Objective To evaluate the effectiveness of ultrafiltration technology in endotoxin removal from purified recombinant MUC1-MBP fusion protein (MUC1-MBP)and to demonstrate the effect of ultrafiltration on endotoxin removal.Methods CM Sepharose FF weak cation exchange (CM)(CM group), CM combined with Phenyl Sepharose 6 FF exchange (C6)(CM+C6 group),CM combined with ultrafiltration (CM+ultrafiltration group), and CM combined with C6 and ultrafiltration (CM+C6+ultrafiltration group)were used to purify the MUC1-MBP from E.coli. and remove endotoxin;the expression level of endotoxin was detected by Chromogenic End-point Tachypleus Amebocyte Lysate.Results There was a single band at the expected molecular weight of 62 000 by SDS-PAGE analysis.and the purity>96% by Quantity One analysis.The endotoxin levels in CM group and CM +C6 group were quite high and there was no significant difference between two groups (P>0.05 );the endotoxin level in CM+ultrafiltration group was significantly lower than that in CM group, and there was significant difference (P<0.01);the endotoxin level in CM+C6+ultrafiltration group was significantly decreased compared with CM+C6 group (P<0.01);there was no significant differences of endotoxin level between CM+ultrafiltration group and CM +C6+ultrafiltration group (P>0.05 ). Conclusion The effects of CM or CM combined with C6 on endotoxin removal are quite poor, especially C6;CM combined with ultrafiltration are quite effective on endotoxin removal,and ultrafiltration plays an important role in endotoxin removal.

Objective To analyze the risk factors associated with multiple organ dysfunction syndrome (MODS) in children after cardiopulmonary bypass (CPB).Methods Between Jan 2001 and Dec 2010,1 899 patients undergoing open heart surgery were reviewed retrospectively according to the presence or absence of MODS.Univariate and multivariate logistic regression analysis were used to identify the risk factors.Results Thirty-five patients (1.84％) developed MODS,the mortality for MODS was 51.43％ (18/35).Univariate risk factors included complex congenital heart disease,perioperative unexpected events,CPB time,aortic cross-clamping time,mechanical ventilation time,and postoperative spsis.Multivariate logistic regression analysis identified that complex congenital heart disease,perioperative unexpected events,CPB time ＞ 180 min,postoperative spsis were risk factors.Conclusion The results suggest that the patients with MODS risk factors described above need more careful peri and post operative surveillance and preventive management.

OBJECTIVETo investigate the effect of hypoxia on the proliferation and expressions of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in keratinocytes obtained from oral lichen planus (OLP) lesions.

METHODSHypoxia environment was induced by a airtight incubator. Five groups were included, normoxia control group, hypoxia control group (12, 24, 36, 48 h). The effect of different treatment time of hypoxia on cellular proliferation was determined with cell counting kit-8 (CCK-8). The mRNA and protein expressions of HIF-1α, VEGF and MMP-9 were analyzed respectively by quantitative real-time polymerase chain reaction with SYBR Green I and Western blotting.

RESULTSThe growth activity of keratinocytes obtained from OLP lesions in the hypoxia groups (0.340±0.002, 0.415±0.006, 0.546±0.006) was reduced than that in control (0.431±0.001, 0.620±0.004, 1.022±0.005) (P < 0.01). The mRNA levels of VEGF (2.087±0.291, 3.189±0.573, 5.402±0.563) and MMP-9 (2.936±0.500, 4.083±0.300, 6.374±0.858) were elevated by hypoxia (P < 0.05). The protein levels of HIF- 1α (0.414±0.093, 0.751±0.056, 0.875±0.040), VEGF (0.393±0.046, 0.557±0.078, 0.767±0.045) and MMP-9 (0.250±0.053, 0.384±0.038, 0.611±0.092) were all remarkably elevated by hypoxia (P < 0.05). However, hypoxia had no effect on HIF-1α mRNA expression. The mRNA expression of HIF-1α after hypoxia exposure for 36 h (1.412±0.094) and 48 h (1.417±0.446) was higher than that of control group, however, there was no significant difference. A positive correlation was noted between HIF-1α and VEGF in protein level (r = 0.905, P = 0.000), and the same correlation found between HIF-1α and MMP-9 (r = 0.881, P = 0.000).

CONCLUSIONSHypoxia conditions may inhibit the proliferation of keratinocytes obtained from OLP lesions. Hypoxia conditions can promote the protein expressions of HIF-1α and both the mRNA and protein expression of VEGF and MMP-9 in keratinocytes obtained from OLP lesions exposed to hypoxia. The relative high expression of HIF-1α may be involved in multiple aspects of OLP progression through the regulation of its downstream target genes.

