Objective To investigate the influence of seawater immersion of open abdominal wound on function and pathological changes in liver in dog.Methods Twenty healthy dogs were divided randomly into two groups.A perforating abdominal wound was was made in each animal of both groups.Animals in control group(n=10)only abdominal wound was made,while abdominal wound was followed by seawater immersion in the animals of seawater immersion group(n=10).Blood samples were taken before the trauma(0 hour)and at 4,8,12,16 and 20 hours after the trauma and water immersion for both groups.Total bilirubin(TB),alanine transaminase(ALT),aspartate transaminase(AST),lactate dehydrogenase(LDH),blood ammonia(NH3),prothrombin time(PT),tumor necrosis factor-?(TNF-?),interleukin-6(IL-6)and endotoxin(ETX)were assayed.Experimental animals were sacrificed 24 hours after trauma,and the pathological changes in hepatic tissue were studied.Results TB,ALT,AST and PT were higher in seawater immersion group,than those in control group,with a marked elevation at 16 hours.In seawater immersion group,LDH was significantly higher than those in control group at 4 and 12 hours,and NH3 was increased significantly at 4 hours in experimental group(P

Autoimmune Diseases ; chemically induced ; Female ; Humans ; Interferons ; adverse effects ; Purpura, Thrombocytopenic, Idiopathic ; chemically induced ; Young Adult

objective: To study the effects of BMP on the growth of osteosarcoma cells. Methods: Human antisense BMP2 retrovirus expression vector was constructed and transfected into human osteosarcoma OS 9901 cells by Lipofect AMINE. Positive cell clones were selected with G 418. The expression of BMP and PCNA were determined by immunohistochemical methods, image analysis and expressed as grey level. The morphology and cell cycle distribution of the cells were studied by electronmicroscope and flowcytometry respectively. Results:The grey levels of BMP and PCNA in the transfected cells were 198.4?8.51 and 197.3?3.22, those of the control were 135.1?12.32 and 142.9?8.47,respectively. G 1, G 2 and S phase cells were 51.9%,18.2% and 22.7% in the cell cycle of transfected cells, while those of the control were 52.8%,11.1% and 36.1%, respectively.Increased lysosome and small pieces of chromatin were observed in the transfected cells under electronmicroscope. Conclusion:Transfection of antisense BMP2 may inhibite the expression of BMP and proliferating activity of osteosarcoma cells.

Objective To establish an AGS cell line that stably expressing miR?126 and to study the effect of miR?126 on the proliferation and metastatic abilities of the AGS cell line in vitro. Methods AGS cells were infected by lentivirus with Lv?has?mir?126. After confirmation by RT?PCR ,CCK?8 and clone formation assays were used to evaluate the effect of miR?126 on AGS cell growth. Transwell migration and invasion assays were used to evaluate the effect of miR?126 on metastasis of AGS cells. Results We verified correct construction of recombinant AGS cells. RT?PCR confirmed mRNA levels of miR?126 existed significantly differences among the recombinant cell lines (P< 0.05). Proliferation assays and clone formation assays did not show a remarkable growth suppression in AGS?mir?126 cell line. However,transwell assay showed a notable acceleration in AGS?mir?126(P< 0.05). Conclusions We successfully constructed recombinant AGS cell line with stably high miR?126 expression level. MiR?126 could facilitate the metastasis of AGS cell in vitro.

AIM:To investigate the protective effect of nerve growth factor combined with Ginkgo biloba extract on retinal ischemia-reperfusion (RIR) injury in rabbits with experimental high intraocular pressure.METHODS:Establishment of rabbit glaucoma ischemia reperfusion model.Twenty-four New Zealand white rabbits were randomly divided into three groups:nerve growth factor group, Ginkgo biloba extract group and combination group.Respectively, in the continuous administration of 1, 7, 14d.We observed the morphological changes of the tissues of the retina.The levels of superoxide dismutase(SOD), nitric oxide(NO) and malondialdehyde(MDA) in retinal tissue were measured.RESULTS:Respectively, first, in the continuous administration of 1, 7, 14d, the contents of MDA and NO in Ginkgo biloba extract group and nerve growth group were higher than that in combination group (P<0.05).Secondly, the SOD content of Ginkgo biloba extract group and nerve growth group were lower than that of combination group at each time point (P<0.05).At each time point, the number of HE staining of retinal ganglion cells (RGCs) showed that the loss of RGCs in the combination group was significantly lower than that in the other groups, and the ganglion cell count showed that the Ginkgo biloba extract group and the neuronal growth group were lower (P<0.05).CONCLUSION:Nerve growth factor combined with Ginkgo biloba extract has better protective effect on retinal ischemia-reperfusion injury.The mechanism may be related to the decrease of free radicals and increase the activity of SOD in retinal tissue.

