OBJECTIVE: To investigate the relationship between syndrome differentiation of traditional Chinese medicine (TCM) and characteristic changes of vascular endothelial function in patients with diabetic arterial occlusion (DAO) of lower extremities. METHODS: Forty patients with DAO were selected as trial group. Twenty patients among them were attributed to blood stasis syndrome (group A1), and the others were attributed to syndrome of pathogenic dampness-heat attacking the lower limb (group A2) according to syndrome differentiation type of TCM. Patients with diabetes (group B), arteriosclerosis obliterans (group C) and healthy people (group D) were observed as the control groups, respectively. There were 20 cases in each group. Endothelium-dependent dilation (EDD) and endothelium-independent dilation (EID) were measured by high resolution ultrasound in the 100 subjects and the changes of vascular tension factors were also studied. RESULTS: The results showed that EDD in group A was reduced significantly as compared with that in the groups B, C and D. The levels of vascular contractile factors such as endothelin-1 (ET-1) and thromboxane B2 (TXB2) in group A were higher than those in the groups B, C and D, while the levels of vascular dilatory factors such as nitric oxide (NO) and 6-keto-prostaglandin F1alpha(6-Keto-PGF1alpha) were declined significantly as compared with those in the groups B and D. Linear correlation analysis showed that EDD was correlated positively with the levels of NO and 6-Keto-PGF1alpha, while the levels of ET-1 and TXB2 had negative correlation with EDD. EDD and EID in group A2 were declined significantly as compared with those in group A1. CONCLUSION: Our findings indicate that endothelial dysfunction may play an important role in the pathogenesis of DAO and may be associated with syndrome differentiation of TCM.

BACKGROUND: Cassia seed acts on decreasing blood pressure and blood lipid, protecting liver and inhibiting bacteria. It is worth to carry on a further discussion on its effect of weight loss.OBJECTIVE: To observe the influence of cassia seed decoction drunk naturally on body mass of nutritional obese rats in physiological state.DESIGN: Completely randomized grouping was designed, in which, control experiment, analysis of variance and q test were applied in comparison among groups.SETTING: Cardiovascular Institute, Second Affiliated Hospital, Henan University of Science and Technology.MATERIALS: The experiment was performed in Cardiovascular Instutute,Second Affiliated Hospital, Henan University of Science and Technology from March 2004 to September 2004, in which, 27 male SD rats were employed and randomized into 3 groups, named normal control group, model group and cassia seed group, 9 rats in each one.METHODS: [1] In normal control, the rats were bred with basic forage(the contents of protein, fat, carbohydrate were 18.2%, 4.5% and 55.2%successively, with 14.54 kJ caloric each gram) and drank water naturally.In model group, the rats were bred with high nutritive forage (the contents of protein, fat, carbohydrate were 23.7%, 21.6% and 39.0% successively,with 19.56 kJ caloric each gram) and drank water naturally. In cassia seed group, the rats were bred with high nutritive forage and drank cassia seed decoction of various concentration naturally. The concentration of cassia seed decoction started at 10 g/L (equally contained 10 mg raw cassia seed each milliliter) and was increased by 100% concentration each day (10 g/L)till to 60 g/L on the 6th day. Since the 7th day, the concentration of 60 g/L was maintained till to the 7th weekend. [2] It was to record appetite and drinking quantity at definite time every day and calculate absorbed caloric(intake mass × caloric contained each gram). It was to measure body mass at definite time each week. On the 7th weekend, the body length of rat was measured and Lee's index was calculated [ 3√body mass (g)×103/body length (cm)]MAIN OUTCOME MEASURES: Influences of cassia seed on body mass, Lee's index, appetite, caloric and drinking quantity in nutritional obese rats.RESULTS: Twenty-seven rats all entered result analysis. [1] Body mass:that in model group from the 3rd to 7th week in experiment group was higher remarkably than normal control group (P ＜ 0.05-0.01). That in cassia seed group from the 2nd to 7th week was lower remarkably than that in the model group (P ＜ 0.05-0.01). [2] Lee's index: that in model group and cassia seed group on the 7th week of experiment was higher remarkably than that in the normal control group [(358.60±8.55), (341.84±7.29), (322.00±6.89) g/cm, P ＜ 0.05-0.01] and that in cassia seed group was lower remarkably than that in the model group (P ＜ 0.05). [3] Appetite: that in model group and cassia seed group was lower remarkably than that in the normal control group (P ＜ 0.05-0.01) and that in cassia seed group was near to the control group (P ＞ 0.05). [4] Absorbed caloric: that in model group and cassia seed group was higher remarkably than that in the normal control group (P ＜ 0.05-0.01) and that in cassia seed group was near to the control group (P ＞ 0.05). [5] Drinking quantity: that in cassia seed group was basically near to that in the model group and the control group (P ＞ 0.05) and that in model group was near to the control group. It was indicated that cassia seed decoction at mass concentration of 60 g/L did not affect appetite.CONCLUSION: Cassia seed decoction at mass concentration of 60 g/L inhibits remarkably the increased body mass of nutritional obese rats and is free from influence on appetite.

