Objective To investigate the effect of different transdermal enhancers on percutaneous absorption of ephedrine in Compound Herba Ephedrae Adhesive Plaster.Methods Using modified Franz diffusion cell,the ephedrine content was determined to investigate the effect of different transdermal enhancers in promoting percutaneous absorption.Results Azone or azone used together with other transderml enhancers had a better effect in promoting percutaneous absorption,indicating that azone was the leading transderml enhancers for ephedrine.The mixed transdermal enhancer of azone and propylene glycol was the most effective combination and the rate constant of percutaneous absorption is 94.280 ?g?cm-2?h-1.Conclusion The mixture of azone and propylene glycol can enthance the percutaneous absorption of ephedrine and can be used as the transdermal enhancers for this compound plaster.

Objective To study on purication of stilbene glycoside and icarrin with macroporous resin of radix polygoni multiflori praeparata cum succo glycines sotae and herba epimedii of Fufang Bushenshengjing Capsule. Methods To select the best type of resin from NKA-9, AB-8, D101, X-5 and DM130, to observe the factor of the concentration of water-extraction, the volume of resin, pH of water-extraction, the concentration and volume of ethanol. Results Using AB-8 resin, the concentration of water-extraction is 0.3 g/mL, the volume of resin (mL) to the mass of medicinal materials is 2, pH of water-extraction need't change, eluant is 70% ethanol and its volume (mL) is 5 times of the mass of medicinal materials (g). Conclusion This research can provide reference for the extraction of active component from Fufang Bushenshengjing capsule in industrial production.

Objective To explore the effect of nasal continuous positive airway pressure ventilation on severe respiratory disease and hypoxaemia. Methods In 30 neonates with severe respiratory disease-induced hypoxemia were managed with nasal continuous positive airway pressure, and corresponding nursing. Nasal continuous positive airway pressure before and after treatment, 24h arterial partial pressure of oxygen (PaO2), partial pressure of carbon dioxide (PCO2), PaO2/FiO2 and pH value in children were observed. Result After nasal continuous positive airway pressure ventilation, the levels of PaO2, post-PCO2, PaO2/FiO2 and pH value of children were better than the positive pressure ventilation before treatment and positive pressure ventilation (P < 0.05). Conclusions Nasal continuous positive airway pressure ventilation has good therapeutic effect on severe respiratory disease in children with neonatal hypoxemia. Good ventilation pressure regulation, skin and mucosa protection, prevention of vomiting in children with nursing to ensure the positive significance of continuous positive airway pressure therapy.

objective To determine the existence of virulence-associated invA gene in different genospeeies of Leptospira interrogans reference strains in China.and to understand the alterations of invA gene transcription and expression of L.interrogans strain Lai before or after infecting cells.Methods PCR was applied to detect the invA gene of four L.interrogans strains belonging to four different genospecies and L.biflexa strain Patoc Ⅰ.The entire invA genes from the L.interrogans strains were cloned and then sequenced.The prokaryotic expression system of invA gene of L.interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai was constructed.Using Ni-NTA affinity chromatography,the target recombinant protein rInvA was extracted and purified.Rabbits were immunized with rInvA to obtain antiserum and the titer of antiserum was determined by immunodiffusion test.A model of L interrogans strain Lai infecting human embryo kidney cell line HEK293 was established to detect the alterations of invA gene transcription and expression of the leptospiral strain before or after infecting the host cells by real-time fluorescent quantitative RTPCR and western blot assay.Results All the four tested L.interrogans strains had invA gene whereas L.biflexa strain Patoc Ⅰ not.The similarity of nucleotide and putative amino acid sequences of invA genes from the four L.interrogans strains belonging four different genospecies were 99.33%-100%and 98.66%-100%,respectively.The constructed prokaryotic expression system could efficiently express rInvA and the immunodiffusion titer of rabbit anti-rInvA serum was 1:16.After L.interrogans strain Lai infecting HEK293 cells for 30 min or above,the microbe could adhere the surface of the cells.On the 30 min after the infection,the mRNA level of invA gene of L.interrogans strain Lai was remarkably upregulated,and on the 45 min after infection the mRNA level presented a peak value and then graduated decreased.On the 45 min or 60 min after L.interrogans strain Lai infecting HEK293 cells,InvA protein could be detectable but before infection or after infection for over 90 min the InvA protein expression was negative.Conclusion The invA gene is a unique gene for pathogenic L.interrogans.The invA gene of L interrogans has characteristics of cell-touched expression and transient expression,which may have a close correlation with adhering and invading host cells.

Objective To find out the optimal supercritical CO2 fluid extr ac tion prosess for Tetrahydropalmatine. Methods The extraction process was optimiz ed by orthogonal design with the yield of Tetrahydropalmatine as markers. Three factors including extraction pressure, temperature and the volume of entrainer were observed. Results The optimal condition of supercritical CO2 fluid extrac tion process was: under the pressure of 15Mpa at 40 ℃ , 1.5 fold of 95 % EtO H with Corydalis yanhusuo W.T.Wang as the entrainer. Conclusion The optimal supe rcritical process is feasible, with the advantage of low temperature and energy consumption, short time, high production and no organic residua.

