In vitro culture expansion impairs chondrogenic differentiation and the therapeutic effect of mesenchymal stem cells by regulating the unfolded protein response

In vitro expansion of mesenchymal stem cells (MSCs) has been implicated in loss of multipotency, leading to impaired chondrogenic potential and an eventual therapeutic effect, as reported in our previous study. However, the precise regulatory mechanism is still unclear. Here, we demonstrate that endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) were involved in transformation of MSCs induced by in vitro culture based on the comparative profiling of in vitro cultured bone marrow MSCs at passage 3 (P3 BMSCs) vs. fresh P0 BMSCs by microarray analysis. Indeed, RT-PCR and Western blot analysis showed significantly lower expression levels of three key UPR-related molecules, ATF4, ATF6 and XBP1, in P3 BMSCs than P0 BMSCs. Further, we found that UPR suppression by 4-phenylbutyrate (4-PBA) reduced the chondrogenic potential of P0 BMSCs and further cartilage regeneration. Conversely, UPR induction by tunicamycin (TM) enhanced the chondrogenic differentiation of P3 BMSCs and the therapeutic effect on cartilage repair. Thus, the decline in the chondrogenic potential of stem cells after in vitro culture and expansion may be due to changes in ER stress and the UPR pathway.     

454 pyrosequencing analysis on microbial diversity of an expanded granular sludge bed reactor treating high NaCl and nitrate concentration wastewater

The treatment of high-salinity, high-nitrate wastewater was investigated in a single expanded granular sludge bed reactor. Complete denitrification was achieved when nitrate concentration was as high as 6,000 mg N/L and the salinity of influent reached 11% NaCl at liquid up-flow velocity of 3.0 m/h, hydraulic retention time of, 24 h and the C/N molar ratio of 2.0. Furthermore, 454-pyrosequencing technology was used to analyze archaea bacterial diversity under high salinity and high nitrate conditions. Results showed that the total number of effective sequences was 5749 consisting of 5678 bacterial sequences and 71 archaea sequences after denoising and filtering out chimeras, which could be affiliated to 5 phylogenetic groups, including Proteobacteria, Firmicutes, Euryarchaeota, Crenarchaeota and unclassified phylum. Although Proteobacteria was the dominant microbial population, two archaea phylogenetic groups-Crenarchaeota and Euryarchaeota were observed in this study.     

Residue Val237 is critical for the enantioselectivity of Penicillium expansum lipase

The shape of the hydrophobic tunnel leading to the active site of Penicillium expansum lipase (PEL) was redesigned by single-point mutations, in order to better understand enzyme enantioselectivity towards naproxen. A variant with a valine-to-glycine substitution at residue 237 exhibited almost no enantioselectivity (E = 1.1) compared with that (E = 104) of wild-type PEL. The function of the residue, Val237, in the hydrophobic tunnel was further analyzed by site-directed mutagenesis. For each of these variants a significant decrease of enantioselectivity (E < 7) was observed compared with that of wild-type enzyme. Further docking result showed that Val237 plays the most important role in stabilizing the correct orientation of (R)-naproxen. Overall, these results indicate that the residue Val237 is the key amino acid residue maintaining the enantioselectivity of the lipase.     

Heme Oxygenase-2 Suppress TNF-α and IL6 Expression via TLR4/MyD88-Dependent Signaling Pathway in Mouse Cerebral Vascular Endothelial Cells

Heme oxygenase (HO) represents an intrinsic antiinflammatory system based on its ability to inhibit expression of proinflammatory cytokines. The constitutive isoform heme oxygenase-2 (HO-2) has high expression and activity in cerebral microvascular endothelial cells (CMVEC). This study was undertaken to evaluate the role of HO-2 in regulation of TLR4/MyD88-dependent signaling and to study the effect of HO-2 on the expression and secretion of the proinflammatory cytokines tumor necrosis factor α (TNF-α) and Interleukin-6 (IL6) in CMVEC. HO-2 short hairpin RNA (shRNA) and HO-2 overexpression plasmids were used to observe the effect of HO-2 on proinflammatory cytokines in CMVEC in vitro, and the results showed that the messenger RNA (mRNA) and protein levels of TNF-α and IL6 were increased and decreased, respectively, compared with control groups. LPS-stimulated TNF-α and IL6 mRNA and protein were also reduced in CMVEC treated with an inhibitor of TLR4 signaling, CLI-095, or HO-2 overexpression. CLI-095 and HO-2 overexpression both reduced TLR4 expression in CMVEC, and HO-2 shRNA blocked these effects of CLI-095. CLI-095 and HO-2 overexpression potently suppressed TLR4/MyD88-dependent proinflammatory cytokine expression in CMVEC. These results suggest that HO-2 plays an important role in protecting CMVEC against cytokine-mediated inflammation.     

Studies on Chemical Constituents of Essential Oil from Leaves of Jiang-Zhang

Jiang-Zhang is a Physiological type of Cinnamomum porrectum (Roxb.) Kosterm. The essential oil of leaves of Jiang-Zhang can be extracted by steam distillation, with yields of 0.5–0.8%. It contained dtral (α-,β-citral) 64.11% and can be used in aromatic and medicinal industries. We used the methods of GC, IR, GC/MS/DS and prepared derivative and 47 chemical constituents were identified, as follows: α-thujene (0.06 %), α-pinene (2.42%), camphene (1.26%), sabinene (0.21%),β- pinene (1.38%) myrcene(0.38%), α-phellandrene (0.32%),△3-carene (0.01%), p-cymene(0.21%),α-limonene (1.57%), 1 8-cineole (0.82 % ), β-phellandrene (0.10 %), cis-linalool oxide(0.07 % ), linalool (8.43 % ) epicamphor(0.26 % ), camphor(1.10 %), borneol( 1.07 % ), β-citral (neral) (28.28 % ), geraniol (0.25%), nerol (0.47%), α-citral (geranial) (35.83%), methyl citronellate (0.12%), n-undecane (0.18), safrole (0.02), methyl geranate (0.23%), geranyl formate (0.09%), α-copaene (0.12%), trans-methyl cinnamate (0.02%), n-dodecane (0.20), β-elemene (0.12%), caryophyllene (4.67%), α-guaiene (0.04), β-guaiene (0.06%), β-selinene (0.97%), azulene (0.38%), β-cubebene (0.30%), n-pentadecane (0.05%), β-gurjunene (0.04 %), epi-β-santalene (0.46 %), aremophliene (0.05 % ), alloaromadendrene (0.03 % ), α-elemene (0.26%), trans-β-farnesene (0.04%), (z)-β-farnesene (0.30%), r-elemene (0.03%), β-bisaboiol(0.41%), cedrol (0.16%), respectively.     

从中指骨长度推算身高的研究

作者对近年在华南地区收集的,已知生前身高的汉族成年男性骨骼的中指骨近节、中节进行了测量。用直线回归方程、多元回归方程对从中指骨长度推算身高进行了研究。并用50例国人骨骼标本对这些推算身高的方法作了检验。结果表明,中指骨与四肢大型长骨,以及从中指骨长度推算身高的直线回归方程与多元回归方程,对推算身高的价值都是相同的。