Objective To evaluate the effect of α-melanocyte stimulating hormone (α-MSH) and its novel analogue STY39 on the production of tissue factor pathway inhibitor (TFPI) in mice with endotoxemia.Methods Female BALB/c mice were randomly divided into eight groups with 9 mice in each group.Endotoxemia was reproduced by intraperitoneal injection of lipopolysaccharide (LPS,25 μg/kg) and D-galactosamine (D-Gal,100 mg/kg).The animals of the control group were given phosphate buffered solution (PBS) instead.In the experimental groups,the mice were injected intraperitoneally with 2.5 mg/kg α-MSH or STY39 at 1,2 or 3 hours following LPS injection.The orbital blood was collected at different time points,and tissues of lung,liver,and kidney were collected 8 hours after the administration of LPS.The plasma TFPI levels were determined by enzyme linked immunosorbent assay (ELISA),and the expression of TFPI mRNA in different tissues was determined with reverse transcription-polymerase chain reaction (RT-PCR).Results The plasma TFPI levels (μg/L) began to increase (11.84 ± 1.55) in the endotoxemia mice 4 hours after LPS challenge and reached the peak (23.49 ± 1.12) at 8 hours.α-MSH or STY39 treatment at 1,2 or 3 hours after LPS challenge could significantly increase the TFPI content,with the best drug effect at 1 hour after LPS challenge (the blood was collected 8 hours after LPS challenge,α-MSH group:58.79 ± 2.67 vs.28.49 ± 1.69,STY39 group:71.08 ± 2.13 vs.28.49 ± 1.69,both P＜0.01),and the effect of STY39 was better than that of α-MSH (P＜0.01).A small amount of TFPI mRNA expression was observed in each tissue of the healthy mice.After LPS challenge,TFPI mRNA expression was increased in all the tissues,especially in the lung,liver and kidney.α-MSH or STY39treatment at 1 hour after LPS challenge could significantly up-regulate the expression of TFPI mRNA in the lung and liver (A value,α-MSH in lung:51.10 ±2.89 vs.32.43 ±2.51,STY39 in lung:72.11 ±3.48 vs.32.43 ±2.51;α-MSH in liver:43.21 ± 2.12 vs.29.29 ± 2.06,STY39 in liver:66.82 ± 1.76 vs.29.29 ± 2.06,both P＜0.01).The treatment with STY39 at 1 hour after LPS challenge could significantly up-regulate the expression of TFPI mRNA in the kidney (A value:45.21 ± 1.80 vs.30.44 ± 2.23,P＜0.01),but the treatment with α-MSH had no obvious effect (A value:24.61 ± 1.98 vs.30.44 ± 2.23,P＞0.05).The enhancing effect of early administration of STY39 on TTPI mRNA expression in the lung,liver and kidney tissues of endotoxemia mice was more powerful than that of α-MSH (all P＜0.01).Conclusion The early administration of α-MSH or STY39 may up-regulate TFPI production in the mice with endotoxemia,and the effect of STY39 is superior to α-MSH.

Objective To observe the serum inflammatory factors level in patients with obstructive sleep apnea syndrome (OSAS) and coronary heart disease (CHD).Methods Venous blood was collected at 6 AM from 53 natiems with both OSAS and CHD and 37 simple snorers following a full night polysomnography (PSG) test.Serum MMP-9、TIMP-1 test were conducted.Results MMP-9,TIMP-1 level in patients with OSAS and CHD was significantiv higher than that of the simple snorers (P<0.05).Conclusion serum MMP-9 and TIMP-1 level increase in patients with OSAS and CHD.It is suggested that elevated level of serum MMP-9 and TIMP-1 might play an important role in the development of CHD in patients with OSAS.