Objective To study the early changes of the coagulation system in type 2 diabetic nephropathy.Methods Sixty-two cases of patients with type 2 diabetic nephropathy were divided into two groups:normal albuminuria group ( N-UAlb group,UACR ＜ 30 mg/g,32 cases ),microalbuminuria group ( MUAlb group,UACR:30～300 mg/g,30 cases).Thirty healthy persons constituted a control group (NC group).Fibrinogen( FIB ),antithrombin Ⅲ ( AT-Ⅲ ),protein C ( PC ),protein S ( PS ) were measured by coagulation analyzer,while yon willebrand factor (vWF) and platelet granule membrane protein 140 (GMP-140) were detected by ELISA assay,platelet count (PLT),mean platelet volume(MPV),platelet hematocrit (PCT),platelet distribution width(PDW) by hematology analyzer.Results The level of fibrinogen,GMP-140 and vWF in the M-UAlb group were (4.20 ± 1.53 ) g/L,( 30.03 ± 7.77 ) μg/L,and ( 315.53 ± 47.24 ) ％ respectively,vwhich were significantly higher than those in the N-UAlb group [ ( 3.21 ± 0.89 ) g/L,( 18.22 ± 5.08 ) μg,/L and ( 191.88 ± 57.25 ) ％ respectively ] and the NC group [ ( 2.75 ± 0.53 ) g/L,( 14.26 ± 2.29 ) μg/L and ( 138.12 ± 61.27 ) ％ respectively ] ( F =5.42,10.42,30.44,P ＜ 0.05 or 0.01 ).The fibrinogen,vWF,GMP-140 were positively correlated with UACR ( r =0.313,P ＜ 0.05 ; r =0.620,P ＜ 0.01 ; r =0.680,P ＜ 0.01 ) and PC was negatively correlated with UACR ( r =-0.255,P ＜ 0.05 ).Conclusion Hypercoagulable state in diabetic nephropathy is associated with the high fibrinogen,endothelial dysfunction and platelet activation,and these changes have already emerged in patients without albuminuria.This might mind us that we should strengthen anticoagulant therapy on patients when they are not progressing to albuminuria.

Objective To study the effects ofShenqi YizhiGranules (SQYZ) on learning and memory and content of Aβ1-42 of cerebral tissue in 5XFAD mice with Alzheimer’s disease; To discuss its mechanism on improving learning and memory ability of 5XFAD mice.Methods Four-month-old C57BL?6 wild type mice were randomly divided into NS control group and SQYZ control group, and the 5XFAD mice were randomly divided into model group, SQYZ group and huperzine-A (HupA) group, 15 mice in each group. Each group were given same volume for gavage for 60 d. After treatment, the learning and memory ability were evaluated by nesting test, passive avoidance and Morris water maze test. The senile plaques and content of Aβ1-42, ionized calcium binding adapter molecule 1 and glial fibrillary acidic protein in cerebral cortex and hippocampus were detected by immunohistochemical staining and immunofluorescence, respectively.Results Compared with NS control group, the score of nesting test in model group significantly decreased; the step-through latency in passive avoidance was shortened and the escape latentcy in Morris water maze test was prolonged; the quantity of senile plaques and content of Aβ1-42 increased in cerebral cortex and hippocampus; the activation of glial cells significantly increased. In the SQYZ group, the above-mentioned indexes reached or approached the level of wild type control mice. The difference between SQYZ group and model group was statistically significant (P<0.05,P<0.01).Conclusion SQYZ improved learning and memory ability in 5XFAD mice, which may be related to reduction of senile plaques, inhibition of over activation in glial cells and reduction of content of Aβ1-42 in cerebral cortex and hippocampus.

Objective: To study whether the bond strength between enamel and composite resin could be enhanced by intraoral sand abrasive. Methods: Ten human maxillary first incisior teeth were divided into 2 groups The experimental group was sandblasted with 30 ?m Al 2O 3 (CoJet Sand, pressure 300 kPa) from a distance of 5 mm for 5 seconds, and the control group were not sandblasted. The Herculite composite resin composite cylinders were bonded with Coltene system. Bonded specimens were stored in 37 ℃ distilled water for 24 h, then were subjected to shear force in a testing machine Stress at failure was calculated in Mpa, and mode of failure was recorded. The Student t test was applied to the data. Results: The shear bond strength of experimental groups was (33.0?1.8) MPa , and that of control groups was (26.7?5.2) MPa ,there was significant difference between these two groups. All the adhesive failures happened at the enamel composite resin interface , except that cohesive failure happened in one sandblasted specimen. Conclusion: Intraoral sandblasting could significantly enhance the shear bond strength between enamel and composite resin.

