[Objective]To study the relationship between trauma and the lumbar disc degeneration and to detect the expression of the fibronectin splicing variants in the degenerated lumbar intervertebral disc,and study the expression of EDA.[Method]The human lumbar intervertebral disc tissues were collected to extract total RNA which was then amplified by RT-PCR technique.[Result]EDA~+ Fn of normal lumbar intervertebral disc were not expressed,whereas EDA~+ Fn of degenerated lumbar intervertebral disc were expressed abundantly.The defference between different groups has statistical significance.[Conclusion]EDA~+ Fn has very strongly expression in the lumbar degenerated intervertebral disc,and the trauma is one of prerequisite factors in the disc degenerated process probablely.

To summarize and introduce the available methods of establishing rabbit models of VX2 nasopharyngeal carcinoma ( NPC) , and to explore the improvements at each stage in the preparation of the rabbit models, in order to provide a favorable animal model for future experimental research.

Objective To investigate the expression of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in hippocampus after status convulsion (SC) and explore the influence of age and duration of SC on the expression of BDNF and NGF. Methods Animal model with the different durations of SC (30 min,3h) were established by intraperitoneal injection of 3 mEq/kg lithium chloride,18-20 h later,followed by 5 mg/kg pilocarpine in 40 adult Wistar rats aged 2-3 months and 40 20-day-old Wistar rats. The normal control and experimental control were made in ten adult rats and ten 20-day-old rats (n=5 for each control). The rats were sacrificed at 3 h,6 h,12 h,1 d,3 d,7 d after 30-minute SC,or 3 h,1 d after 3-hour SC respectively. The location and cell type expression of BDNF and NGF in the hippocampus were observed by immunohistochemistry. The levels of BDNF and NGF at different time points were quantitatively analyzed by enzyme-linked immunosorbent assay (ELISA). Results The cells expressing BDNF and NGF located mainly in the dentate granule cells and CA1-CA4 pyramidal cells of hippocampus. The most positive immunoreactivity was in cytoplasm,partly in axons. The expression of BDNF had the tendency of increase in all rats after 3-hour SC,peaked at 1 d and 3 d respectively. These increases lasted for at least 7 d. Moreover,a several-fold increase was observed at the peak levels. The patterns of NGF expression were similar to that of BDNF after SC. The elevated degree and duration of NGF expression were remarkably lower than that of BDNF,declining abruptly to below control levels by 12 h with the characteristic of biphasic increase. The expression of hippocampal BDNF had the positive correlation with the duration of SC,but that of NGF was not. No matter the duration of SC,there was more rapid and evident induction of BDNF in 20-day-old rats and adult rats. Moreover,the age-related difference was more obvious for the longer duration of SC. Conclusion SC induced the expression of BDNF chiefly in hippocampus,and had minor influence on that of NGF. The expression of BDNF was related to age and duration of SC. There was a more rapid and marked induction of BDNF in 20-day-old rats and adult rats. The age-related difference had the positive correlation with the duration of SC.

Objective To explore the impacts of 75％ low-protein diet intake during gestation on fetal growth restriction (FGR) rat model establishment.Methods Thirty-eight pregnant Sprague-Dawley rats were included into the study.At first,five pregnant rats were fed with sufficient normal diet with protein content of 22％.Their daily food consumption was recorded and taken as the basis to determine daily feed consumption of 75％ low-protein group (protein content 9.2％).In order to ensure that each group finally had at least ten pregnant rats to deliver,there were 11 rats assigned to the control group (pregnant rats fed with sufficient normal diet,protein content was 22％),13 to the low-protein group (pregnant rats fed with low protein diet,protein content was 9.2％,but the food consumption was the same as control group) and 14 to the 75％ lowprotein group (pregnant rats fed with low-protein diet,protein content was 9.2％,the food consumption was 75％ of the control group).All female rats were fed with sufficient normal diet after delivery.The body weight,overall weight gain during gestation,the mortality rate and the non-delivery rate of pregnant rats were compared.The third day's newborn weight after birth,FGR incidence and the mortality rate within three days after birth of newborns were also compared.One way analysis of variance,LSD-t test,independent sample t-test and Chisquare test were used as statistical methods.Results (1) The body weight of pregnant rats:There was no significant difference in body weight among the three groups at gestational day 0,3 and 6.On day 9,body weight of 75％ low-protein group [(271.9±8.4) g] and low-protein group [(274.1 ±7.8) g] were significantly lower than that of the control group [(287.2± 18.7) g] (t=2.514 and 2.170,both P＜0.05),but there was no significant difference between the former two groups.On Day 12,body weight of 75％ low-protein group [(275.7 ± 10.7) g] and low protein group [(285.1 ± 12.5) g] were significantly lower than that of the control group [(306.4±29.7) g] (t=3.262 and 2.218,both P＜0.05),and the difference between the former two groups was also statistically significant (t=2.098,P＜0.05).Before delivery,body weight of 75％ low-protein group,low protein group and control group were (300.4±14.1) g,(317.0±16.3) g and (372.9±19.1) g,respectively with statisticall significance (F=64.219,P＜0.05).The overall weight gain during pregnancy for 75％low-protein group,low-protein group and control group was (61.6± 19.8) g,(81.8±21.6) g and (139.3± 12.0) g,respectively.The difference among the three groups was statistically significant (F=55.863,P＜0.05).(2) The mortality rates of pregnant rats for 75％ low-protein group,low-protein group and control group were 3/14,2/13 and 1/11 respectively without significant difference (P＞0.05).Neither was the non-delivery rate within 30 days (embryonic resorption) for the three groups (1/14,1/13,0/11,P＞0.05).(3) The numbers of pups were 101 in 75％ low-protein group,104 in low-protein group and 107 in control group.The newborn mortality rate within three days after birth was 28.7％ (29/101) in 75％ tow-protein group and 23.0％ (24/104)in low-protein group,with were significantly higher than that of the control group (7.5％,8/107) (x2=16.022and 9.976,both P＜0.05),but there was no significant difference between groups.The third day's newborn weight after birth for 75％ low-protein group,low-protein group and control group were (6.3 ±0.8) g,(6.9±0.9) g and (8.1 ±0.9) g,the difference was statistically significant (F=90.602,P＜0.05).FGR incidence for 75％ low-protein group was 55.6％ (40/72),which was significantly higher than that of the low-protein group (28.8％,23/80) and the control group (5.0％,5/99) (x2=11.220,54.834 and 18.833 all P＜0.05).Conclusion 75％ low-protein diet feeding during pregnancy is an ideal method to induce FGR rat model with high FGR incidence,whereas and low mortality rates of pregnant rats,the fetuses and newborns.

