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Taiwania cryptomerioides is an endangered relict conifer species native to East Asia with a variety of practical applications. To contribute to its conservation and sustainable utilization, its complete chloroplast genome was assembled from Illumina sequencing reads. The circular genome is 131,427 bp in size with a biased nucleotide composition (33.5% A, 17.5% C, 17.2% G & 31.8% T). It encodes a panel of 114 gene species, including 83 protein-coding genes (PCGs) and 31 tRNAs. Gene duplication was detected for three tRNA gene species (trnI-CAU, trnQ-UUG & trnV-GAC). Besides, 15 gene species (atpF, ndhA, ndhB, petB, petD, rpl16, rpl2, rpoC1, rps16, trnA-UGC, trnG-UCC, trnI-GAU, trnK-UUU, trnL-UAA & trnV-UAC) possess a single intron, and another one (ycf3) has a couple of introns. Phylogenetic analysis suggests that T. cryptomerioides and T. flousiana are closely related to each other within the family Cupressaceae. [ABSTRACT FROM AUTHOR]

A possible correlation between liver cancer pain and the hypoxia-inducible factor (HIF)-1 and vascular endothelial growth factor (VEGF) expression levels was examined. From January, 2015 to January, 2016, 30 patients suffering from liver cancer with pain, 30 patients with liver cancer without pain and 30 hepatitis patients with pain were enrolled in the study. Pain level was evaluated by visual analogue scale (VAS), the expression levels of HIF-1 and VEGF mRNA were determined by RT-PCR and the expression levels of HIF-1 and VEGF proteins were examined by ELISA. Before intervention, the VAS in the hepatitis group was significantly higher than that of the liver cancer pain group. However, after intervention the VAS in the two groups was reduced. HIF-1 and VEGF mRNA expression levels in the liver cancer pain group were significantly higher than those in the liver cancer group before and after intervention. The expression levels of HIF-1 and VEGF mRNA in the hepatitis group were the lowest. The expression levels of HIF-1 and VEGF mRNA in the liver cancer pain group considerably increased after intervention. The expression levels of HIF-1 and VEGF mRNA in the other two groups showed no changes before or after intervention. Before and after the intervention, VAS in the liver cancer pain group was positively correlated to the expression levels of HIF-1 and VEGF. Thus, pain occurrence and the pain level in liver cancer patients were correlated with the expression levels of HIF-1 and VEGF. As the regular three-step medicine analgesic ladder is ineffective in these cases, verification of HIF-1 and VEGF expression levels may be considered the new target for pain release.

Objective: To explore whether a tissue-engineered construct composed of autogenous endothelial cells, osteoblasts and a new bioresorbable nano-hydroxyapatite/recombinant human-like collagen/polylactic acid (nHA/RHLC/PLA) would enhance bone regeneration and repair femoral head defects in canine models., Methods: The bone marrow stem cells (BMSCs) were isolated from bone marrow of canine ilium and cultured in Dulbecco's modified eagle medium:nutrient mixture F-12 culture media for 1 week and the second-generation BMSCs were further induced by osteogenic medium (1×10(-8) mol/L dexamethasone, 10 mmol/L B-sodium glycerophosphate and 50 µg/ml vitamin C) and by endothelial cell grow medium (vascular endothelial growth factor and basic fibroblast growth factor) for 14 days in vitro. Thus BMSCs were induced into ECs and OBs. After the second passage, cells were digested and collected.And cell density was adjusted to 1.0×10(6)/ml.The cells and nHA/RHLC/PLA scaffold were co-cultured for 2-4 hours then nHA/RHLC/PLA scaffold composites prepared. Cavity defects of 8 mm in diameter and 10 mm in height were made in femoral heads.The nHA/RHLC/PLA scaffold composited with ECs and osteoblasts (OBs) (group A) and composited with OBs (group B) were inserted into different defects while cell-free nHA/RHLC/PLA scaffold served as controls (group C). New bone formation and defect repair were evaluated at 3 and 6 months by radiographic examination, histology and bone histomorphometry., Results: New bone formation was evident as early as 3 months in groups A, B and C.At 6 months, abundant bone tissue within defects was observed in group A. The control animals with cell-free scaffold showed less bone formation at both timepoints.The scaffold of nHA/RHLC/PLA was degraded and absorbed gradually with the formation of new bone tissues.Histology and bone histomorphometry further revealed significantly increased trabecular bones in group A compared with groups B and C at 6 months postimplantation (P < 0.01)., Conclusion: More abundant new bone tissue may be found in the bone defect areas implanted with osteoblast-endotheliocyte composite than osteoblasts composite and scaffold materials only.ECs and osteoblasts derived from BMSC are ideal seed cells for repairing femoral head defects.

