To explore the common irritant components in different base sources of Polygonati Rhizoma(PR). A rabbit eye irritation experiment was conducted to compare the irritant effects of raw products of Polygonatum kingianum, P. officinale, and P. multiflorum. The irritant effects of different solvent extraction parts and needle crystals of PR were compared, and the irritant components were screened. The morphology and structure of the purified needle crystal of PR were observed by microscope and scanning electron microscope and characterized by X-ray diffraction. Rabbit eye irritation and mouse abdominal inflammation model were used to evaluate rabbit eye irritation scores, inflammatory mediators, inflammatory factors levels in the peritoneal exudate of mice, with the peritoneal pathological section used as indicators. The inflammatory effect of needle crystals of PR was studied, and the content of calcium oxalate in three kinds of PR was determined by HPLC. The common protein in three kinds of PR was screened and compared by double enzymatic hydrolysis in solution combined with mass spectrometry. The results showed that three kinds of PR raw products had certain irritant effects on rabbit eyes, among which P. kingianum had the strongest irritant effect. There were no obvious irritant effects in the different solvent extraction parts of P. kingianum. Compared with the blank group, the needle crystal of PR had a significant irritant effect on rabbit eyes, and the inflammatory mediators and inflammatory factors in the peritoneal exudate were significantly increased(P<0.05) in a dose-dependent manner. Meanwhile, the peritoneal tissue of mice was damaged with significant inflammatory cell infiltration after intraperitoneal injection of needle crystal, indicating that needle crystal had an inflammatory effect. Microscope and scanning electron microscope observations showed that the needle crystals of PR were slender, with a length of about 100-200 μm and sharp ends. X-ray diffraction analysis showed that the needle crystals of PR were calcium oxalate monohydrate crystals. The results of HPLC showed that the content of calcium oxalate in P. kingianum was the highest among the three kinds of PR. It was speculated that the content of needle crystal in P. kingianum was higher than that in P. officinale and P. multiflorum, which was consistent with the results of the rabbit eye irritation experiment. The results of mass spectrometry showed that ribosome inactivating protein and mannose/sialic acid binding lectin were related to inflammation and cell metabolism in all three kinds of PR. There was no obvious irritant effect in different solvent extracts of PR. The calcium oxalate needle crystal contained was the main irritant component of PR, and three kinds of PR contained common ribosome inactivating protein and mannose/sialic acid binding lectin, which may be related to the inflammatory irritant effect of PR.
Animals ; Rabbits ; Mice ; Polygonatum/chemistry* ; Drugs, Chinese Herbal/toxicity* ; Rhizome/chemistry* ; Male ; Eye/drug effects* ; Female ; Humans

ObjectiveThe pathophysiological changes caused by trauma are complex, and there is still a lack of effective markers for injury monitoring and prognosis judgment. This paper aims to investigate the relationship between neutrophil/lymphocyte ratio (NLR) and lactate level and trauma severity and prognosis in trauma patients.MethodsThe data of 169 trauma patients who met the inclusion criteria in the Department of Emergency Medicine, General Hospital of the Eastern Theater Command from January to December 2017 were retrospectively analyzed. Their gender, age, cause of injury, injury severity score (ISS), acute physiology and chronic health evaluation II (APACHE II), Glasgow coma index (GCS), NLR and lactate levels within 24 hours after the injury and other laboratory results were recorded. According to the ISS score, the patients were divided into mild injury group (ISS<16, n=78), severe injury group (16≤ISS<25, n=53), and critical injury group (ISS≥25, n=38); According to the 28-day outcomes, they were divided into death group (n=22) and survival group (n=147). The correlation between NLR and lactate levels and ISS score was analyzed, and the prognostic value was analyzed by receiver operating characteristic (ROC) curve.ResultsNLR was positively correlated with ISS (r=0.34, P<0.001), and there were significant differences among three trauma groups (P<0.001). Lactate was positively correlated with ISS (r=0.28, P=0.002) too, and significant differences were shown among three trauma groups (P=0.003). The area under the ROC curve of NLR and lactate for predicting 28-day death in trauma patients was 0.785 and 0.686, respectively.ConclusionNLR and lactate are positively correlated with trauma severity and they might play an important part in the early prognosis evaluation of trauma patients.

OBJECTIVETo construct TIMP-1 siRNA eukaryotic expression vectors and evaluate their effect on TIMP-1 mRNA expression in hepatic stellate cells.

METHODSThe combinant lone DNA with cutting sites of BamH I and Xho I enzyme according to the sequences of 447-465, 552-540 TIMP-1 of rats and nonspecific sequence were selected and cloned to pGEM-T vector and sub-cloned to pRNAT-U6.2. They were then identified by double enzyme digestion analysis and DNA sequencing. Three plasmids were transfected into T6 separately through an oligofectamine package. TIMP-1 mRNA expression was evaluated by RT-PCR.

RESULTSTargeting sequences of TIMP-1 siRNA eukaryotic expression vectors were correct. TIMP-1 mRNA expression was significantly reduced by transfecting them into the T6.

CONCLUSIONWe successfully constructed two TIMP-1 siRNA eukaryotic expression vectors and the transfected cells can significantly suppress the TIMP-1 expression.

Animals ; Cell Line ; Gene Silencing ; Genetic Vectors ; Hepatic Stellate Cells ; Plasmids ; RNA, Small Interfering ; Rats ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; Transfection