Objective To evaluate the biochemical function of rat nature three-dimensional acellular pancreatic bioscaffold (3D-APB) in promoting cell proliferation and differentiation in vitro.Methods The fresh pancreas from 10 rats were perfused to prepare 3D-APB.The biocompatibility of 3D-APB was eva luated.The experiment was divided into 4 groups based on 4 kinds of three-dimensional media for AR42J culture,including blank control group,ECM group,PLGA group and 3D-APB group.We compared the proliferation and differentiation of AR42J pancreatic acinar cell at 3 days,5 days,7 days and 10 days after seeding among 4 groups.Results The 3D-APB could represent a biocompatible scaffold capable of integrating within host tissue.The proliferation rate of AR42J cells by MTT in 3D-APB was higher,while the apoptosis rate by flow cytometry was lower than those in other 3 media,which were all significantly different (P ＜ 0.05),respectively.The protein expression of PDX-1 and PTF-1 by western blot in 3D-APB group was greatly higher than those in other 3 groups (both P ＜ 0.05).The mRNA expression of PDX-1 and PTF-1 through qRT-PCR was significantly higher than those in other 3 groups (both P ＜ 0.05).Conclusion Compared with the commonly used chemical and natural scaffold at present,3D-APB could promote cell proliferation and differentiation,which was more appropriate for regenerative medicine.

Objective To explore the influences of age and duration of status convulsion (SC) on hippocampal neuron apoptosis by observing the dynamic change of neuron apoptosis in rats with different age when SC terminates. Methods Seizures were induced in infant rats (IRs) and adult rats (ARs) injected with lithium and pilocarpine intraperitoneally. Rats were sacrificed at 6 time points (3, 6, 12 hours and 1, 3, 7 days) after 30 minutes of SC, or 2 time points (3 hours and 1 day ) after 3 hours of SC respectively. The location and type of apoptotic cells were assessed by using terminal deoxynucleotidyl dUTP nick end-labeling (TUNEL) of brain sections in situ. The proportion of apoptotic cells was quantified by Annexin-Ⅴ-FITC apoptosis detecting method and analysed by flow cytometer. Results (1) SC induced neuronal apoptosis and necrosis mainly in the CA_1 and CA_3 regions of hippocampus. (2) As compared to the time point before SC, the proportion of apoptotic cells in IRs and ARs hippocampus was increased obviously at 3 hours point after 30 minutes of SC (IRs 0.55%?0.21%, ARs 0.53%?0.06%), and with a maximal induction at 12 hours in IRs (0.67%?0.18%) and 1 day in ARs (0.98%?0.38%). The apoptotic process continued at least for 3—7 days. (3) In IRs, the proportion of apoptotic cells was lower than in ARs at different time points after 30 minutes of SC, except 3 hours point. There was a significant difference between the two age groups at 1 day and 7 days after SC respectively. The age-related difference was more obvious after 3 hours SC. Conclusions (1) Severe seizure induces neuronal apoptosis in hippocampus. (2) Age and duration of SC might be the important factors in influencing the neuronal apoptosis. The neuronal apoptosis in hippocampus after SC would have a positive correlation with age and duration of SC.

