Objective To detect the proliferation and the expression levels of downstream target genes of Notch pathway of T-ALL CCRF-CEM(CEM)cell line treated with exogenous DLK1 protein,in order to investigatethe effects of DLK1 protein on the Notch pathway in CEM cells.Methods CCK-8 assay was performed to examine the proliferation of CCRF-CEM cells,which were treated with various concentration(0.5,1.0,1.5 μg/ml)DLK1 for various time(24,48,72 h).RFQ-PCR was applied to assess the mRNA expression level of Notch1 receptor and downstream target genes of Notch pathway in CEM cells,which were treated for various time(24,48,72 h).Results DLK1 protein stimulated the proliferation of CCRF-CEM cells,and the proliferation rates with different concentrations of DLK1 were 0.14±0.03,0.17±0.04,0.55±0.01 in 72 hours,respectively,there were statistic differences between that in the experimental group and that in the control group(P＜0.05).DLK1 protein up-regulated the Notch1 receptor and its downstream target genes HES1,c-myc and NF-κB.The relative transcript levels of target genes HES1 in 72 hours,c-myc in 48 hours and NF-κB in 72 hours were 3.2551±0.3100,1.6086±0.0941,2.0515±0.3453 respectively,and there were statistic differences between that in the experimental group and that in the control group(P＜0.05).Conclusion DLK1 protein stimulates the proliferation of T-ALL CCRF-CEM cells by up-regulating Notch1 receptor and downstream target genes HES1,c-myc and NF-κB of Notch pathway.

BACKGROUND:Epidural scar after laminectomy is one important reason for the secondary spinal stenosis, and local application ofSanqi sodium hyaluronate gel can prevent epidural scar adhesion after laminectomy. OBJECTIVE: To study the effects ofSanqisodium hyaluronate gel on α-smooth muscle actin expression in the process of rabbit's epidural scar formation. METHODS: In this study, there were ninety-six rabbits which were randomized into four groups and given 0.5 mL normal saline, 0.5 mLSanqi concentrated solution, 0.5 mL sodium hyaluronate and 0.5 mLSanqisodium hyaluronate gel around the dura. At 1, 2, 4, 8 weeks after treatment, immunohistochemistry staining was employed for analysis of α-smooth muscle actin expression. RESULTS AND CONCLUSION:At the end of weeks 1 and 2, the expression of α-smooth muscle actin antibody in the normal saline group was significantly higher than that in the other three groups (P < 0.01 orP < 0.05), but there were no significant differences among the Sanqi, sodium hyaluronate andSanqisodium hyaluronate gel groups (P> 0.05). At weeks 4 and 8, the expression of α-smooth muscle actin antibody in theSanqi sodium hyaluronate gel group was significantly lower than that in the other three groups (P < 0.01 orP < 0.05), and there was no significant difference among the latter three groups (P > 0.05). These findings suggest thatSanqi sodium hyaluronate gel can inhibit the expression of α-smooth muscle actin, and thus ease scar contracture.

Objective To explore the differences of curative effect and short-term prognosis to severe traumatic brain injury patients with three different early postoperative nutritional supports.Methods A retrospective study was performed on 60 severe traumatic brain injury patients received in Neurosurgical Intensive Care Unit of Northern Jiangsu People's Hospital from July 2014 to July 2016.A total of 60 cases were randomly divided into the early enteral nutrition support therapy group,the early parenteral nutrition group,and the early compound nutrition group.The clinical indicators such as basic clinical characteristics before treatment,the nutrition data in two weeks,the length of stay in the Neurosurgical Intensive Care Unit,complications and GCS scores between the three groups were observed and analyzed.Results The indicators of early compound nutrition group were fasting blood-glucose (5.74 ± 0.64) mmol/L,prealbumin(203.80 ± 10.45) mg/L,total serum protein(61.99 ± 1.34) g/L,blood hemoglobin (114.53 ± 2.69) g/L,C-reactive protein(0.37 ± 0.06) mg/dl.The length of stay in Neurosurgical Intensive Care Unit was (11.6 ± 0.42) days in the compound nutrition group while those in the early enteral nutrition group was (13.20 ±0.42) days and those in the early parenteral nutrition group was(14.65 ± 0.42) days.The postoperative complications of the compound nutrition group were significantly lower than other two groups.The GCS scores of early compound nutrition group was(11.40 ± 1.60),which was the best in three groups.The differences were statistically significant (P ＜ 0.05).Conclusions Early compound nutrition support has an exact curative effect on postoperative severe traumatic brain injury patients in Neurosurgical Intensive Care Unit.It can obviously improve the nutrition status of patients with less complications,shorter length of stay in Neurosurgical Intensive Care Unit,higher safety and lower degree of coma,worth clinical promotion.

