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Optimization of purification technology for polyphenols from vitis amurensis seed extract with macroporous resins

Journal of Jilin University(Medicine Edition).2017;43(6):1272-1277. doi:10.13481/j.1671-587x.20170639

Objective:To explore the purification technology parameters for polyphenols from vitis amurensis seed extract with AB-8 macroporous resin and to establish the optimum purification process conditions,and to provide technical reference for further development and utilization of vitis amurensis seeds.Methods:The vitis amurensis seeds were regarded as raw material,the concentration of vitis amurensis seed polyphenols was detected, and the adsorption rate and desorption rate of polyphenols were calculated.The AB-8 macroporous resin was used to purify the polyphenols from vitis amurensis seed extract, and the purification technology parameters of vitis amurensis seed polyphenols were optimized by absorption-desorption experiment.The concentrations of polyphenol from vitis amurensis seed extract were compared between before and after purification, and the effect of the purification process was verified. Results: The AB-8 macroporous resin had good absorption and desorption properties to crude the extract of vitis amurensis seed polyphenols,and the adsorption rate and desorption rate were 90.48% and 71.42%.The optimum adsorption conditions and desorption conditions were as follows:the sample concentration was 10 g· L-1 ,the sample flow rate was 3 mL· min-1 ,the eluting concentration was 60%,the elution velocity was 2 mL·min-1 ,and the eluting volume was 2-fold column volume.Under this condition,the concentration of polyphenols from vitis amurensis seed extract after purification was increased from 35.02% to 88.8%.Conclusion:AB-8 macroporous resin has better purification effect,and it is worthy of popularization and application.

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Application of ultrasonography in diagnosis and differential diagnosis of Takayasu arteritis

Yu LIN ; Meihan LIU ; Weidong SHI ; Zhixia SUN ; Enqi CHEN ; Xin ZHANG ; Zhiyuan WANG

Journal of Jilin University(Medicine Edition).2017;43(6):1268-1271,后插3. doi:10.13481/j.1671-587x.20170638

Objective:To investigate the clinical application value of ultrasound in the diagnosis and differential diagnosis of Takayasu arteritis (TA ). Methods: A total of 39 suspected TA patients were examined by ultrasonography,erythrocyte sedimentation rate (ESR),C reactive protein (CRP)and CT angiography (CTA), and the ultrasound examination results were compared and analyzed with the final chinical diagnosis results. Results:Of the 39 cases of suspected TA patients which had been examined by ultrasound,33 cases were diagnosed as TA,2 cases were diagnosed as atherosclerosis (AS),2 cases were diagnosed as fibromuscular dysplasia (FMD) and 2 cases were diagnosed as thromboangiitis obliterans (TAO).Compared with final chinical diagnosis results, 1 FMD patient was misdiagnosed as TA,and the rest of the diagnostic results were consistent with the final chinical diagnosis results.Conclusion:Ultrasound examination can provide objective and accurate basis for the diagnosis and differential diagnosis of TA,and it is an effective method of imaging examination.

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Application of completely digital design technology in fabricating ceramic crowns to copy preoperative tooth:A case report and literature review

Qianyue SUN ; Qian LIANG ; Qianqian LI ; Shuang YANG ; Zhijian SHEN ; Zhe WU

Journal of Jilin University(Medicine Edition).2017;43(6):1253-1255,后插3. doi:10.13481/j.1671-587x.20170634

Objective:To fabricate an all-ceramic crown which copies the morphology of preoperative tooth in one patient with only small occlusal dentin defects with completely digital design (CDD)technology,and to find a more precise and feasible solution to fix the tooth detect.Methods:The tooth treated with root canal therapy had the complete axial surface and small occlusal dentin defects.In order to keep the morphology of preoperative tooth,the digitized crown was designed with the information of preoperative tooth by oral scanning with CS3500 and using the"Biogeneric Copy"function of exocad design software,then the digitized information of the teeth was collected and transformed and the simulate zirconia crown was mode by 3D colloidal deposition technique.The patient's comfort and satisfaction were investigated by observing the morphology,the interproximal contacts,and the occlusion of crown.Results:The morphology of definitive restoration was similar to the preoperative tooth,which presented good interproximal contact and occlusion,and the patient was greatly satisfied without discomfort and foreign body sensation.Conclusion: A more accurate and personalized restoration approach can be implemented with the application of CDD to fabricate new ceramic crowns which copy the preoperative tooth.

