cholesterol (TC)and triglycerides (TG)were developed to participate in the comparison of CRMLN.The survey was conducted every three months before 2016 and every half year from 2016.Four kinds of reference materials with two parallel tubes for each material and each tube in duplicate were determined in every survey.At least two certified reference materials used as quality control samples were analyzed simultaneously in each determination.Results In the 15 comparisons the CV of the TC determination method in our laboratory was 0.43％ while the CV of all the participated laboratories in CRMLN was 0.42％.The bias from the overall mean value in our laboratory was 0.22％ while the bias from the CDC target values was 0.58％.The CV of the TG determination method in 15 tests of our laboratory was 0.62％ and the bias was-0.98％ from the overall mean value and-0.80％ from the target values of CDC.Among the 60 results for comparison,98％ (59/60)of CV in the TC determination met with CDC requirement for precision (CV≤ 1％),and 70％ of bias (42/60) of the results met with CDC requirement for accuracy (bias ≤ 1％).For the 60 results in comparison of TG determination,92％ (55/60)of bias of the results met with the accuracy requirement of CDC (bias ≤2.55％).Conclusion In CRMLN comparison the results of TC and TG determined by ID-LC/MS/MS method were consistent with the values which were certified by CDC and determined by other network laboratories.The comparative surveys may play an important role in the standardization of lipid determination,and should be expected to provide experiences and technical supports in the comparative plan for reference measurement in our country.

This study evaluated the effects of sodium hypochlorite (NaOC1) with different concentrations and exposure time on the structural,compositional and mechanical properties of human dentin in vitro.Sixty dentin slabs were obtained from freshly extracted premolars,randomly distributed into four groups (n=15),and treated with 1％,5％,10％ NaOC1 and distilled water (control group),respectively,for a total of 60 min.Attenuated total reflection infrared (ATR-IR) spectroscopy,Raman spectroscopy and X-ray diffraction (XRD) were carried out before,10 min and 60 min after the treatment.Scanning electron microscopy (SEM) and flexural strength test were conducted as well.The results showed that dentins experienced morphological alterations in the NaOC1 groups,but not in the control group.Two-way repeated-measures analysis of variance revealed that the carbonate:mineral ratio (C:M),Raman relative intensity (RRI),a-axis,c-axis length and full width at half maximum (FWHM) with the increase of time and concentration in the NaOC1 groups were not significantly different from those in the control group (P＞0.05).Nevertheless,the mineral:matrix ratio (M:M) increased and the flexural strength declined with the increase of concentration and the extension of time in the NaOC1 groups (P＜0.05).Additionally,it was found that the M:M and the flexural strength remained unchanged after 1％ NaOCl treatment (P＞0.05),and the morphology changes were unnoticeable within 10 min in 1％ NaOC1 group.These results indicated that NaOC1 has no significant effects on the inorganic mineral of human dentin;but it undermines and eliminates the organic content concentration-and time-dependently,which in turn influences the flexural strength and toughness of dentins.In addition,an irrigation of 1％NaOCl within 10 min can minimize the effects of NaOC1 on the structural and mechanical properties of dentin during root canal treatment.

Diabetic retinopathy(DR) and diabetic nephropathy(DN) are the most common complications of diabetes, and the main causes of death and disability of diabetes. Clinical reports showed that Fufang Xueshuantong capsule(FXT) had effective curative effect on DR and DN, but there was no report on the distribution of chemical compounds of FXT in beagle dog eyes and kidneys. In this study, FXT was given by gavage administration in Beagle dogs for 3 days, and then their eyeballs and kidneys were taken. The chemical compounds in beagle dog eyes and kidneys were detected by HPLC LTQ-Orbitrap technology. Furthermore, by comparing with the data from retrieving literature and references, the chemical compounds were identified by the accurate mass, retention time (tR), and MS/MS. Fourteen and 19 types of notoginsenosides were respectively identified in eyeballs and kidneys in this study, and these results could lay foundation for clarifying the effective ingredients of FXT in treatment of DR and DN.