Objective To investigate the expressions and relationship between lysine-specific demethylase 1 (LSD1) and E-cadherin protein in gastric cancer tissues and adjacent normal tissues,and the correlation with the clinicopathological features and prognosis of patients with gastric cancer.Methods The case-control study was adopted.The gastric cancer tissues and adjacent normal tissues were collected by surgical resection from 80 patients with gastric cancer who were admitted to the Xiangya Hospital of Central South University from June 2008 to June 2009.Expressions of LSD1 and E-cadherin protein were detected by immunohistochemistry (IHC).The follow-up of telephone interview was performed to detect survival of patients till June 2015.Relationships between the expressions of LSD1 and E-cadherin protein and clinicopathological features or prognosis of patients were analyzed.Comparison of count data and correlation were analyzed by the chi-square test and Spearman rank correlation analysis.Survival curve was drawn using the Kaplan-Meier method,and survival analysis was done using the Log-rank test.Results Expressions of LSD1 in cancer tissues and adjacent normal tissues were located at the cell nucleus.The positive expression rate of LSD1 was 67.5％ (54/80) and 43.8％ (35/80) in cancer tissues and adjacent normal tissues,respectively,with a significant difference (x2=9.141,P ＜ 0.05).Expressions of E-cadherin protein in cancer tissues and adjacent normal tissues were located at the cell membrane and cytoplasm.The positive expression rate of E-cadherin was 63.8％ (51/80) and 81.3％ (65/80) in cancer tissues and adjacent normal tissues,respectively,with a significant difference (x2 =6.140,P ＜ 0.05).The positive expression rate of LSD1 was 83.3％ (25/30),76.0％ (19/25) and 40.0％ (10/25) in the low-,moderate-and high-differentiated tumors,37.5％ (6/16),72.7％ (16/22),71.9％ (23/32) and 90.0％ (9/10) in the Ⅰ,Ⅱ,Ⅲ and Ⅳ stages of TNM stage,36.4％ (4/11) and 72.5 ％ (50/69) in patients without and with lymph node metastasis,respectively,showing significant differences (x2=12.870,9.425,4.111,P ＜ 0.05).The positive expression rate of E-cadherin protein was 53.3％ (16/30),56.0％ (14/25) and 84.0％ (21/25) in the low-,moderate-and high-differentiated tumors,75.0％ (12/16),63.6％ (14/22),71.9％ (23/32) and 20.0％ (2/10) in the Ⅰ,Ⅱ,Ⅲ and Ⅳ stages of TNM stage,100.0％ (11/11) and 58.0％ (40/69) in patients without and with lymph node metastasis,70.0％ (49/70) and 20.0％ (2/10) in patients without and with distant metastasis,respectively,showing significant differences (/x2 =6.494,10.073,5.547,7.426,P ＜ 0.05).There was a negative correlation in expressions between LSD1 and E-cadherin protein (r =-0.355,P ＜ 0.05).The survival time and 5-year overall survival rate of patients with positive and negative expressions of LSD1 were (36.9 ± 2.5) months and 31.1％,(47.4 ± 3.4) months and 56.0％,respectively,showing a significant difference (x2=4.550,P ＜0.05).The survival time and 5-year overall survival rate of the patients with positive and negative expressions of E-cadherin protein were (44.0 ± 2.5) months and 46.7％,(32.6 ± 3.5) months and 24.9％,respectively,showing a significant difference (x2 =7.306,P ＜ 0.05).Conclusions Positive expression of LSD1 in gastric cancer tissues is higher than that in adjacent normal tissues.There are increased expression of LSD1 and reduced expression of E-cadherin protein in low-differentiated gastric cancer tissues and high TNM stage,showing a negative correlation between them.Positive expression of LSD1 and negative expression of E-cadherin protein may indicate a poor prognosis.

Objective:To investigate the expressions of Smad2 and Smad4 in breast carcinoma tissue,and to analyze their relationships with oncogenesis and development of breast carcinoma and significances.Methods:Fifty-three samples of breast ductal carcinoma tissue and 50 samples of surrounding normal tissue were selected.The expression levels of Smad2 and Smad4 in cancer tissue and surrounding normal tissue were detected with immunohistochemical S-P method,and the relationships between the expression levels of Smad2 and Smad4 and the clinicopathologic parameters of breast carcinoma were evaluated.Results:The expression level of Smad2 protein in the breast carcinoma tissue was significantly higher than that in the surrounding normal tissue (z = - 2.08,P <0.05);the expression level of Smad4 protein in breast carcinoma tissue was lower than that in the surrounding normal tissue (z= - 5.01,P < 0.01).In breast carcinoma tissue,the Smad2 and Smad4 expressions were not significantly correlated with age (r=0.035,P >0.05;r=-0.077,P >0.05),tumor size (r= 0.128,P >0.05;r=0.133,P >0.05),lymph node invasion (r =0.163,P >0.05;r =0.006 P >0.05),distant metastasis (r =0.113,P >0.05;r = 0.126,P > 0.05),ER expression (r = 0.056,P > 0.05;r = 0.047,P > 0.05) and PR expression (r=0.129,P >0.05;r=0.107,P >0.05).However,the expression levels of Smad2 and Smad4 were negatively correlated with the expression of HER2 (r = - 0.388,P < 0.01;r = - 0.360,P < 0.01 ) and pathological grade (r = - 0.331,P < 0.05;r = - 0.388,P < 0.01 ).The expression of Smad2 was positively correlated to the expression of Smad4 in breast carcinoma (r=-0.83,P <0.01).Conclusion:The expressions of Smad2 and Smad4 may play an important role in the development of breast carcinoma,and they may be used as the potential biological markers for evaluating the degree of malignancy and prognosis of breast carcinoma.

