1.Real-time Study of Interaction between Adenosine Triphosphateand Its Aptamer Using Dual Polarization Interferometry
Tao HU ; Fan YANG ; Xiurong YANG
Chinese Journal of Analytical Chemistry 2017;45(7):944-950
In this work, we used dual-polarization interferometry to explore the binding events between adenosine triphosphate (ATP) and ATP-binding aptamer (ABA) at solid-liquid interface.The single-stranded ABA was immobilized onto the chip surface.After the addition of ATP, the real-time and label-free technique for detailed investigation of their interactions were reflected on the changes of the mass, thickness, and density through DPI.By analysis of the binding curves from changes in mass, the association rate constant (ka, 4.66×10.3 L/(mol·s), dissociation rate constant (kd, 1.70 × 10.-2 s.-1), dissociation constant (KA, 2.7 × 10.5 L/mol), and association constant (KD, 3.7 × 10.-6 mol/L) were precisely determined.Moreover, good linear correlations between ATP concentrations and three parameters (mass, thickness, density) resolved by the response to ATP binding were obtained.The detection limits (LOD, 3σ) were 0.22 μmol/L for mass calibration, 0.14 μmol/L for thickness calibration, and 0.32 μmol/L for density calibration.We expect that this DPI-based aptasensor can be utilized to study the interactions of functional ABA with ATP, and also can be used for the detection of ATP with high sensitivity.
3.UTILIZATION OF PROTEIN IN VEGETARIAN AND MIXED DIET BY HUMAN ADULTS
Wenhsun FAN ; Wenchuan HU ; Enfu YANG
Acta Nutrimenta Sinica 1956;0(03):-
Vegetarian and mixed diets, supplying about 3000 Cal. and 8,5 gms. of protein daily, were given alternately to 6 adult men of 22-34 years old. In the vegetarian diet, composed of cereals, legumes and vegetables, about 20% of the total protein were supplied by the legumes. In the mixed diet, the legumes and part of the cereals of the above were replaced by meats, eggs and milk, furnishing about 50% of the total protein. The experiment consisted of 5 consecutive periods of 10 days each in the first three periods and 5 days each in the last two.Urinary nitrogen excretion of the experimental subjects during the period on vegetarian diet was lower than that on mixed diet. When the diet was changed from vegetarian to the mixed ration, urinary nitrogen remained low in the first two days and increased from the third day. However, when the change of diet was reversed, there was an immediat'e decrease of nitrogen excretion. The nitrogen balance was in favour of the vegetarian diet.The essential amino acid contents of the vegetarian diet were lower than that of the mixed diet. Therefore, better utilization of the protein in the vegetarian diet cannot be satisfactorily explained by essential amino acid index. With adequate caloric intake, 85 gms. of protein supplied daily either by vegetarian or by mixed diet may maintain nitrogen balance and protein storage in adult man.
4.Comparative study on intraoperative neuromonitoring of recurrent laryngeal nerve during thyroidectomy
Bo HU ; Hua YANG ; Fan YANG ; Xueqing JIANG
Journal of Endocrine Surgery 2013;7(1):11-13
Objective To investigate the value of intraoperative neuromonitoring(IONM) for preventing recurrent laryngeal nerve(RLN) injury in thyroid surgery.Methods 1087 consecutive patients undergoing thyroid surgery at Wuhan Central Hospital from Oct 2010 to Dec 2012 were evaluated.277 cases(the study group) underwent thyroidectomy with RLN identification by naked eye plus IONM.810 cases(the control group)underwent thyroidectomy with RLN identification by naked eye only.Results In the control group,733 RLNs (90.49%,733/810)were successfully identified and 77 RLNs were failed to be identified.In the study group,all the 277 RLNs(100%,277/277)were successfully identified.28 cases had postoperative temporal RLN injury,among whom 26 cases were in the control group and 2 cases were in the study group.18 cases in the control group and 2 cases in the study group recovered in 2 weeks after surgery.The rest recovered in 2 months after surgery.RLN injury rate was significantly lower in the study group than in the control group.There was no significant difference in RLN injury for low-risk surgical patients between the 2 groups; however,the study group had advantages in RLN injury than the control group for high-risk patients.Conclusions IONM in thyroid surgery can improve the recognition rate of RLN during thyroidectomy.IONM can significantly reduce the incidence of RLN injury,especially in high-risk surgery.
5.A prospective cohort study on the influence of high doses of herbs for clearing heat and resolving stasis on survival rates in patients with hepatitis B-related acute-on-chronic liver failure.
Xiaoyu HU ; Yang ZHANG ; Guo CHEN ; Sen ZHONG ; Xinjian FAN
Journal of Integrative Medicine 2012;10(2):176-85
Hepatitis B-related acute-on-chronic liver failure (ACLF) is a severe clinical syndrome characterized by jaundice, coagulopathy, ascites and hepatic encephalopathy and with a high mortality rate of 65% to 93%. It involves significant ethical issues when a randomized, double-blinded, placebo-controlled clinical study is conducted to such a serious disease. Therefore, a prospective cohort study design was utilized to explore a new treatment modality of applying integrated traditional Chinese and Western medicine.
