Animals ; Apoptosis ; physiology ; Cell Proliferation ; Epithelial Cells ; transplantation ; Female ; Ki-67 Antigen ; analysis ; Kidney Tubular Necrosis, Acute ; pathology ; Male ; Mesenchymal Stem Cell Transplantation ; ethics ; Rats ; Rats, Sprague-Dawley

Background and purpose:Neuroendocrine neoplasm is one kind of infrequent neoplasms from neuroendocrine cell and peptidergic neurons. This study aimed to investigate the epidemiology and clinicopathological characteristics of neuroendocrine neoplasms (NEN) in Dalian. Methods: We retrospectively analyzed all neuroendocrine neoplasms patients in First Afifliated Hospital of Dalian Medical University from 2000 to 2013. The epidemiology characteristics, pathogenic sites and pathological types were reviewed, and the differences between gender and age were also analyzed. The statistics analysis such as independent-samples t test and one-way ANOVA and chi-square test were performed. Results:There were 279 NEN cases, including 166 male and 113 female patients (male:female=1.14:1), detection rate was 0.258%. The mean age of all cases was 59.4±17.1 (20-83), for male 58.9±19.4 (20-81) and female 61.7±15.0 (29-83). There was no statistical signiifcance of detection rate and mean age between male and female patients. Digestive system was the most common occurred site of NEN (71.68%), then was the respiratory system (20.79%). There was no statistical significance of pathogenic sites between male and female.In all case, 132 with neuroendocrine tumor (47.31%), 140 with neuroendocrine cancer (50.18%), 7 with mixed adenoneuroendocrine carcinoma (2.51%). There was no statistical signiifcance of pathological types between male and female (P>0.05).Conclusion:Recently, the detection rate of neuroendocrine neoplasms was raised in Dalian, but there was no signiifcance differences in detection rate,pathogenic sites and pathological types between male and female.

Objective To investigate the mechanism of traditional Chinese medicine (TCM) Sheshang capsule for treatment of blood coagulation dysfunction in patients bitten by Trimeresurus stejnegeri snake. Methods A prospective study was conducted. Seventy Trimeresurus stejnegeri snake envenoming patients whose manifestations conformed to the diagnostic criteria of the fire toxin syndrome in TCM were assigned into therapy group and control group by random number table (each, 35 cases). The basic treatments (including wound disinfection, intramuscular injection of 1 500 U tetanus antitoxin, conventional dose of antibiotics, 10 mg dexamethasone, 40 mg omeprazole) and 10 Jidesheng Sheyao tablets three times a day were applied in the control group. In the therapy group, the basic treatments the same as those of the control group were given, and in the mean time 5 Sheshang capsules (the drug was prepared in our hospital including ingredients:rhubarb, coptidis rhizoma, pleione bulbocodioides, elecampane inula root, bayberry bark, borneol and so on) were administered three times a day. The therapeutic course in the two groups was 1 week. The levels of platelet α-granule membrane protein (CD62p), thromboxane B2 (TXB2), platelet factor 3 (PF3) and von Willebrand factor (vWF) in serum were measured by enzyme linked immunosorbent assay (ELISA) before and after treatment. Results Before treatment, there were no significant differences in CD62p, TXB2, PF3 and vWF between therapy group and control group [CD62p (μg/L):3.81±1.64 vs. 3.52±1.43, TXB2 (μg/L):13.04±1.67 vs. 13.31±1.14, PF3 (μg/L): 2.84±1.08 vs. 2.88±1.23, vWF (μg/L):12.36±2.42 vs. 11.89±2.08, all P>0.05]. After treatment, the levels of CD62p, TXB2 and PF3 were increased, while vWF decreased compared with those before treatment in both groups, the level changes in therapy group being more remarkable [CD62p (μg/L): 6.73±1.77 vs. 5.81±1.62, TXB2 (μg/L):18.65±1.77 vs. 17.90±1.68, PF3 (μg/L):5.61±1.48 vs. 4.77±1.24, vWF (μg/L):3.87±1.01 vs. 4.58±1.09, P < 0.05 or P < 0.01]. Conclusion The Sheshang capsule is capable of treating patients with blood coagulative disorder after Trimeresurus stejnegeri snake bite, and its mechanism is possibly related to the improvement of platelet activation function and amelioration of the damage of vascular endothelial cells.