Objective To investigate the effect of long-term proton pump inhibitor on osteoporosis in elderly patients.Methods A total of 150 patients with peptic ulcer treated in our hospital from January 2011 to January 2015 were selected as the observation group.150 healthy subjects were selected as the control group.The age,height,body weight and PPI time of the two groups were recorded.The changes of bone mineral density before and after treatment were measured by bone mineral density analyzer,ineluding lumbar L1-4,radial density and ulna density.The changes of bone mineral density were observed and recorded in the observation group before treatment,six months,1 year and 2 years after treatment.Results After treatment,the levels of gastrin were significantly increased in the observation group,and the serum calcium concentration and bone mineral density were significantly decreased (P＜0.05).The density of lumbar vertebrae,radius and ulna was significantly lower in observation group than those of control group (P＜0.05).With the prolongation of PPIs,lumbar vertebrae,radius and ulna density in observation group showed a decreasing trend.Conclusion Long-term application of proton pump inhibitors in elderly patients can cause bone loss.

Objective To investigate the clinical phenotype and genotype in three probands with antithmmbin(AT)deficiency and their families,and to identify the molecular mechanism of AT deficiency.Methods Chromogenic substrate method and immunoturbidimetry assay was used to detect the plasma levels of AT:A and AT:Ag,respectively.Genomic DNA was extracted from the peripheral blood.All 7 exons and the flanking sequences were amplified by PCR.and the abnormal mutant genes were analyzed by direct sequencing.Western blot was used to detect the AT levels and thrombin generation tests were used to detect coagulation status.Results The plasma levels of AT:A and AT:Ag of the three probands declined by 50%.G7386C(Trp225Cys)mutation in exon 4,C2591G(Ser36stop)in exon 2 and C9819T(Arg359stop)in exon 5 were characterized in the three prebands and they could result in W(Trp)225C(Cys)missense mutation,S(Set)36X(stop)nonsense mutation and R(Arg)359X(stop)nonsense mutation respectively,The testing results of phenotype and genotype from some of their family members showed consistent with results from the probands.Western blot results indicated that the Icyels of PC:Ag were lower compared with the normal pooled plasma.The hypercoagulative status was present in the probands using thrombin generation tests.Conclusions Type Ⅰ hereditary AT deficiency was found in these three families.The 3 heterozygous mutations.W225C,S36X and R359X are genetic defects of hereditary AT deficiency.W225C and S36X have not been described before.

Objective To explore the effects of SB203580(a MAPK inhibitor) on the expression of Homer1a in hippocampus and learning-memory after diffuse brain injury in rats.Method Male Sprague-Dawlley rats were divided randomly into three groups:control group,diffuse brain injury(DBI) group and DBI+ SB203580 group (peritoneal injection,0.01 μg/kg).Morphological changes of neuronal cells were observed by electron microscope and the expression of Homer1a and phosphorylated p38MAPK was detected by immunohistochemistry and learning and memory functions were performed with Morris water maze (MWM).Results Compared with control group,ultrastructure of neuronal cells and synapses were significantly.The levels of phosphorylated p38MAPK(76.98±16.64,2.28±0.40,P＜0.05) and Homer1a (62.96± 12.74,1.28±0.10,P＜0.05)respectively were increased after injury impaired.MWM test showed that the escaping latency was prolonged((74.64± 8.96)s,(24.96±4.98)s,P＜0.05) and the frequency of crossing the platform was decreased(4.48± 1.12,12.65±2.36,P＜0.05).Compared with the model group,SB203580 decreased ultrastructure impariment in neuronal cells and synapses and decreased phosphorylated p38MAPK expression(54.82± 12.48,76.98± 16.64,P＜0.05) and increased Homer1a expression(54.82 ±12.48,76.98± 16.64,P＜0.05).MWM test showed that the escaping latency was shorten ((46.72±6.58) s,(74.64± 8.96) s,P＜0.05),and the frequency of crossing the platform was increased (7.56± 1.20,4.48± 1.12,P＜0.05).Conclusion SB203580 improves the learning-memory recovery after DBI,which is related to inhibition of p38MAPK activation and increasing Homer1a expression.