Blotting, Western ; Cell Hypoxia ; physiology ; Cell Proliferation ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Keratinocytes ; metabolism ; Lichen Planus, Oral ; pathology ; Matrix Metalloproteinase 9 ; metabolism ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; metabolism

Systematic research on the quality evaluation methods of Chinese medicinal materials is an intrinsic requirement,which is beneficial to the clinical application of traditional Chinese medicine and the sound development of traditional Chinese medicinal industry.The intrinsic quality evaluation methods of Chinese medicinal materials have developed from chemical fingerprint to quality marker of Chinese medicinal materials,and it represents that the evaluation mode has developed from chemical component as research index to the combination of chemical component and pharmacodynamic components,chemical component and biological activity.The extrinsic character evaluation method of Chinese medicinal materials from"Quality Evaluation Through Morphological Identification"to the application of intelligent sensory technology make up for the accumulation of objective data.The trend of quality evaluation of the intrinsic quality and extrinsic traits of Chinese herbal medicines conform to the innovation strategy called integrated quality control of TCMs.This paper reviews the development and evolution of the research on the intrinsic quality and extrinsic traits of Chinese herbal medicines,systematically expounds the research status and development trend of"intrinsic and extrinsic combination",and facilitate the establishment of quality evaluation system of Chinese herbal medicines with standardization and informatization characteristics in accordance with the theory of traditional Chinese medicine.

ObjectiveTo investigate the effects of spleen-Yin deficiency on gastrointestinal absorption， water metabolism and intestinal flora in rats with spleen-Yin deficiency syndrome. MethodA rat model of spleen-Yin deficiency syndrome was established by using the composite factors， including irregular meat and vegetable diet， weight-bearing fatigue swimming and gavage with warm-heat injury-Yin drugs. The changes of body weight， food intake， water intake and duration of swimming in the blank and model groups were observed. Hematoxylin-eosin（HE） staining was used to observe the histopathological damage of the stomach and colon. Urinary excretion rate of D-xylose was determined by phloroglucinol method. The content of gastrin（GAS） in serum was determined by enzyme-linked immunosorbent assay（ELISA）. The relative expression levels of vasoactive intestinal peptide（VIP）， aquaporin 3（AQP3） and AQP4 in gastric tissues were detected by Western blot. The relative mRNA expression levels of VIP， AQP3 and AQP4 in gastric tissues were detected by Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）， and the changes of intestinal flora were analyzed by 16S rDNA sequencing. ResultCompared with the blank group， the results of general physical signs showed that the body weight and food intake of rats in the model group were significantly decreased， the water intake was significantly increased（P<0.05， P<0.01）， and the duration of swimming was significantly decreased（P<0.01）. Pathological examination results showed that in the mucosa of gastric tissues of rats in the model group appeared to be misaligned， the mucosa of colonic tissues could be seen to be obviously thinned or mutilated， and the epithelial cells appeared to be necrotic or even exfoliated. Compared with the blank group， the urinary D-xylose excretion rate of rats in the model group was significantly decreased（P<0.01）， and the serum GAS content was significantly decreased（P<0.05）. Compared with the blank group， Western blot results showed that the relative expression level of VIP protein in gastric tissues of rats in the model group was significantly decreased， while the relative expression levels of AQP4 and AQP3 proteins were significantly increased（P<0.01）. Compared with the blank group， Real-time PCR results showed that the relative expression level of VIP mRNA in gastric tissues of rats in the model group was significantly decreased（P<0.01）， and the relative mRNA expression levels of AQP3 and AQP4 were significantly increased（P<0.05， P<0.01）. Compared with the blank group， the results of intestinal flora analysis showed that the number of operational taxonomic units（OTUs） and α-diversity increased and β-diversity decreased significantly in the model group， the abundance of Porphyromonadaceae was increased significantly， and the abundance of Oscillibacter_ruminantium was decreased significantly（P<0.05）. Spearman correlation analysis showed that Porphyromonadaceae was significantly positively correlated with AQP4 protein level， while Oscillibacter_ruminantium was significantly positively correlated with VIP protein level， and negatively correlated with AQP3 and AQP4 protein levels（P<0.05）. Linear discriminant analysis effect size（LEfSe） analysis results showed that there were significant differences in a variety of intestinal bacteria between groups， and the intestinal bacteria of the model group were significantly enriched in the phylum/order/family/genus of Elusimicrobia， Betaproteobacteria， Burkholderiales， Sutterellaceae and Parasutterella（P<0.05）. ConclusionSpleen-Yin deficiency syndrome can weaken the digestion and absorption capacity of gastrointestinal tract， and cause the disturbance of water metabolism and intestinal flora. AQP4， AQP3 and VIP protein levels of gastric mucosa are closely related to Porphyromonadaceae and Oscillibacter_ruminantium. And AQP4， AQP3 and VIP may be involved in the regulation of intestinal flora in order to affect the physiological function of spleen governing transportation and transformation.