Objective To optimize extraction technology of Fufang-Fuqing-Xiji.Methods Orthogonal test combined with Plackett-Burman experimental design was adopted to study the influence factors of water extraction in Fufang-Fuqing-Xiji. The contents of chlorogenic acid, paeoniflorin and caffeic acid in Fufang-Fuqing-Xiji were determined by HPLC gradient elution method at the same time. A multi-index comprehensive score method was applied to analyze the data and to screen the optimum extraction technology. Results The best extraction conditions of Fufang-Fuqing-Xiji were as following: threaded 80 mesh sieve, soaked 2 h, decocted 3 times with 12 times the amount of water for 1h each time.Conclusions This optimized extraction technology was rational and stable, which could be used for the extraction ofFufang-Fuqing-Xiji.

Objective To improve early diagnosis of testicular torsion,reduce misdiagnosis and reduce the rate of orchiectomy.Methods The diagnosis and treatment of testicular torsion in 31 cases were retrospectively ana-lyzed.Results 54.8% misdiagnosis rate was in this group,all the 31 cases were diagnosed by color Doppler ultra-sound,including 19 cases of retained testicle and orchiectomy in 12 cases (38.7%).In 19 cases of this group retained testis,testicular torsion time within 5 hours was 2 cases,and postoperative had testicles survival.In the 6 cases of testicular torsion time within 5 hours to 10 hours,5 cases had the testis survival.In the 5 cases of torsion of testis time was 10 hours to 24 hours,3 caseshad the testis survival.In the 6 cases of testicular torsion time more than 24 hours,2 cases of testis survival.After postoperative following-up,19 cases of retained testis had no recurrence,all the 31 cases were found no contralateral testicular torsion and all cases sex hormone levels were in the normal levels. Conclusion Testicular torsion is easily misdiagnosed,color doppler ultrasound should be preferred.Early diagnosis and aggressive surgical exploration,unity and strictly control the orchiectomy surgery indications,means a lot to reduce the rate of orchiectomy.

This paper discusses the mechanism of cellobiose in fungal cellulase induction a nd repression, and its inhibition of cellulases hydrolytic activity. Depending on the research result of cellulose binding domain, our hypothesis is that the main function of Exo-1,4-?-glucanase is to destroy th e crystal structure of cellulose to facilitaty hydrolyzing of ?-1,4 glucosidic bonds. A new strategy for the efficient transformation of cellulose material is advanced at t he end.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Hep G2 Cells ; Humans ; Interleukin-6 ; metabolism ; Liver Neoplasms ; metabolism ; pathology