Site-saturation mutagenesis is a newly-developed technology in protein engineering.By manipulating the encoding genes,it can rapidly obtain the mutants of desired proteins whose target residues are substituted by 19 other common amino acids.Site-saturation mutagenesis could serve not only as a powerful tool in protein engineering,but also as an important method in exploring the structure-function relationship of proteins.Several techniques were summarized to achieve site-saturation mutagenesis and introduce their application status in the protein engineering.The problem and promising future of its application were also discussed.

Objective To investigate the influence of the purging fire and removing toxin method on chemokines and adhesion factors related to vascular endothelialitis injury induced by toxin of trimeresurus stejnegeri bite.Methods ① Animal experiment:50 healthy New Zealand white rabbits were chosen.According to random numbers generated by statistical software,they were divided into normal control group,model group,low,middle and high dose Sheshang capsule groups,10 in each group.Trimeresurus stejnegeri bite model was replicated by injecting 0.75 mL/kg snake venom into subcutaneous tissues of rabbits' right hind legs.And the same volume of normal saline was injected into the rabbit in the normal control group.After the model was established for 6 hours,the rabbits in low,middle and high dose Sheshang capsule groups received 174,348 and 522 mg· kg-1 · d-1 of Sheshang capsule solution respectively (the content of capsules was dissolved in normal saline to make liquid with 17.4,34.8 and 52.2 g/L Sheshang solution respectively,so the volume of gavage of each group was 10 mL· kg-1 · d-1);in the model and normal control groups,the same amount of normal saline was given by gavage,once daily for consecutive one week.24 hours after the last gavage,the blood of the rabbits was collected through an auricular border vein and the serum was separated by centrifuge ready for use.Meanwhile,the whole abdominal aorta segment of the rabbit was harvested and kept them in liquid nitrogen ready for use.② Cell experiment:human umbilical vascular endothelial cell (HUVEC) was cultured with MEM for 24 hours.The solution was replaced and according to the random number generated by statistical software,the cells were divided into blank control group,model group and low,middle,high dose Sheshang capsule medicinal serum groups,10 wells in each group.Trimeresurus stejnegeri toxin cell model was reproduced by addition of 5 mg/L snake venom into the cell culture medium.After 6-hour culture,the cells of model group and blank control group received 10％ normal rabbit serum,and the cells of low,middle and high dose Sheshang medicinal serum capsule groups received serum containing 5％,10％ and 15％ drug,respectively.After culture for 72 hours,the cells were collected and the total RNA was extracted.The real-time fluorescent quantitative polymerase chain reaction (qPCR) was used to detect the levels of mRNA of interleukin-8 (IL-8),monocyte chemoattractant protein-1 (MCP-1),intercellular adhesion molecule-1 (ICAM-1) and vascular endothelial cell adhesion molecule-1 (VCAM-1) in the vascular endothelial cells of rabbit aorta abdominalis and human umbilical vein,and the content of serum E-select element (CD62E) was measured by enzyme linked immunosorbent assay (ELISA).Results In model group,the expression levels of mRNA in IL-8,MCP-1,ICAM-1,VCAM-1 and the content of CD62E were all increased significantly in the endothelial cells of rabbit aorta abdominalis and HUVEC compared with those in control group [when the mRNA expression levels of IL-8,MCP-1,ICAM-1 and VCAM-1 in normal and blank control group were all being 1,the mRNA expression levels (2-△ △Ct) of the above mentioned inflammatory factors and adhesion molecule in animal model group were 3.96 ± 0.39,3.07 ± 0.27,3.71 ± 0.26,3.94 ± 0.26,and the mRNA expression levels (2-△ △Ct) of the above mentioned inflammatory factors and adhesion molecule in HUVEC model group were 3.53±0.70,2.24±0.48,3.13±0.44,2.80±0.13,respectively,all P ＜ 0.01].The content of CD62E in serum was increased significantly in model group compared with that in normal control group (μg/L:1.31 ± 0.22 vs.0.82 ± 0.13,P ＜ 0.01),the mRNA expression levels of IL-8,MCP-1,ICAM-1 and VCAM-1 were decreased significantly in low,middle,high dose Sheshang capsule groups compared with those in model group in endothelial cells of aorta abdominalis of rabbits and HUVEC [abdominal aorta:IL-8 mRNA (2-△ △Ct) were 1.13 ± 0.19,1.26 ± 0.16,1.27 ± 0.17 vs.3.96 ± 0.39,MCP-1 mRNA (2-△ △ Ct) were 1.79 ± 0.24,2.22 ± 0.38,1.76±0.19 vs.3.07±0.27,ICAM-1 mRNA (2 △△Ct) were 2.05±0.11,1.68±0.09,2.37±0.48 vs.3.71±0.26,VCAM-1 mRNA (2-△△Ct) were 1.59±0.08,1.40±0.11,1.84±0.11 vs.3.94±0.26;HUVEC:IL-8 mRNA (2-△△Ct) were 2.33±0.59,2.82±0.82,2.51±0.77 vs.3.53±0.70,MCP-1 mRNA (2-△△Ct) were 1.59±0.35,1.48±0.36,1.54±0.29 vs.2.24±0.48,ICAM-1 mRNA (2-△△Ct) were 1.46±0.38,1.77±0.65,1.73±0.50 vs.3.13±0.44,VCAM-1 mRNA (2-△△Ct) were 2.49±0.24,2.18±0.19,2.45±0.24 vs.2.80±0.13,all P ＜ 0.05].The contents of CD62E were decreased significantly in middle,high dose Sheshang capsule groups compared with the content in model group (μg/L:1.01 ±0.14,1.04±0.13 vs.1.31 ±0.22,all P ＜ 0.01),but there were no statistical significant differences among the three drug group (all P ＞ 0.05).Conclusion The therapy of purging fire and removing toxin can treat vascular endothelial injury by inhibiting the inflammatory response induced by Trimeresurus stejnegeri bites.