To discuss the availability of evaluation on the dissolution studies of the multicomponents in traditional Chinese medicine, the in vitro dissolution of total composition of the tablet of rhizomes of Ligusticum chuanxiong components and its correlation with the in vivo were studied by the method of area under the absorbance-wavelength curve (AUAWC). Taken the tablet of rhizomes of Ligusticum chuanxiong components which is composed of sodium ferulate and ligustrazine hydrochloride as subject model, the dissolution tests were carried out with basket method. The plasma concentrations of tablets in different rats were determined by AUAWC at different interval times. The in vivo absorption percentage was calculated by Wagner-Nelson equation to evaluate the in vitro and in vivo correlation. According to the results, the cumulative dissolution in vitro of total composition of tablets of rhizomes of Ligusticum chuanxiong components at 60 min was 90.65% in water by AUAWC. The in vivo pharmacokinetics is fitted with an one-compartment model. The linear equation based on the cumulative dissolution rate (fr) and absorption percentage (fa) at 5, 10, 20, 30 and 60 min was fa = 0.819 7 fr+0.183 and the correlation coefficient was 0.959 5, which showed a good correlation between the in vitro dissolution and the in vivo absorption percentage. The method of AUAWC can be used accurately, feasibly and conveniently to evaluate the in vitro and in vivo correlation of total composition of tablets of rhizomes of Ligusticum chuanxiong components, which will provide better guidance to study the in vitro and in vivo correlation of sustained release preparation etc under complex system of traditional Chinese medicine in the future.

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 mid-term outcomes for the application of homograft valve conduits in right ventricular outlfow reconstruction in patients with congenital heart disease. Methods: We retrospectively studied 122 patients who received right ventricular outlfow reconstruction by homograft valve conduits application in our hospital from 2007-10 to 2014-07. The patients were divided into different sets of groups, by surgical procedure: Ross group,n=38 and Non-Ross group,n=84; by median age: ≤6 years group,n=61 and >6 years group, n=61; by the type of valve conduits: Aortic homograft group,n=21 and Pulmonary homograft group,n=101; by the diameter of conduits: ≤19 mm group,n=31 and >19 mm group,n=91. The relationships between pre-operative conditions, different types of conduits and diameters to the prognosis were analyzed; the post-operative death, re-operation, free homograft valve conduits failure rates were followed-up in all patients. Results: The average follow-up time was (35.4 ± 22.2) months and 2/122 (1.6%) patients died during that period, the overall free conduits failure rates at 1, 5 and 7 years post-operation were 94.2%, 81.2% and 75.4% respectively. The free conduits failure rates in Pulmonary homograft group at 1, 5, 7 years post-operation were 96.2%, 86.1%,79.9% and in Aortic homograft group were 80.0%, 59.7%, 59.7% respectively,P=0.011; in Ross group were 96.4%, 89.0%, 89.0% and in Non-Ross group were 91.3%, 78.3%, 67.1% respectively,P=0.045. While the age, conduits diameter, cyanosis and re-operation had no statistical meaning to free conduits failure rates, allP>0.05. Conclusion: Application of homograft valve conduits had good mid-term outcomes in right ventricular outflow reconstruction in patients with congenital heart disease, while the long-term effects should be further emphasized in clinical practice.

Objective To establish an UPLC method for the determination of sinomenine in Sinomenine External Applied Powder. Methods The UPLC method was carried out on a C18 column by using acetonitrile-water-ethylene diamine (50:50:0.25) as mobile phase. The flow rate was 0.2 mL/min; the sample quantity was 2 μL; the detection wavelength was 283 nm. Results The peak time was within 1 min or so. The calibration curve of sinomenine was in the linear range of 34.2–2188.0 ng. Conclusion The method is simple, rapid, stable and reliable, which can be used for the determination of sinomenine in Sinomenine External Applied Powder.

Objective To evaluate the efficacy of splenectomy plus selective pericardial devascularization under endoscope in the treatment of advanced schistosomiasis patients with portal hypertension and hypersplenism so as to explore the minimally in-vasive and safer surgical treatment. Methods A secure splenectomy was performed with laparoscope and its supporting devices, and at the same time,the ligation of the left gastric vein and the ligation of esophageal vein perforating vertically into the esopha-gus were also performed in 14 advanced schistosomiasis patients with portal hypertension and hypersplenism. Results Among the 14 patients,the splenic artery was separated and clipped before the treatment of splenic pedicle. One patient was of conversion to open laparotomy for the splenic vein rupture bleeding in the separation. There was no death. Conclusion The operation of sple-nectomy plus selective pericardial devascularization under endoscope is effective,truly minimally invasive,and safe in the treat-ment of advanced schistosomiasis patients with portal hypertension and hypersplenism.

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.