Objective To study the effect of alpha-melanocyte stimulating hormone (α-MSH) and its novel analogue ( STY39 ) on the production of tissue factor ( TF ) and tissue factor pathway inhibitor (TFPI) stimulated by lipopolysaccharide (LPS) in primary mouse brain microvascular endothelial cells (MBMECs). Methods Female BALB/c mice were selected,purified and primarily cultured for 5 to 7 days. Immunofluorescence assay was use to detect the Ⅷ factor related antigen and identify the MBMEC model. The MBMECs were divided into eight groups:PBS control group, LPS stimulation group, after LPS stimulation 1,2,and 3 h adding 10 -7 mol/Lα-MSH groups or STY39 group (LPS+α-MSH,LPS+STY39) ( n=4 wholes in each group) . The cell culture supernatant and cells were collected at 6 and 8 h after LPS stimulation. An enzyme-linked immunosorbent assay was used to detect the concentrations of TF and TFPI in cell supernatant. RT-PCR was used to detect the expression levels of TF mRNA. Results (1) LPS could induce MBMEC to produce TF and TFPI proteins. The level of TF in the cell culture supernatant reached the peak at 6 h,and the level of TFPI reached the peak at 8 h. (2) At 1,2,and 3 h after LPS stimulating MBMEC,10 -7mol/L α-MSH or STY39 were given. They could significantly decrease the TF protein content in the cell supernatant (P<0. 01),especially the effects of giving α-MSH or STY39 were most significant at 1 h after LPS stimulation (P<0. 05). The effect of STY39 for decreasing TF content was more significant than that of α-MSH (P<0. 05);however,α-MSH and STY39 did not have significant up-regulating effects for LPS inducing MBMEC to produce TFPI. (3) After LPS stimulation,10 -7 mol/Lα-MSH or STY39 were given at different time points. They significantly down-regulated the expression level of MBMEC TF mRNA (P<0. 01). The effect was most significant at 1 h time point (P<0. 05),but there was no significant difference in the effects betweenα-MSH and STY39. Conclusion Bothα-MSH and STY39 can suppress LPS-induced primary MBMEC to produce TF protein and express TF mRNA,and the effect of administration is better after 1 h LPS stimulation. The suppressive effect of STY39 on the production of TF protein is superior toα-MSH.

Objective To study the morphology changes and the expression of phosphatidyl serine(PS) in apheresis platelets(Plts) during storage,also to analyze their relationship,in order to provide laboratorial evidence for the in vitro storage period of plts.Methods The morphological changes of plts during 8-day storage were detected by May-Grunwald-Giemsa dyeing method,and the expression of PS on platelet membrane was tested by flow cytometry(FCM).Results The morphological lesions appeared on the fourth day of storage,because the modified morphological score(MMS) of 4-day group was significantly lower than that of fresh plts(t=2.341,P

Objective:To investigate therapeutic mechanism of immunoglobulin Yolk (IgY) against tumour necrosis factor alpha ( TNF-α) and interleukin-1 beta ( IL-1β) in guinea pigs with allergic rhinitis.Methods: Hartley guinea pigs were randomly divided into the control group (group C,n=17),the allergic rhinitis model group (group M,n=27),the 0.1%anti-TNF-αand IL-1βIgY treating group (group Z1,n=21) and the fluticasone propionate treating group (group Z2,n=21).The allergic rhinitis model in guinea pigs was established using ovalbumin.After treatment for 2 h,4 h,8 h,nose and bronchial lung were lavaged using 0.9%saline, the nasal lavage fluid (NLF) and bronchoalveolar lavage fluid (BALF) were collected,the precipitated cells were stained using Wright′s,the nasal mucosa and lung tissues were stained using methylene blue and eosin (HE),and TNF-α,IL-1β,IL-5 and IL-33 in nasal mucosa and lung tissues were stained using immunohistochemistry.Results:There were a large amount of eosinophils and more serious inflammation responses in nasal mucosa in the M group compared with the Z 1 and Z2 groups.In the lung tissues,there were more alveolar tube damage ,pulmonary interstitial edema ,interval thickening ,thickening of bronchial smooth muscle and inflammation cell in-filtration in the M group compared with the Z 1 and Z2 groups.The eosinophils ,lymphocytes and neutrophils were significantly decreased in NLF and BALF in the Z1 and Z2 groups compared with the M group (P<0.05).The expressions of IL-1βand TNF-αfrom 2 h to 8 h and IL-5 and IL-33 from 4 h to 8 h significantly decreased in the nasal mucosa and lung tissues in the Z 1 group compared with the M group ( P<0.05 ).Conclusion:The allergic rhinitis in guinea pigs accompany with the allergic asthma.The inhibitory capacity of anti-TNF-αand IL-1βIgY on pathological responses in guinea pigs with allergic rhinitis may be due to the significant decrease in the infiltration of eosinophils and the expressions of inflammatory cytokines in the nasal mucosas and lung tissues .