6.8-Isoprostane in Exhaled Breath Condensate of Patients with Asthma
Ao HU ; Shaoying LI ; Weikang YANG ; Yongxia LI ; Manqi FAN
Chinese Journal of Respiratory and Critical Care Medicine 2009;8(4):337-340
Objective To invesitgate the relationship between 8-iseprostane (8-iso-PG) level in exhaled breath condensates (EBCs) and severity of asthma and explore the role of 8-iso-PG in asthma evaluation and monitoring.Methods Fifty-nine patients with asthma were enrolled.In which 15 eases were acute exacerbation, 13 eases were mild intermittent, 15 eases were mild persistent, and 16 eases were mederate-to-severe persistent.Thirteen healthy volunteers were recruited as control.EBCs were collected using EeoSereen system.The 8-iso-PG levels in EBCs were measured by a specific enzyme immunoassay.The patients with mild intermittent asthma were treated with inhaled corticosteroid (ICS) for one month and their EBCs were recollected for 8-iso-PG measurement.Results Exhaled 8-iso-PG levels were obviously increased in the patients with acute asthma compared with those chronic asthmatics [(47.2±6.8) pg/mL vs (24.5±12.0) pg/mL, P < 0.01].In the chronic persistent asthma, the levels were significantly higher in patients with mild persistent and moderate-to-severe asthma [(17.9±1.2) pg/mL and (39.7±4.0) pg/mL, P <0.01].While 8-iso-PG level did not differ significantly in intermittent asthma [(13.5±1.1) pg/mL] compared with the control subjects (P > 0.05).After one-month ICS treatment the 8-iso-PG level in the patients with mild intermittent asthma did not change significantly although the ACT score improved.Conclusions 8-iso-PG levels in EBC are associated with the severity of asthma,implicating 8-iso-PG may be useful in monitoring airway oxidative stress in asthma.ICS treatment is incapable of decreasing the 8-iso-PG,suggesting the ICS has minor impact on oxidative stress.
7.Experimental study on inhibitory effects of diallyl sulfide on growth and invasion of human osteosarcoma MG-63 cells.
Youzhen, HU ; Ling, CHEN ; Chengzhi, YI ; Fan, YANG ; Jige, CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(4):581-5
The inhibitory effects of diallyl sulfide (DAS) derived from allicin on in vitro and in vivo proliferation of human osteosarcoma MG-63 cells and the action mechanism, and the influence of DAS on invasive capability of MG-63 cells were investigated in order to search for the novel medicines for osteosarcoma. In the in vitro experiment, MG-63 cells were treated with different concentrations of DSA, and the morphological changes of MG-63 cells were observed under an inverted phase microscope. MTT method was used to assay the proliferation of MG-63 cells. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to detect the VEGF mRNA expression level in MG-63 cells. By using Transwell invasion assay, the influence of DAS on invasive ability of MG-63 cells was tested. In the in vivo experiment, the nude mice MG-63 cells tumor-bearing model was established, and different concentrations of DAS were injected beside the tumor. Twenty-one days after treatment, the mice were killed, the tumor size and tumor inhibition rate were calculated. The microvessel density (MVD) was determined by using immunohistochemistry. In the in vitro experiment, different concentrations of DAS could obviously inhibit proliferation of MG-63 cells in a time- and concentration-dependent manner. RT-PCR revealed that the expression levels of VEGF mRNA in DSA groups (different concentrations) were significant reduced as compared with those in control group (all P<0.05). Transwell invasion assay indicated that in 20 and 40 μg/mL DAS groups, the number of migratory cells was 91.4±8.3 and 81.8±7.4 respectively, which was significantly declined as compared with that in control group (150.4±14.7, both P<0.05). In the in vivo experiment, DAS could significantly suppress the growth of MG-63 tumor-bearing tissue. Immunohistochemistry demonstrated that different concentrations (20 and 40 μg/mL) of DAS could significantly decrease MVD of MG-63 tumor-bearing tissue (all P<0.05). It was suggested that DAS could inhibit the growth of MG-63 cells probably by suppressing the expression of VEGF mRNA.
8.Reactive hyperplasia of glial cells induced by spinal cord injury in a rat model
Xuhui FAN ; Bo YANG ; Xiang HU ; Fangxia GUAN
Chinese Journal of Tissue Engineering Research 2016;20(40):6001-6006
BACKGROUND:The nervous reconstruction and repair after spinal cord injury have become a research hotspot.
OBJECTIVE:To investigate the change rule of neurogliocyte reactive hyperplasia after spinal cord injury.
METHODS:Forty-two adult male Sprague-Dawley rats were selected and equivalently randomized into seven groups:normal control group (no intervention), sham operation group (lamina decompression) and operation groups (postoperative 1, 7, 14, 21 and 28 days). After the establishment of spinal cord injury models, the rats were sacrificed at each corresponding time point. The functional recovery of the rat hind limbs was evaluated by Basso, Beattie and Bresnahan scores, and complete spinal cord tissue was removed to undergo hematoxylin-eosin staining, immunohistochemistry staining and immunofluorescence staining.