AIM: To investigate the CPU86017 and its enantiomers inhibit abnormal gene expression of calcineurin and NFκB in rat cardiomyopathy induced by L-thyroxin and compare the effect of CPU86017 (racemate) with its 4 enantiomers: (7S, 13R), (7S, 13S), (7R,13S), and (7R,13R)-CPU86017 in this model. METHODS: The animals were randomly divided into 7 groups. The rat hypertrophied model was produced by treatment with L-thyroxin 0.2 mg·kg-1·d-1, sc for 10 d and treated with CPU86017 or its enantiomers 4 mg·kg-1·d-1, sc from d 6 to d 10. The changes in left ventricular (LV) weight index, redox system, and the NO and iNOS activity in the myocardium were investigated. The expression of mRNA of calcineurin、NF-κB in the left ventricle was measured. RESULTS: There were significant cardiac hypertrophy and oxidative stress in rats treated by L-thyroxin. The expression of calcineurin, NFκB mRNA were upregulated (P<0.05, compared with that of control). After treatment with CPU86017 (racemate and enantiomers), LV remodeling and the redox system were improved. CPU86017 and (7S,13R)-CPU86017 showed a better improvement on LV remodeling and the redox than the other isomers and restored the normal expression of calcineurin, NF-κB (P<0.05, P<0.01), respectively. CONCLUSION: It suggested that an up-regulation of calcineurin and NFκB possibly related to the altered intracellular calcium handling system plays a role in the progression of L-thyroxin induced cardiomyopathy and CPU-86017 and its 7S,13R-CPU86017 enantiomer effectively inhibit the abnormal expression of calcineurin and NFκB genes, the NOS enzyme and oxidant stress in the cardiomyopathy.

Objective To investigate the expression difference of vascular endothelial growth factor (VEGF)between lung cancer and lung benign disease,and its relation to clinical characteristics and prognosis of non-small cell lung cancer(NSCLC),and to research the possible role of VEGF in the growth of lung can- cer.Methods The expression of VEGF and microvascular density(MVD)were detected in 60 lung cancer tissues and 30 lung benign disease tissues after operation by immunohistochemical method and put up statisti- cal analysis.Results The positive rate of VEGF expression(63%)and MVD(45.13?10.27)in lung cancer tissues were both higher than those in lung benign disease tissues(27%,33?6.49)(P

Objective Insulin autoantibody (IAA) is known to exist in sera of type 1 diabetes mellitus (T1DM) patients and pre-T1DM individuals. The aim of this study was to establish a novel microtiter plate radioimmunoassay (RIA) for IAA and evaluate its clinical value. Methods Diluted 125Ⅰ-insulin was mixed with 5 ul serum samples in a 96-well microtiter plate and then incubated for 72 h on an orbital plate shaker (4℃). The immunocomplexes were transferred to another protein a coated Millipore plate, and then the plate was washed with Tri-Buffered Saline Tween-20 (TBT) buffer. Counts per minute (CPM) was measured with liquid scintillation and luminescence counter. The positive cut-off point of IAA index was defined as ≥0.06 based on the 99-percentile of the distribution in 317 healthy individuals. The specificity and sensitivity of the assay were calculated from the samples provided by the fourth Diabetes Autoantibodies Standardization Program (DASP 2005). The IAA levels were determined in 71 T1 DM and 551 newly diagnosed type 2 diabetes (T2DM) patients, and 317 healthy controls. The t test, non-parametric test, x2 test and linear correlation analysis were performed on the data using SPSS 11.5 software. The concordance rate was estimated with Kappa value. Results (1) The optimized testing condition was described as 2×104 CPM of 125Ⅰ-insulin, 5 ul serum sample and slowly horizontal shaking for 72 h. (2) The intra-assay CV was 4.8%-8.9% and inter-assay CV was 6.4%-10.5%. Based on DASP 2005 samples, the specificity and sensitivity of the assay were 97% (97/100) and 50% (25/50), respectively. Ninety-six serum samples with different IAA levels were selected and tested to compare between our new method and a domestic IAA RIA kit. The results showed that the IAA indices from the two methods were positively correlated (r= 0.678, P<0.001). The concordance rate was 72.9 %(Kappa value=0.402). There were 25 samples with discordant results, which were positive for IAA titer using the corresponding microtiter plate RIA but negative using the novel RIA kit. (3) In TIDM group the positive rate of IAA was 19.7% (16/71), higher than the healthy controls (0.9%, x2=54.36, P<0.001). The subgroup of T1DM children (with 0-9 years) showed the highest IAA positive rate (55.6% ,x2=4.85, P<0.05). In T2DM group the frequency of IAA was 1.5% (8/551), which had no significant difference comparing with that of healthy controls (x2= 0.95, P >0.05). Conclusions Our proposed microtiter plate RIA method for IAA is highly sensitive and specific, likely to be feasible for clinical application. The frequency of IAA is high in children with T1DM.