Objective To introduce an improved method of continuous intradermal suture,and to evaluate its clinical efficacy for the closure of surgical incisions.Methods Eighty-two patients were enrolled in this study,including 37 cases of nevus,10 cases of basal cell carcinoma,16 cases of sebaceous cyst,6 cases of lipoma,8 cases of seborrheic keratosis,3 cases of dermatofibroma and 2 cases of depressed scar.All the patients were managed by simple surgical excision with the shortest length of postoperative incisions being 0.8 cm and the longest length being 12 cm.An improved method of continuous intradermal suture was used for the closure of all the postoperative incisions.Specifically,an absorbable thread with a small triangle needle in both ends was inserted through and pulled out from the dermal layer at one side of the incision,and then inserted through and pulled out from the dermal layer at the opposite side of the incision,which was repeated until the incision was entirely closed.Results Among the 82 patients,80 achieved primary healing,and 2 developed erythematous painful swelling at the incision site 2 days after the operation,which disappeared after symptomatic treatment for 5 days.During 3-6 months of follow-up,the incisions closed leaving a flat and smooth surface in 78 patients,and proliferative scar formed in 4 patients,which was obviously improved after local injection with glucocorticoids.No disruption of incisions was observed.Conclusion The improved method of continuous intradermal suture can be applied to the closure of skin defects in the face,neck,trunk and extremities with a favorable healing outcome and cosmetic result.

A large number of studies have shown that long non-coding RNAs (IncRNAs) display abnormallity in organisms exposed to toxic chemicals,carcinogens and heavy metals.In this paper,the relationships between IncRNAs and microRNAs (miRNAs),the expressions of IncRNAs in organisms exposed to different exogenous toxic chemicals and the related toxicological mechanism are reviewed in order to provide reference for biological monitoring with IncRNAs in environmental toxicology.

AIM:To establish a fast , accurate and economical technique for culturing mouse pulmonary arte-riolar smooth muscle cells ( PASMCs ) , and to explore the effects of hypoxia on the proliferation and apoptosis of the PASMCs.METHODS:In sterile condition, the pulmonary artery was isolated from the male BALB/c mice by digesting with collagenase I, and the cells were cultured in fetal bovine serum-coated flask.Centrifugal procedure was not used dur-ing the cell passage .The cell morphology was observed under an inverted phase-contrast microscope .α-Smooth muscle ac-tin was identified by immunocytochemistry and immunofluorescence .The effects of hypoxia on the proliferation and apopto-sis of the PASMCs were detected by CCK-8 assay and TUNEL assay .RESULTS:PASMCs were identified by the methods of immunocytochemistry , immunofluorescence staining and observation of morphology .Unlike the rat PASMCs with typical subcultured peak-vally pattern, the mouse PASMCs showed a lot different without a peak-vally pattern.The cells could be subcultured after 5 d to 7 d and there was 3 to 5 generations depending on the activity of the cells .CCK-8 assay demonstra-ted that the A values of PASMCs exposed to hypoxia increased after 24 h ( P<0.05) as compared with normoxia .TUNEL result showed that the apoptotic index of the PASMCs in hypoxia decreased after 24 h (P<0.05).CONCLUSION:This technique for obtaining cultured mouse PASMCs is simple , fast, accurate and economical .The digestion time is easy to control.Hypoxia promotes the proliferation and inhibits the apoptosis of PASMCs .

Objective To investigate autoantibody against endothelin receptor type A (ENRA-Ab) in patients with systemic lupus erythematosus associated pulmonary arterial hypertension (SLE-PAH).The possibility of autoantibody-mediated pathogenesis in the development of SLE-PAH has also been explored.Methods ENRA-Ab in the serum of SLE-PAH and controls were detected by using a human ETRA epitope peptide-based ELISA.The clinical relevance of ENRA-Ab in SLE-PAH was analyzed.Proliferation of vascular smooth muscle cells (SMCs) and permeability of endothelial cells in vitro under the stimulation of polyclonal ENRA-Ab IgG were assessed.The expressions of PAH-related markers, i.e., 5-HTT, PDGFR-b, VEGF-A and PDGF-B were measured by qPCR.The effect of ENRA-Ab in vivo was also determined in a suboptimaldose monocrotaline-induced model with the assessment of right ventricle hypertrophy index and pathology parameters.Independent t-test, Tukey-Kramer test of variance analysis and Pearson' s correlation analysis were used for statistical analysis.Results ENRA-Abs was presented in a higher occurrence in SLE-PAH (35/85,41％) compared with controls (0/60;0, 13/80, 16％).There was a significant correlation between ENRA-Ab and echocardiograph estimated pulmonary arterial systolic pressure (r=0.392, P=0.002) in SLE-PAH.ENRA-Ab could promote SMCs proliferation, disrupt endothelial barrier and up-regulate PAH-related markers expression,which could be blocked in the presence of ETR antagonist.ENRA-Ab aggravated right ventricle hypertrophy and vascular remodeling in vivo.Conclusion ENRA-Ab is a new biomarker, in SLE-PAH, which may mediate PAH development in SLE.