Background: Vulnerable plaques play an important role in the onset of sudden cardiac events and strokes. How to stabilize vulnerable plaques is still a challenge to medical science. Alprostadil is a biologically active substance with strong activity on vessel. Our study assessed the stabilizing effects of an alprostadil liposome microsphere preparation (ALMP) on vulnerable plaques in the brachiocephalic artery of apolipoprotein E (Apo E) knockout mice., Methods: Seventy-two male Apo E-knockout mice were fed a high-fat diet beginning at eight weeks of age. At week 17, they were divided randomly into groups for treatment with a high dose (3.6 µg×kg(-1)×d(-1)) or low dose (1.8 µg×kg(-1)×d(-1)) of an ALMP, or 0.2 ml/d normal saline (control group). The drug was administered using a micro-capsule pump. Twenty weeks after drug administration, pathological changes in the vulnerable plaques within the brachiocephalic artery were assessed, and levels of anti-mouse monocyte/macrophage monoclonal antibody (MOMA-2) and superoxide anions in the plaques were detected using immunofluorescence. The soluble intercellular adhesion molecule-1 (ICAM-1) expression was measured by ELISA, and the expression of matrix metalloproteinase-9 (MMP-9) and CD40 mRNA was measured using RT-PCR. Thrombospindin-1 (TSP-1) expression was detected using Western blotting., Results: Compared with the control group, ALMP treatment significantly reduced the plaque area in the brachiocephalic artery (P < 0.01), significantly lowered the contents of the lipid core (P < 0.01), significantly reduced the number of ruptured fibrous caps (P < 0.05), and increased the thickness of the fibrous cap and significantly reduced the incidence of intra-plaque hemorrhage (P < 0.05). ALMP treatment significantly reduced the expression of MOMA-2, superoxide anion, MMP-9, ICAM-1 and CD40 in the plaques (P < 0.01), decreased plasma ICAM-1 expression (P < 0.01), and increased the expression of TSP-1., Conclusions: Treatment with ALMP can stabilize vulnerable plaques by inhibiting inflammation.

Objective: To establish nonalcoholic fatty liver disease (NAFLD) in young rats, and to investigate the metabolic characteristics of these rats., Methods: Fifteen male and fifteen female SD rats of 3 weeks old were randomly divided into three groups, normal group (N), 20% high fat group (HF1) and 30% high fat group (HF2). All the rats were fed under Specific pathogen Free (SPF) condition for 6 weeks and executed at the end of the 6th week. Body length and weight of each rat as well as their liver weight were measured for calculating Liver Index (LI). ALT, AST, TG, TC, INS, Glu and HOMA-IR in the blood were measured. Liver tissue homogenate was prepared for detecting TG level. The liver section was stained with HE and oil red. The expression of SPEBP-1 and leptin in liver was detected by immunostaining., Results: The typical pathological change of NAFLD was found in the rats of HF groups. In HF2 group, no rats died during the experiment and the degree of fat degeneration is homogeneous. Comparing with those in N group, TC (mmol/L), liver TG (mmol/L) and ALT levels in HF2 group were significantly elevated (2.50+/-0.39 vs 1.82+/-0.43, P less than 0.01; 25.38+/-13.29 vs 12.09+/-9.59, P less than 0.01 and 69.80+/-18.22 vs 48.00+/-10.45, P less than 0.01, respectively). Comparing with those in N group, TG level in HF1 group was significantly decreased (0.17+/-0.10 vs 0.32+/-0.12, P less than 0.05), Glu level in HF1 group was significantly elevated (12.33+/-3.48 vs 8.13+/-2.53, P less than 0.05). There were no significant difference between the results of AST, INS and HOMA-IR among the groups. The expression level of SREBP-1 and leptin increased in HF groups., Conclusion: NAFLD can be induced by 30% high-fat feeding for 6 weeks in young rats, high-fat feeding induces the expression of SREBP-1 and leptin expression and fat synthesis.

Aim: To construct the expression vectors of procalcitonin (PCT), prepare polyclonal antibodies (pAbs) and monoclonal antibodies (mAbs) against PCT and identify their specific biological activity., Methods: The recombinant expression plasmids of pGEX-4T-1-PCT and PET-32a-PCT were constructed using thyroid carcinoma cell line (TT cell) cDNA as template. The fusion protein of His-PCT was expressed in E.coli and used as immunogen. The specificity of antiserum against human PCT was characterized by ELISA, Western blot and indirect immunofluorescence. The mAbs against human PCT were identified by Western blot and indirect immunofluorescence., Results: The recombinant expression plasmids of pGEX-4T-1-PCT and PET-32a-PCT were constructed and the fusion protein of His-PCT was expressed and purified. The antiserum against human PCT was prepared and the titer detected by ELISA was 1:256 000. The pAb specifically recognized the recombinant human PCT. Eight hybridoma cell lines secreting specific mAbs against PCT were established. The mAbs recognized the recombinant human PCT and four of them recognized the native PCT of TT cytoplasm in immunofluorescent assay., Conclusion: The successful preparation of polyclonal and monoclonal antibodies against human PCT is beneficial to further research into the pathological and physiological functions of PCT in severe bacterial infection and sepsis.