Drugs, Chinese Herbal ; pharmacology ; Heart ; Humans ; Liver ; Organ Preservation Solutions ; Panax ; Pancreas ; Salvia miltiorrhiza

Objective To analyze the features of EMU survived casualties and the rescue during a head -on- rear collision between two EMU trains on 23 July 2011 ( July 23 train collision accident) at Wenzhou station.Methods The casualties treated in many major hospitals in Wenzhou were surveyed within 24 hours after the accident occurred.The data of age,gender,type of injury and injury severity of the wounded were analyzed.Results A total of 136 casualties were treated within the first 24 hours after the accident occurred,and the male patients and female patients accounted for 55.89％ and 44.11％ respectively,blunt trauma was the main cause of injuries.The percent of multiple injuries in the wounded survivals accounted for 79.41％.The most common injury site of the survived casualties was chest,followed by four limbs and spine.All the wounded were rescued on the spot and were referred to the hospitals with better medical facilities.Conclusions There was no significant difference in gender of the wounded.Blunt trauma was the leading cause of injuries,and the chest,four limbs and spine were the liable parts of body to be traumatized.Saving life,triaging and transferring the wounded as soon as possible were the major algorithm during the initial stage of medical rescue after the accident occurred.

Objective\ To observe the expression of 5\|HT 1A and 5\|HT 2A receptor mRNAs in the rat spinal cord and dorsal root ganglion(DRG). Methods\ In situ hybridization histochemical technique. Results\ (1)5\|HT\-\{1A\} receptor mRNA positive neurons were found in all laminae of the gray matter,mainly in the superficial laminae(laminae Ⅰ and Ⅱ),laminae Ⅲ and Ⅳ of the dorsal horn.Scattered positive neurons were also observed in laminae Ⅴ\|Ⅶ and Ⅹ.Very few positive signals were found in the ventral horn(lamnina Ⅸ);(2)5\|HT\-\{2A\} receptor mRNA positive neurons were mainly found in the superficial laminae and ventral horn,while sparsely distributed positive neurons were also located in other laminae.Within the DRG:(1)About 10\^4% of the total DRG cells were labeled with 5\|HT 1A receptor mRNA.The positive signals were mainly confined to a subpopulation of small\| and medium\|sized cells;(2)About 17\^4% of the total DRG cells were labeled with 5\|HT 2A receptor mRNA,most of them were also small\| and medium\|sized cells. Conclusions\ 5\|HT 1A and 5\|HT 2A receptor mRNAs positive neurons distributed heterogeneously in the rat spinal cord and DRG, they may play important roles for 5\|HT in the analgesic effects at the spinal level and nociception in the periphery.\;