OBJECTIVE: To observe the protective effect of Huaxia shallot preparation on human umbilical vein endothelial cell (HUVEC) injury induced by oxidized low density lipoprotein (Ox-LDL) in vitro. METHODS: Ox-LDL was prepared and identified, and HUVECs were cultured. After 2-hour intervention of different drugs and 24-hour following intervention of Ox-LDL, the number of HUVECs was observed by phase contrast optical microscope and the activity of the HUVECs was observed by methyl thiazolyl tetrazolium (MTT) technique. Superoxide dismutase (SOD) activity and nitric oxide (NO) content were assayed by respective kit. The protein expressions and mRNA levels of peroxisome proliferators activated receptor gamma(PPAR-gamma) and endothelial nitric oxide synthase (eNOS) were measured by western blot technique and reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Ox-LDL could increase the apoptosis rate of the HUVECs and decrease the NO release as compared with the blank control group (P<0.05). These effects induced by Ox-LDL were all significantly inhibited by Huaxia shallot preparation. It could up-regulate the protein expressions and mRNA levels of PPAR-gamma and eNOS significantly (P<0.05). Huaxia shallot preparation could decrease the apoptosis rate of the HUVECs. CONCLUSION: Ox-LDL may be involved in the initiation and progression of atherosclerosis by injuring the endothelial cells directly and may cause the endothelial dysfunction. Huaxia shallot preparation can protect against Ox-LDL induced endothelial cell injury by up-regulating the protein expressions and mRNA levels of PPAR-gamma and eNOS. It suggests that Huaxia shallot preparation may play a role in the prevention and treatment of cardiovascular disease.

Objective To compare the performance of four commercial anti-dsDNA antibody assays,i.e,BioPlex 2200 (BioPlex),Farr radioimmunoassay (Farr),MESACUP DNA-Ⅱ TEST ds [MBL-enzyme linked immunosorbent assay (ELISA)] and Anti-dsDNA-NcX ELISA (IgG) (EURO-ELISA),Antoantibodies Profile Assay Kit (HOB-Chemiluminescent Immunoassay) in disease activity assessment of systemic lupus erythematosus (SLE).Methods SLE patients (n=119) as well as healthy controls (n=200) and disease controls (n=100) were recruited and their serum anti-dsDNA antibodies were detected by BioPlex,Farr,MBL-ELISA,EURO-ELISA,and a standard Crithidia luciliae indirect immunofluorescence test (CLIFT).The consistency between above four methods to CLIFT was analyzed.The correlation of anti-dsDNA antibody level of these four methods to SLE disease activity was assessed.All data analyses were performed with Statistical product and service solutions (SPSS) 16.0 (SPSS.Inc) and GraphPad Prism 4.0.3 (GraphPad).Unless otherwise specified,all data in this study were expressed as mean±standard deviation.Cut-off values of the anti-dsDNA quantification methods were set by the manufacturers.Chi square and kappa coefficients were adopted to assess the agreement determination and correlation analysis between anti-dsDNA level and SLE disease activity (SLEDAI).Receiver-operator characteristic (ROC) curve analysis was used to compare the specificity and sensitivity of the anti-dsDNA assays.Student's t test was adopted for the comparison of anti-dsDNA levels by different methods between SLE and SLE+LN groups.A p value small than 0.05 was considered statistically significant.Results Using cut-off values set by the manufacturers,BioPlex demonstrated the highest overall agreement with CLIFT,while MBL-ELISA and EURO-ELISA showed the highest positive agreement with CLIFT.Disease activity correlation analysis showed that SLEDAI score correlated poorly with anti-dsDNA level in Farr assay,but strongly with the other three assays.Bioplex had a better performance in terms of SLE activity index corelation (r=0.297 6,P=0.001 2).Moreover,anti-dsDNA level differed in SLE patients with renal lupus nephritis in BioPlex assay (P=0.026 8),but not in the other assays.In ROC curve analysis,BioPlex showed the largest area under the curve (AUC) over other assays.Conclusion Bio Plex assay has better sensitivity and specificity than Farr,MBL-ELISA and EURO-ELISA and correlates well with SLE disease activity.

OBJECTIVETo evaluate the effect of co-culture system of bone marrow mesenchymal stem cells (BMSC) and vascular endothelial cells (EC) on osteogenesis.

METHODSBMSC were isolated by whole bone marrow centrifugal adherent method. Then BMSC were induced into EC with induced medium. Co-culture system in different proportions of BMSC and EC (10:0, 10:1, 8:2, 7:3, 5:5, 3:7, 2:8, 1:10, 0:10) were further evaluated. The cell growth level of BMSC was examined. The CD44 expression of BMSC and von willebrand factor (vWF) expression of vascular EC were examined by immunofluorescence. Furthermore, calcium nodules exhibited by alizarin red staining, alkaline phosphatase activity, and the expression of osteogenic genes by reverse transcription-quantitative PCR (RT-qPCR) were observed to validate the osteogenesis of co-culture system.