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Comparison of diagnosis value of MRI,CT and histopathological features on liposarcoma of extremities

Gang ZHU ; Haibin SUN ; Hu WANG ; Gang XU

Journal of Jilin University(Medicine Edition).2017;43(6):1215-1219,封2. doi:10.13481/j.1671-587x.20170627

Objective:To compare the detection results of histopathology and MRI, CT of liposarcoma of extremities,and to provide guide for increasing the accuracy rate of clinical diagnosis.Methods:The data of 43 cases of liposarcoma of the extremities were collected.MRI plain scan and enhanced CT were used in 18 patients before operation.Sixteen cases were diagnosed by CT plain scan and enhanced scan before operation.Nine cases were diagnosed by MRI and CT plain scan and enhanced scan before operation.All the patients underwent the operation. The intraoperative pathological examination was performed in the lesion tissue.The diagnosed results were assessed according to the classification (high differentiated liposarcoma, mucoid liposarcoma, toliposarcoma and mixed liposarcoma).Results:In all the 43 cases,the upper limb lesion was found in 13 patients and the lower limb lesion was found in 30 patients.The average maximal diameter of the lesions was (14.6±6.1)cm.The fat cells were found in all the patients,with the pathological characteristics of adipocytes and lipopolysaccharide mixture exists, and accompanied by deep-stained pleomorphic cells;the CT examination results showed that the low density shadow with clear boundary and linear separated shadow after CT enhanced;the MRI image showed short signal intensity on T1 weighted imaging and long signal intensity on T2 long signal and short time reversal recovery sequence imaging and other signals;the mass existed the cord-like,linear shadow;the lesion surrounding nerve and blood vessels were squeezed,but the bone structure was relatively complete.The CT and MRI results of well-differentiated lipsarcome showed the fat signal,and the enhanced results showed the mild strengthening.The CT and MRI results of mucinous liposarcoma showed the cystic density and signal, and the enhanced results showed the clound-like orlatticed mild and moderate strengthening;the CT and MRI results of mixed liposarcoma showed the mixed imaging findings of above two types.Conclusion:The pathological examination can provide the exact basis for the diagnosis of type of liposarcoma;CT and MRI imagings can provide basis for j udging the size of the tumor;MRI enhanced scan has an important role in judging tumor border.Combination of three kinds of detection methods can provide the basis for the diagnosis of the disease.

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Changes of serum levels of IGF-1,IGFBP-3 and plasma proteins of patients with diabetic foot before and after recombinant human growth hormone therapy and their significances

Tao SHANG ; Yanli REN ; Jing LI ; Ying ZHANG

Journal of Jilin University(Medicine Edition).2017;43(6):1209-1214. doi:10.13481/j.1671-587x.20170626

Objective:To detect the levels of serum insulin-like growth factor 1 (IGF-1),insulin-like growth factor binding protein 3 (IGFBP-3 )and the plasma proteins of the patients with diabetic foot (DF),and to investigate the therapeutic effect of recombinant human growth hormone (rhGH)in the DF patients.Methods:The DF patients (DF group,n=40),diabetes mellitus patients (DM group,n=40)and healthy adults (normal control group,n=40)were selected.The patients in DF group were divided into rhGH treatment group and conventional treatment group according to the treatment willingness (n= 20).The patients in rhGH treatment group recevied 0.4 IU·kg-1 ·d-1 rhGH injection subcutaneously for 15 d,the interval between two courses was 5 d,with a total of 3 courses of treatment.The patients in convention treatment group received subcutaneous injection of the same dose of saline.The general clinical data before and after treatment,the serum IGF-1 and IGFBP-3 levels, the constitution of disease grading of patients,the levels of plasma TP and TRF,FBG,and the efficiency rate and wound healing time of the subjects in DF group and normal control group were detected.Results:The levels of serum IGF-1 and IGFBP-3 of the patients in DF group were significantly lower than those in DM group and normal control group (P <0.01);the serum levels of IGF-1 and IGFBP-3 of the patients in DM group were significantly lower than those in normal control group (P <0.05).After treatment,the serum levels of IGF-1,IGFBP-3 and plasma TP,TRF of the patients in rhGH treatment group were significantly increased compared with before treatment (P <0.05),and they were higher than those in conventional treatment group after treatment (P <0.05). There was no significant difference in the FBG levels of the subjects between DM group and normal control group (P > 0.05 ).The effective rate of the patients in rhGH treatment group was higher than that in conventional treatment group (P <0.05).Conclusion:The efficiency of DF patients can be improved after treated with rhGH, and it is suitable for clinical promotion.