To investigate the role of volatile components in the compound and to find the substance foundation of Gualou Guizhi decoction (GLGZD) for curing extremities spasticity after stroke. The chemical compositions of essential oil, obtained by hydrodistillation from Gualou Guizhi decoction and its major constituting herbs (Trichosanthis Radix, Paeoniae Alba Radix, Cinnamomi Ramulus, Zingiberis Recens Rhizoma, Glycyrrhizae Radix, Ziziphi Jujubae Fructus) were analyzed by GC-MS to evaluate the correlativity between volatile components of GLGZD and its major constituting herbs, and volatile components after oral administration of GLGZD in the rats' brain. Volatile components of GLGZD are mainly derived from Cinnamomi Ramulus, Zingiberis Recens Rhizoma, Ziziphi Jujubae Fructus, Trichosanthis Radix. The volatile components in the brain is mostly derived from radix trichosanthis. Compared with individual herbs of GLGZD, the dissolution of the components increase or new components appear after compatibility of six herbs. Adminstrated with GLGZD, the results point out that volatile components in the brain play a neuroprotective role through passing the brain.
Animals ; Brain ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Gas Chromatography-Mass Spectrometry ; Male ; Rats ; Rats, Sprague-Dawley ; Volatile Organic Compounds ; chemistry ; pharmacology

This study is to determine six chlorogenic acids (chlorogenic acid, 5-O-caffeoylquinic acid, 4-O-caffeoylquinic acid, 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid and 4,5-di-O-caffeoylquinic acid) by quantitative analysis of multi-components with a single marker (QAMS). Chlorogeinc acid was used as internal reference to calculate the relative correction factors (RCF) of five compounds. Then the ruggedness of relative correction factors was tested on different instruments and columns. Meanwhile, a total of 4 batches of Lonicerae Flos and 20 batches of Lonicerae Flos extract with five different processing procedures were analyzed by external standard method (ESM) and QAMS, respectively. The ruggedness of relative correction factors was good. And the analytical results calculated by ESM and QAMS showed no difference. The quantitative method established was suitable for the quality evaluation of Lonicerae Flos extract.
Biomarkers ; analysis ; Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Flowers ; chemistry ; Lonicera ; chemistry

Animals ; Cell Culture Techniques ; methods ; Cell Separation ; methods ; Culture Media ; Epithelial Cells ; cytology ; Respiratory System ; cytology ; Swine