RESULTSThe growth curve of P3 passage of BMSC demonstrated the doubling time of BMSC was 39.9 h. The positive specific markers of BMSC and EC showed efficient induction. Although the calcium nodules ratio of the co-culture [group 7:3 (19.0 ± 3.0) and group 5:5 (20.8 ± 2.9)] was not significantly different (P > 0.05), but higher than that of other co-culture groups with a significant difference (P < 0.01). Alkaline phosphatase activity was increased with prolonged induction of osteogenic medium. While alkaline phosphatase activity of group 10:0 (16.84 ± 0.82), group 10:1 (15.86 ± 3.10), group 8:2 (16.37 ± 1.33), group 7:3 (17.99 ± 1.98), and group 5:5 (17.49 ± 0.87) did not show significant difference after osteogenic induction for 7 days (P > 0.05), but significantly higher than that of other co-culture groups (P < 0.05). The co-culture ratio of 7:3 (33.74 ± 0.99) was slightly higher than that of 5:5 (31.09 ± 0.87), but significantly higher than that of other groups (P < 0.01). Moreover, the osteocalcin (OCN) and runt-related transcription factor 2 (RUNX2) expression of group 7:3 was significantly higher than that of other groups.

CONCLUSIONSThe EC that derived from BMSC can promote the BMSC differentiate into osteoblasts. The co-culture system of BMSC and EC with the ratio of 7:3 increases the alkaline phosphatase activity and facilitates the expression of osteogenic genes.

Alkaline Phosphatase ; metabolism ; Bone Marrow Cells ; cytology ; physiology ; Cell Culture Techniques ; Cell Differentiation ; Cell Proliferation ; Coculture Techniques ; Core Binding Factor Alpha 1 Subunit ; metabolism ; Endothelial Cells ; cytology ; physiology ; Humans ; Hyaluronan Receptors ; metabolism ; Mesenchymal Stromal Cells ; cytology ; physiology ; Osteoblasts ; cytology ; Osteocalcin ; metabolism ; Osteogenesis ; physiology ; Time Factors ; von Willebrand Factor ; metabolism

OBJECTIVE： To investigate the proportion of 8 kinds of ginsenoside to ginsenoside Rg1 in Panax ginseng and the regularity of growth year in Jilin province， and to provide reference for the identification of growth year. METHODS： The samples of garden ginseng， wild-cultivated ginseng and wild ginseng were collected from different growth years （3-30 years） in Jilin province. The contents of 8 ginsenoside （ginsenoside Rg1， Re， Rf， Rb1， Rc， Rb2， Rb3， Rd） in P. ginseng were determined by HPLC. The contents of saponins as well as the proportion of 8 kinds of ginsenoside to ginsenoside Rg1 were calculated； the relationship of the proportion with growth year was investigated. RESULTS： As the increase of growth year， the proportion of 8 kinds of ginsenosides in garden ginseng to ginsenoside Rg1 as well as that of ginsenoside Re， Rb1， Rc， Rd to ginsenoside Rg1 were decreased gradally （P＜0.001）； the proportion of ginsenoside Re to ginsenoside Rg1 in wild-cultivated ginseng decreased first and then increased（P＜0.001）； the proportion of 8 kinds of ginsenosides to ginsenoside Rg1 as well as the proportion of ginsenoside Re and Rb1 to ginsenoside Rg1 were increased gradually in wild ginseng （P＜0.001）； the proportion of ginsenoside Rf， Rb3 to ginsenoside Rg1 in garden ginseng， wild-cultivated ginseng and wild ginseng had no significant difference（P＞0.05）. CONCLUSIONS： Garden ginseng， wild-cultivated ginseng and wild ginseng contain 8 kinds of ginsenosides. The growth year can be predicted preliminarily according to the proportion of ginsenoside Rg1 and ginsenoside Re， Rb1 to ginsenoside Rg1.

Methanol has become an attractive substrate for the biomanufacturing industry due to its abundant supply and low cost. The biotransformation of methanol to value-added chemicals using microbial cell factories has the advantages of green process, mild conditions and diversified products. These advantages may expand the product chain based on methanol and alleviate the current problem of biomanufacturing, which is competing with people for food. Elucidating the pathways involving methanol oxidation, formaldehyde assimilation and dissimilation in different natural methylotrophs is essential for subsequent genetic engineering modification, and is more conducive to the construction of novel non-natural methylotrophs. This review discusses the current status of research on methanol metabolic pathways in methylotrophs, and presents recent advances and challenges in natural and synthetic methylotrophs and their applications in methanol bioconversion.
Humans ; Methanol/metabolism* ; Metabolic Engineering ; Metabolic Networks and Pathways ; Biotransformation