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Application of two dimension speckle tracking imaging in evaluating on cardiac dysfunction in patients with malignant lymphoma after anthracycline treatment

Fan ZHANG ; Yi ZHENG ; Xiaohong YANG ; Caifen YE ; Liangyu WANG

Journal of Jilin University(Medicine Edition).2017;43(6):1193-1198. doi:10.13481/j.1671-587x.20170623

Objective:To assess the changes of myocardial biomechanical parametres of left ventricle of the patients with non-Hodgkin lymphoma (NHL)after anthracycline (ANTH)treatment by using two dimension speckle tracking imaging (2D-STI),and to study the value of 2D-STI in evaluating and monitoring the early cardiac dysfunction of the NHL patients induced by ANTH treatment.Methods:A total of 37 hospital patients who were firstly diagnosed as NHL (ANTH chemotherapy group)and 20 healthy volunteers (normal control group)were selected.The global longitudinal strain (GLS),global radial strain (GRS),global circumferential strain (GCS) and left ventricle twist (LVtw)of the subjects in two groups were detected before chemotherapy and 1,2,3 weeks after chemotherapy with 2D-STI;the GLS×LVtw was calculated.Results:There were no significant differences in the general clinical parameters of the subjects between normal control group and ANTH chemotherapy group before chemotherapy (P >0.05);the values of GLS,GRS and GCS of the patients in ANTH chemotherapy group after ANTH chemotherapy were decreased than those in normal control group and before ANTH chemotherapy (P <0.05),and GLS had the most obvious change.The GLS, GRS, GCS and GLS × LVtw of the patients after 3 cycles of chemotherapy in ANTH chemotherapy group were decreased (P > 0.05 ). Four patients meet the diagnostic criteria of cardiac toxicity in the couse of chemotherapy.The sensitivity was 94%,the specificity was 66.7%,and the maximal Youden index was 0.667 when the △ GLS × LVtw =-64.53% ×°was used as the cut-off point in predicting the occurrence of myocardial toxicity.Conclusion:2D-STI can detect the early changes of biomechanical parameters of left ventricular myocardium with highly sensitive in predicting early myocardial toxicity and early cardiac dysfunction caused by ANTH chemotherapy.It may be an effective way to predict the early myocardial toxicity of ANTH chemotherapy in the future.

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Induction of oxaziridine on apoptosis of human cervical cancer HeLa cells and its mechanism

LI XU ; Wei ZHANG

Journal of Jilin University(Medicine Edition).2017;43(6):1161-1164. doi:10.13481/j.1671-587x.20170617

Objective:To explore the induction of oxaziridine on the apoptosis of human cervical cancer HeLa cells and its mechanism.Methods:The human cervical cancer HeLa cells were treated with 0,0.5, 1.0, 5.0 and 10.0 μmol·L-1 oxaziridine for 24 h and used as control group and different concentrations of experiment groups. The morphology of HeLa cells was observed under inverted microscope;the surival rates and inhibitroy rates of proliferation of the HeLa cells in various groups were measured by MTT assay.The HeLa cells were treated with 0, 0.5,1.0 and 5.0 μmol· L-1 oxaziridine for 12 h,and Hoechst 33258 staining was used to detect the apoptotic morphology of the cells;Western blotting method was used to detect the relative expression levels of Bcl-2 and Bax proteins.Results:The MTT assay results showed that the survival rates of the HeLa cells in 0.5,1.0,5.0 and 10.0 μmol·L-1 groups were decreased and the inhibitory rates of proliferation of the HeLa cells were increased (P <0.05 or P < 0.01 ) compared with control group, and the half inhibitory concentration (IC50 ) was 2.6 μmol·L-1. The cells in control group (0 μmol · L-1 oxaziridine group)were in good condition,and the volumes of the HeLa cells in 0.5, 1.0, 5.0 and 10 μmol · L-1 groups were significantly reduced and the intercellular junction also disappeared.The Hoechst 33258 staining results showed the enhanced staining in the cells treated with oxaziridine.The Western blotting results showed that compared with control group (0 μmol· L-1 oxaziridine group),the relative expression levels of Bcl-2 protein in the HeLa cells in 0.5,1.0 and 5.0 μmol·L-1 oxaziridine groups were significantly decreased (P <0.05 or P <0.01),and the relative expression levels of Bax protein were significantly increased (P < 0.05 or P < 0.01 ).Conclusion:Oxaziridine can effectively inhibit the proliferation of HeLa cells in vitro and induce the apoptosis of HeLa cells.