Background JAK/ signal transducer and activator of transcription 3 (STAT3) signal pathway plays a critical role during the sclera remodeling of experimental myopia.As a tyrosine kinase inhibitor,AG490 can inhibit the activation of this pathway.But whether AG490 plays a role in delaying the development of myopia is not completely clear.Objective This study was to investigate the inhibition of AG490 to activation of STAT3 signaling pathway and the sequential arresting effect on the sclera remodeling in form-deprived myopia (FDM) models.Methods Forty guinea pigs were randomly divided into the normal control group,model control group,PBS control group and AG490 treatment group.FDM models were established by the occlusion of the right eyes of guinea pigs for consecutive 4 weeks using translucent goggles in the model control group,PBS control group and AG490 treated group,and 25 μl PBS or AG490 were respectively injected into vitreous since the first day of modeling in two-day interval till the fourth week in the PBS control group and AG490 treated group.Refractive state and axial length were examined with retinoscopy and A-scan ultrasonography before and 4 weeks after experiment.The experimental eyes were extracted in the fourth week,and the expressions of scleral STAT3,p-STAT3,metal matrix proteinase-2 (MMP-2) proteins and STAT3 mRNA,MMP-2 mRNA were detected by immunocytochemstry and semi-quantitative reverse transcription PCR (RT-PCR) respectively.The use and care of experimental animals followed ARVO.Results Compared to the normal control group,the negative refraction power and axial length were significantly increased in the model control group,PBS control group and AG490 treated group,and the axial length in the AG490 treated group was smaller than those in the model control group and PBS control group,showing significant differences among the 4 groups (refraction:F =89.063,P =0.000;axial length:F =96.145,P =0.000).The expressions of STAT3,MMP-2 and p-STAT3 in scleral tissue were weaker in the normal control group.The expressional values (A values) of STAT3,p-STAT3 and M MP-2 were 0.064 ± 0.016,0.019 ± 0.002 and 0.155 ± 0.052 in the AG490 treated group,which were lower than 0.129±0.008,0.071 ±0.021,0.425 ±0.004 of the model control group and 0.130±0.004,0.069±0.002,0.421 ±0.042 of the PBS control group (STAT3:t =4.641,9.364,both at P＜0.01;p-STAT3:t =4.638,4.488,both at P＜ 0.05;MMP-2:t =9.123,9.029,both at P ＜ 0.05),however,these expressions were still higher than those of the normal control group (t =2.674,2.251,2.682,all at P ＜0.05).The expressional levels (A values) of STAT3 mRNA and MMP-2 mRNA in the AG490 treated group were 0.295±0.032 and 0.569±0.019,which were significantly lower than 0.547±0.015 and 0.782±0.051 in the model group as well as 0.544±0.015 and 0.779±0.048 in the PBS control group (STAT3 mRNA:t =10.115,11.703,both at P＜0.01;MMP-2 mRNA:t =9.218,9.494,both at P＜0.01).The expressional levels (A values) of STAT3 mRNA and MMP-2 mRNA in the AG490 treated group were still higher than those in the normal control group (t=2.576,3.565,both at P＜0.05).Conclusions AG490 can ultimately inhibit the development of axial myopia by arresting the activation of STAT3 signaling pathway in the FDM eyes and further regulating the expression of MMP-2 in sclera and delaying the remodeling of sclera.

Objective:To identify metastasis-associated genes in salivary gland adenoid cystic carcinoma(ACC).Methods:Salivary gland adenoid cytic carcinoma cell line ACC-2 and its highly metastatic ACC-M cells were used to screen the metastasis-related genes in ACC by microarray technology.Two fluorescent cDNA probes labeled with Cy3 and Cy5 dyes,were prepared from the mRNA samples of ACC-2 and ACC-M cells by reverse transcription method.The two color probes were then mixed and hybridized on the cDNA chip constructed by double dots of 1152 human genes,and scanned at two wave lengths.Differentialy expressed genes of the two cell lines were analyzed using computer.Then seven of the differently expressed genes were further validated by RT-PCR technique.Results:Of the 1,152 known genes and expressed sequence tags,26 showed significantly different expression level(minimum 2 fold) between the two cell lines.Among the 26 genes,19 were up regulated(with ratio more than 2) and 7 were down(with ratio less than 1/2).The results of RT-PCR analysis for 7 differently expressed genes were coincident with those of microarray assay.Conclusions:Down regulation of LIFR,LCP1,DPEP1 and ABLIM1,and up regulation of DCC,MMP1 and CNTN2 may be related to the highly metastatic potential of ACC-M cell line.