Objective To evaluate neurodevelopmental outcome of very preterm(gestational age ＜ 32 weeks) and very low birth weight infant (VLBWI) (weight ＜ 1 500 g) and to examine the effectiveness of an early intervention program till 12 months corrected age.Methods Seventy followed-up very preterms and VLB-WI in Jinan Maternity and Childcare Hospital were enrolled in this study from January 2012 to and were divided into two groups by birth weight.All infants received 20 items of behavioral neurological assessment at 1 to 12 months corrected age and tested mental and psychomotor development with the use of CDCC at 6,12 months corrected age.The preterms who were abnomal in the 20 items of behavioral neurological assessment would receive early intervention (including kinesitherapy, physiotherapy, cereal circulation therapeutic equipment) by physiotherapists and their parents who received an intervention program training and were strongly encouraged to participate in the intervention sessions.The intervention method was adjusted according to the neurological assessment.The SPSS statistical software package for Windows, version 15.0, was used to run Fisher's exact test and t-test on the data presented,and P value of less than 0.05 was regarded as statistically significant.Results The average gestational age of infants was (30.4 ± 1.8) weeks,and average birth weight (1 463.7 ± 307.5) g.The incidence of extrauterine growth restriction was 57.1％ at first follow-up.The incidence of neurodevelpmental impairment NDI) and cerebral palsy tendency at 6 corrected months were 14.3％ ,8.6％ respectively.At 12 months corrected age,the incidence of NDI decreased to 2.9％ and cerebral palsy to 2.9％.There was significant difference in the incidence of NDI between 6 and 12 corrected months.There was no significant difference in the incidence of psychomotor developmental index ＜ 70, mental developmental index ＜ 70, NDI and cerebral palsy between the two groups.Conclusion The early intervention program can improve VLBWI neurodevelopmental outcomes at 12 months' corrected age and reduce the incidence of cerebral palsy.

The correct pairing of disulfide bonds maintains the correct folding mode and high-level structure formation of peptides and protein drugs, which is crucial for the quality control of products. In order to ensure that the disulfide bonds are correctly paired, disulfide bond analysis is an essential part of peptides and protein drug characterization. Mass spectrometry can be used to analyze disulfide bonds. However, insulin and its analogues have two pairs of disulfide bonds without restriction enzyme cutting site. Conventional collision-induced dissociation (CID) and high-energy induced cleavage (HCD) cannot accurately locate the complex disulfide bond. In our study, three methods were used to localize the complex disulfide, including enzyme digestion combined with key peptide fragment in source decay (ISD) fragmentation method, enzyme digestion combined with partial reduction alkylation method, intact protein source ISD and electron transfer dissociation (ETD) cleavage method, The applicability of insulin aspart, insulin lispro and insulin glargine were also investigated. This study provides a new way for the quality control of disulfide bonding mode of insulin and its analogues, and also provides a reference for the disulfide bond localization of peptides or proteins containing this complex disulfide bond.

Objective To investigate the effects of using a catheter balloon to treat crieopharyngeal achala-sia in patients with brainstem lesions. Methods Thirty cases of dysphagia caused by brainstem lesions were diag-nosed as crieopharyngeal achalasia through videofluoroscopy of swallowing.The cases were divided into a treatment group and a control group randomly.The treatment group was treated with balloon dilatation and routine dysphagia re-habilitation training once daily,while the control group was treated with routine dysphagia rehabilitation training only.The treatment end point was either the patient resuming an oral diet or after 4 weeks of treatment.All eases were evaluated videofluoroscopically with a drinking test pre-and pest-treatment.Results After 10 to 24 balloon dilata-tions,10 of the 15 patients in the treatment group regained the ability to take solid food and water orally,thoush 2 of them could take pasty food only.Only 2 of the 15 patients in the control group regained the ability to take common food by mouth,though 5 of them could take pasty food.The other patients had no improvement.There wag a signifi-cant difference between the two groups.The cricopharyngeal aehalasia of 12 patients in the ffeatment group improved from incomplete relaxation/opening to complete relaxation/opening.Pooling and residue in the pyriform sinus or val-leculae was reduced and no misaspiration was observed.In the control group only 7 patients had some improvement.The mesn time for the bolus passing the pharynx after treatment was significantly shortened from 0.23 s to 0.15 s in the treatment group,but not significantly in the control group.Conclusions Catheter balloon dilatation is effective for cricopharyngeal achalasia caused by brainstem injury and is helpful for relieving the symptoms in the pharynx phase and the esophagus phase of dysphagia.