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Effects of cerebral ischemic postconditioning on expressions of Notch1,Cyclin D1 and CDK4 proteins in hippocampus of rats

Yao LIU ; Xiaoyun ZHAO ; Yaning ZHAO ; Jianmin LI ; Jianing LI

Journal of Jilin University(Medicine Edition).2017;43(6):1142-1147,后插1. doi:10.13481/j.1671-587x.20170614

Objective:To investigate the effects of cerebral ischemic postconditioning (CIP)on the expressions of Cyclin D1 and CDK4 proteins in hippocampal neurons of the rats with global cerebral ischemia and reperfusion (I/R)based on Notch1 signaling pathway,and to explore the mechanisms.Methods:A total of 128 healthy male SD rats were randomly divided into sham operation group,I/R group (modified Pulsinelli four vessel occlusion method), CIP group (repeated 3 times of reperfusion or blocking blood vessel before complete reperfusion)and DAPT group (intraperitoneal injection of 10 mg·kg-1 ·d-1 DAPT 3 h before CIP),and there were 32 rats in each group.HE staining was used to observe the morphology of the neurons at 6, 24, 48 and 72 h after ischemia;immunohistochemical staining was used to observe the expressions of Cyclin D1, CDK4 and Notch1 in the hippocampus of the rats;Western blotting method was used to observe the expression levels of Cyclin D1,CDK4 and Notch1 proteins in the hippocampus of the rats at different time points. Results:The HE staining results showed that compared with sham operation group,the structure of neurons in hippocampus area of the rats in I/R group was damaged and the survival rate of neurons was significantly decreased (P <0.05).Compared with I/R group,the survival rates of neurons in the hippocampus area of the rats in CIP group were significantly increased at the corresponding time (P <0.05).Compared with CIP group,the survival rates of neurons in hippocampus area of the rats in DAPT group at the corresponding time were significantly decreased (P < 0.05 ). The immunohistochemical staining results showed that compared with sham operation group,the number of cells with positive Notch1,Cyclin D1 and CDK4 in I/R group was increased (P < 0.05).Compared with I/R group,the number of Cyclin D1 and CDK4 positive cells in CIP group was decreased (P <0.05),and the number of Notch1 positive cells of was significantly increased (P <0.05).Compared with CIP group,the number of Cyclin D1 and CDK4 positive cells in DAPT group was increased (P < 0.05 ),and the number of Notch1 positive cells was significantly decreased (P < 0.05 ). The Western blotting results showed that compared with sham operation group,the expression levels of Notch1,Cyclin D1 and CDK4 proteins in the hippocampus of the rats in I/R group were increased (P <0.05);compared with I/R group,the expression levels of Cyclin D1 and CDK4 proteins in the hippocampus of the rats in CIP group were significantly decreased (P <0.05),and the expression level of Notch1 protein in the hippocanpus of the rats was significantly increased (P < 0.05 ); compared with CIP group,the expression levels of Cyclin D1 and CDK4 proteins in the hippocampus of the rats in DAPT group were increased (P <0.05),and the expression level of Notch1 protein was significantly decreased (P < 0.05).Conclusion:CIP may play an important role in protecting the neurons by increasing the activity of Notch1 and inhibiting the expressions of Cyclin D1 and CDK4.

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Effects of Ginkgo Bitoba extract on differentiation and bone resorption of osteoclasts and their mechanisms

Junmei WANG ; Qinyue SUN ; Zhe WU ; Tong LI ; Jing LI

Journal of Jilin University(Medicine Edition).2017;43(6):1130-1136,后插1. doi:10.13481/j.1671-587x.20170612