Objective Since the discovery of multiple bioactivities of agarobiose oligomers, the establishment of quantitative analysis of agarobiose oligomers obtained under different hydrolysis modes would be of much importance. Methods Agarobiose oligomers ranged from biose, tetraose, hexaose, octaose, to decaose were isolated and purified by gel chromatography on Sephadex column. The agarobiose oligomers were separated in RP-HPLC after introducing ?-naphthylamine into compounds. Results Each oligomer could be quantified with good linearity and the lowest detection limit reached 0.1～2 ?g?mL -1. The chromatograms profiles under different hydrolysis modes(HCl, citric acid, solid acid, and hydroxyl radical degradation)showed that the content of agarobiose could reach more than 57.8 % using solid acid hydrolysis with best yield of 33.2 % . While HCl could degrade agar into a series of agaro-oligosaccharides from biose to decaose. The yield of oligosaccharides was low if hydrated by citric acid. The method of Fenton degradation could increase the speed of hydrolysis, but the products were more complex. Conclusion The method established here could be useful for analysis of hydrolysates of oligomers under different hydrolysis modes.

Objective To construct the RNA interference(RNAi)recombinant adenovirus vector targeting at human hypoxia-inducible transcription factor 1?(HIF-1?)and to evaluate its effect on human lung adenocarcinoma cell line SPCA-1.Methods The recombinant adenovirus Ad was constructed.HIF-1? inserted with HIF-1? RNAi fragment via AdEasy system.The virus was purifed by CsCl gradient centrifuge.The functional titer of recombinant adenovirus was measured by transfection test in HEK 293 cells.SPCA-1 cells were transducted with 2 multiplicity of infection(MOI)Ad.HIF1? in vitro,the expression rate of green fluorescence protein(GFP)was recorded by flow cytometry,HIF-1? mRNA and protein level was measured by Real-Time RT-PCR and flow cytometry.ResultsThe recombinant shuttle plasmid PAdTrack.HIF-1? and adenovirus plasmid Ad.HIF-1? were all correct shown by enzyme digestion confirmation.The plasmid pAd.HIF-1? was transducted into HEK293 cells,15%GFP expressionwere seen after 3 days.The final titers of recombinant adenovirus were 5.0?1010 TU/mL.SPCA-1 cells was transducted by Ad.HIF-1? in vitro for 48 h,GFP expression rate was 92%,HIF-1? mRNA and protein level decreased 89% and 87%,respectively.Conclusion RNAi adenovirus vector of human HIF-1? gene has been successfully constructed,which could facilitate the research onHIF-1? gene related gene therapy for lung cancer.

Objective To evaluated the prevalence of EGFR mutations in Hubei region,to provide reliable experimental basis for reasonable screening TKI beneficiaries in clinic.Methods EGFR mutation of 253 patients diagnosed with NSCLC in Hu-bei region during 2010 to 2015 was detected by PCR-sanger sequencing and ARMS assay,to investigate the sensitivity of dif-ferent methods,to explore the frequency and clinical significance of EGFR mutation in different gender,in different histologi-cal type and different exons.Results 93 of 253 NSCLC patients harbored EGFR mutation,mainly occurred in exon 19 and 21,accounting for 53.76% and 35.38% of the total EGFR mutation rate,respectively.ADx-ARMS method showed higher sensitivity (P=0.001).The mutation detection rate of female NSCLC was significantly higher than that of male NSCLC (P=0.001).The observed incidence of EGFR mutations in patients with adenocarcinomas (38.01%)was the highest in differ-ent histological type,followed by glandular squamous cell carcinoma (30.77%),large cell carcinoma (20%)and squamous cell carcinoma (4.55%).Nonsmoking patients had a higher EGFR mutation frequency (51.6%,81/157)than those with a history of smoking (24%,12/50).Conclusion ARMS assay was more sensitive and more convenient detection method for clinical screening for EGFR-TKI treatment subpopulation.The mutation rate of female NSCLC was significantly higher than that of male NSCLC in Hubei Province.Speculated that the sexual differences in NSCLC with EGFR mutation frequencies were related to hormone levels and smoking status.

Objective To explore the effect of predepositautotransfusion in hematopoietic stem cell donors.Methods 127donors of hematopoietic stem cell wereretrospectively analyzed from 2013 to 2016,whenhematopoietic stem cells collection and autologous blood transfusion.Results Collection autologous blood 70 550 mL by 127 donors,and 2 cases had mild blood donation reaction,at a rate of 1.6％ (2/127).Collectionbone marrow stem hematopoietic stem cell 100 159 mL.The detection results were WBC (27.02±8.21) × 1010/L、Plt(213.19±62.42) × 109/L、RBC(4.31±0.44) × 1012/L、Hb(131.53 t14.53)g/L,from 117 donors beforespinal cord extraction.The detection results were WBC (35.8519.08)× 1010/L、Plt (188.34±50.24) × 109/L、RBC (4.02± 0.41) × 1012/L、Hb (122.60 ± 13.48) g/L,from 117 donorsafter autologous blood transfusion in one day.Compared with donors after autologous blood transfusion in one day,WBC was higher before spinal cord extraction(P＜0.05),Plt RBC and Hb was significantly decree (P＜0.05).Conclusion Preoperative autologous blood donation can save blood and prevent blood-borne infectious disease,which can used in Hematopoietic stem cell donor.