RESULTS AND CONCLUSION:(1) Basso, Beattie and Bresnahan scores showed that rats in the normal control and sham operation groups had normal neurologic function. Rats at 1 day after spinal cord injury paralyzed completely, the neurologic function of hind limbs began to recover gradual y at the 7th day, and the recovery became most obvious at the 14th day, which had no significant differences compared with the 21st and 28th days. (2) Hematoxylin-eosin staining found that the diffuse hemorrhage and neuronal necrosis were observed in the injured area at 1 day after operation;inflammatory cel infiltration and some vacuoles appeared at the 7th day, and the hemorrhage was absorbed gradual y;the hemorrhage disappeared completely and capsule cavity formed at the 14th day;up to the 28th day, spinal cord structure was completely destroyed and that was replaced by cicatricial tissue accompanying with a large cavity. (3) Immunohistochemistry staining showed that the astrocyte in damaged area proliferated with the cel synapse increasing, which was most overt at the14th day;the axon clearance widened and the structure was in disorder at the 7th day, and the myelin sheath in the damaged area was destroyed at the 21st day. (4) Immunofluorescence staining showed that there were numerous visible glial fibril ary acidic protein+/nestin+cel s in the injured area at 14 days after operation. (5) These results suggest that glial cel hyperplasia and hypertrophy, the up-regulated expressions of glial fibril ary acidic protein and nest protein are advantageous to the early repair of spinal cord injury.
9.TiO2-coated Hollow Fiber Micro-Extraction Combined with Electrothermal Vaporization-Inductively Coupled Plasma Mass Spectrometry for Trace Elements Analysis in Environmental Water Samples
Fan WANG ; Man HE ; Beibei CHEN ; Yang WANG ; Bin HU
Chinese Journal of Analytical Chemistry 2015;(9):1313-1321
TiO2 coated hollow fiber ( HF) was prepared by sol-gel method and characterized by X-ray powder diffraction and scanning electron microscope. The adsorption performances of the self-prepared TiO2 coated HF for interest metal ions were explored. The sample solution was extracted for 30 min at pH 8. 0 under stirring at speed of 700 r/min, then desorpted with 100μL of 1 mol/L HNO3 . On the basis of this, a method combining TiO2 coated HF micro-solid phase extraction with electrothermal vaporization-inductively coupled plasma mass spectrometry ( ICP-MS ) was developed for the determination of trace metal ions in environmental water samples. Under the optimized conditions, the limits of detection obtained by the proposed method were 0. 039, 0 . 021 , 0 . 009 and 0 . 018 ng/mL for CrⅢ, CuⅡ, CdⅡ and PbⅡ with enrichment factors of 12 . 5 , 11 . 7 , 10. 3 and 18. 6, respectively. The preparation reproducibility of self-prepared TiO2 coated HF ranged from 4. 5% to 6. 8% (n=9) in one batch, and from 7. 7% to 9. 6% (n=7) in batch-to-batch. The developed method has been validated by analyzing a certified reference material ( GSBZ50009-88 200925 ) and also applied to the analysis of CrⅢ, CuⅡ, CdⅡ and PbⅡ in East Lake water and snow water successfully. Meanwhile, TiO2 coated HF stir bar was prepared for a comparison and it was demonstrated that TiO2 coated HF displayed higher extraction efficiency and adsorption capacity for target ions.
10.Efifcacy of the consolidation chemotherapy after autologous stem cell transplantation in refractory/relapse lymphoma
Yang YU ; Xing FAN ; Ling WANG ; Wei TANG ; Jiong HU
China Oncology 2014;(10):761-764
Background and purpose: High-dose chemotherapy followed by autologous stem cell transplantation (auto-HSCT) is considered as the ifrst line treatment for patients with relapse/refractory lymphoma after conventional chemotherapy. However, most of these patients still relapse the second time. The purpose of this study was to analyze the efifcacy of the consolidation chemotherapy after autologous stem cell transplantation (HSCT) refractory/relapse lymphoma in high risk. Methods:A total of 38 patients with relapsed/refractory lymphoma including Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL) were included, who were underwent auto-HSCT in our transplan-tation department from Jan. 2010 to Dec. 2013. In treatment group, 19 patients received 2 courses of consolidation che-motherapy after auto-HSCT every 2 to 3 months, with the regimen of mini-BEAM or modiifed mini-CBV. Another 19 patients had no chemotherapy after auto-HSCT as control group. Results:The median follow-up duration was 17.2 and 7.5 months in the treatment and control group respectively. The follow-up data demonstrated prolonged progression-free survival (PFS) in the treatment group than the control group [24.7 months vs 7.8 months, P=0.029 under intend-to-treat analysis ITT;24.7 months vs 5.2 months, P=0.01 under per protocol analysis(pp)]. There is also a trend of improved overall survival (OS) in the treatment group (P=0.055, ITT). Conclusion:Consolidation chemotherapy after auto-HSCT for refractory/relapsed lymphoma patients delay the relapse and tend to improve the overall relapse rate.