Objective:To study the effects of Ginkgo Biloba extract (GBE)on the differentiation and bone resorption of the osteoclasts,and to clarify their mechanisms.Methods:The RAW264.7 cells were cultured in vitro,then were treated with receptor activator for nuclear factor-κB ligand (RANKL)and different concentrations of GBE. The cells were divided into blank control group (0 μg · L -1 RANKL ), RANKL group (100 μg·L -1 RANKL),RANKL + 75 mg · L -1 GBE group, and RANKL + 150 mg · L -1 GBE group.The morphology and number of osteoclasts were assessed with TRAP staining assay,and bone resorption of GBE was examined with bone resorption pits assay. Flow cytometry was applied to analyze the the apoptotic rate of RAW264.7 cells and the cell cycle; the expression levels of nuclear factor of activated T cells 1 (NFATc1 ), DC-STAMP,Casthepin K,matrix metalloprotein 9 (MMP-9),Bcl-2,Bax,P27 and Cyclin-D1 in the RAW264.7 cells were analyzed by RT-PCR method.Results:Compared with blank control group,the number of osteoclasts in RANKL group were significantly increased (P <0.01);compared with RANKL group,the number of osteoclasts in RANKL+75 mg·L -1 GBE and RANKL+150 mg·L -1 GBE groups were significantly decreased (P <0.05). Compared with blank control group,the area of bone resorption pit of bone slice in RANKL group was significantly increased (P <0.05);compared with RANKL group,the areas of bone resorption pit of bone slice in RANKL+75 mg·L -1 GBE and RANK+150 mg·L -1 GBE groups were significantly decreased (P <0.05).Compared with blank control group,the apoptotic rates of the RAW264.7 cells in 75 and 150 mg · L -1 GBE groups were increased,and the expression levels of Bcl-2 in the RAW264.7 cells were significantly decreased and the expression levels of Bax were significantly increased (P <0.05).Compared with RANKL group,the G0-G1 phase arrest of the RAW264.7 cells in RANKL+ 75 mg· L -1 GBE and RANKL 150 mg· L -1 GBE groups were shortened;the expression levels of P27 in the RAW264.7 cells were significantly decreased and the expression levels of Cyclin-D1 were significantly increased (P < 0.05).Compared with blank control group,the expression levels of NFATc1, DC-STAMP,Casthepin K and MMP-9 in the RAW264.7 cells in RANKL group were significantly increased (P <0.05);compared with RANKL group,the expression levels of NFATc1,DC-STAMP,Casthepin K and MMP-9 in the RAW264.7 cells in RANKL+75 mg·L -1 GBE and RANKL+ 150 mg·L -1 GBE groups were significantly decreased (P <0.05).Conclusion:GBE could inhibit the differentiation and bone resorption of the osteoclasts,and their mechanisms may be related to promoting the apoptosis of RAW264.7 and shortening the G0-G1 phase of the RAW264.7 cells.

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Effect of velvet antler polypeptides combined with Schwann cells modified with GDNF gene on proliferation of human bone marrow mesenchymal stem cells in vitro

Bocheng LU ; Xiangyang LENG ; Pengcheng ZHANG ; Ying WANG ; Xukai WANG

Journal of Jilin University(Medicine Edition).2017;43(6):1125-1129,后插1. doi:10.13481/j.1671-587x.20170611

Objective:To investigate the effect of pilose antler polypeptide combined with Schwann cells modified by glial cell line-derived neurotrophic factor (GDNF)gene on the proliferation of human bone marrow mesenchymal stem cells (BMSCs)in vitro .Methods:According to the conventional method,the bone marrow (10 mL)was extracted from the healthy volunteers and was inoculated into the culture flask.The primary cultured cells were completely fused.The BMSCs were harvested at the 3rd generation and the cells were adjusted to 5 × 106 mL-1 . 4 μL GDNF gene modified Schwann cells was added into GDNF group,4 μL (10 mg· L-1 )PAP combined with GDNF gene modified Schwann cells was added into combination group,and only same amount of medium was added into control group.The proliferative activities,cell nuclear antigen (PCNA)levels and apoptotic rates of BMSCs in various groups were detected by enzyme-linked immunosorbent assay,ELISA method and Annexin Ⅴ-FIFC/PI cell apoptosis detection kit,respectively.Results:After primary culture for 48 h,most of the cells adhered to the wall,and the morphology of the cells changed into polygonal shape and few of them showed spindle.The passaged cells showed spindle spindle,the cells were confirmed as BMSCs,and all of them were non-hematopoietic stem cells.Compared with control group,the proliferative activities and the PCNA level of the BMSCs in GDNF group and combination group were increased (P <0.05)and the apoptotic rates were decreased (P <0.05);compared with GDNF group,the proliferative activity and the PCNA level of the BMSCs in combination group were increased (P < 0.05 )and the apoptotic rate was decreased (P < 0.05 ).Conclusion:PAP combined with Schwann cells modified by GDNF gene can promote the proliferation of human BMSCs in vitro .

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