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2017 Vol. 47, No. 12 Published: 2017-12-25   859 Effect of puerarin on pancreatic β-cell injury in type 2 diabetic rats CHEN Xiufang, LEI Kangfu, DONG Min, ZHENG Qiaomin Objective: To study the effects of puerarin on diabetic pancreatic β-cell injury and to explore its possible mechanisms in type 2 diabetic rats. Methods: A type 2 diabetic rat model was established by the combination of a high-fat diet and a single low-dose streptozocin intraperitoneal injection. The animals were randomly divided into diabetic model group and puerarin (100 mg•kg-1, i.p.) treatment group, with other 8 normal rats as a control group. The rats were treated daily for 4 consecutive weeks, and the equal volume of vehicle (5% propanediol, i.p.) was given to the normal control and diabetic model group. The rats were sacrificed and fasting blood glucose, glycated hemoglobin (HbA1c) and serum insulin levels, as well as malondialdehyde (MDA) content, the activities of superoxide dismutase (SOD) and catalase (CAT) in pancreas were measured. The pancreatic telomere length was determined by real-time quantitative PCR. The protein levels of phosphorylated AMP-activated protein kinase (AMPK), sirtuin1 (SIRT1) and uncoupling protein 2 (UCP2) as well as apoptosis-related proteins, e.g. cleaved caspase-3/caspase-3, Bim, Bax and Bcl-2 in pancreas were assessed by Western blot. Results: The fasting blood glucose, HbA1c, and MDA contents as well as the protein expressions of UCP2, Bax, Bim and cleaved caspase-3/caspase-3 in pancreas of diabetic rats were significantly higher than those in normal controls (P<0.01, respectively). However, insulin level in serum, SOD and CAT activities, telomere length as well as the protein expressions of p-AMPK/AMPK, SIRT1 and Bcl-2 in pancreas of diabetic rats were significantly decreased as compared with those in normal animals (P<0.01, respectively). In contrast, in puerarin treatment group, the above changes were reversed, significant differences of those were found as compared with those in diabetic model group (P<0.01, respectively). Conclusion: Puerarin exerts preventive and remedial effects on the diabetic pancreatic β-cell, which is probably due to protecting telomere length and inhibiting β-cell apoptosis via alleviating oxidative damage, and decreasing UCP2 expression through enhancing SIRT1 activity as well. 2017 Vol. 47 (12): 859-863 [Abstract] ( 647 ) HTML (1 KB)  PDF (1678 KB)  ( 851 ) 864 the roles of N-acetyl galactosyltransferase-6 in proliferation and migration of breast cancer cell MCF-7 ZHOU Wenyuan, REN Jun, JI Yuan, CHEN Xiaoming. Objective: To explore the effects of N-acetyl galactosyltransferase-6 (GALNT6) on the abilities of proliferation and migration in MCF-7 cells. Methods: Generated sh-GALNT6 to knockdown the  GALNT6 expression in MCF-7 cells. The CCK8 assay, plate clone formation assay, cell cycle test and would healing assay were performed to detect the proliferation and migration activity of MCF-7 cells. The expression of PCNA, cyclinD1 and MUC1 as a downstream of GALNT6 were measured by Western blot. Results: The stable transfected sh-GALNT6 MCF-7 cell lines were constructed successfully. Down-regulation of GALNT6 could inhibit the proliferation and migration of MCF-7 cells. The expressions of PCNA, cyclinD1, and MUC1 were reduced in cells after they were transfected with sh-GALNT6. Conclusion: GALNT6 might play important roles in proliferation and migration of breast cancer, and it is also a potential molecular therapeutic target. 2017 Vol. 47 (12): 864-868,874 [Abstract] ( 730 ) HTML (1 KB)  PDF (2945 KB)  ( 907 ) 869 Preparation vitamin B2 modified multiwall carbon nanotubes electrode and determination of DNA by electrochemistry method ZHANG Tiantian, YANG Lizhu. Objective: The research on interaction between vitamin B2 which was on the vitamin B2/multiwall carbon nanotube modified electrode and DNA by electrochemistry and UV spectra methods was performed. Methods: There was an obvious pair of reduction and oxidation peaks on the prepared vitamin B2/multiwall carbon nanotube modified electrode. Both reduction and oxidation peak currents of vitamin B2 decreased with no change in the peak potentials after adding DNA. Through the test of the electrochemical parameters in the absence and presence of DNA, it was found that after combination of vitamin B2 and DNA under such condition, an electrochemically inactive supramolecular compound arised. Results: Under the experimental conditions selected, the decrease of oxidation peak currents had a linear relationship with DNA concentrations in the range of 0.01 to 0.1 mg/mL and 0.1 to 0.5 mg/mL, which could be used for measuring DNA concentration. Conclusion: The developed method is simple, rapid, economical and has good stability and reproducibility. 2017 Vol. 47 (12): 869-874 [Abstract] ( 608 ) HTML (1 KB)  PDF (1688 KB)  ( 1131 ) 875 Effect of curing time on cytotoxicity of Vitrebond DU Ruoxi, ZHOU Qiaozhen, YANG Zhengyu, MA Jianfeng. Objective: To investigate the influence of curing time of curing light on the cytotoxicity of Vitrebond, and compare the cytotoxic sensitivity of Agar diffusion test and MTT assay. Methods: Vitrebond of 2 mm thickness were light-cured for 15, 30, 45 seconds with 1 000 mw/cm2 curing light. The effects of different curing time of Vitrebond on L929 cell were evaluated by Agar diffusion test and MTT assay after 24 h and 48 h. Results: The evaluation of agar diffusion found the positive control and the specimen without any curing induced severe cytotoxity, there was no cytotoxicity in negative control, any of specimens and any of MEM or cottonseed oil extracts. In MTT test, Vitrebond showed a great cytotoxicity whatever the curing time was. There was no statistical difference of cytotoxicity cured for 15, 30, and 45 seconds (F=1.452, P>0.05). Conclusion: There are no difference of cytotoxicity of Vitrebond cured with 1 000 mw/cm2, with different curing time. Agar diffusion test is lower sensitive than MTT test to detect the cytotoxicity. 2017 Vol. 47 (12): 875-879 [Abstract] ( 616 ) HTML (1 KB)  PDF (1607 KB)  ( 903 ) 880 Inhibition effect of oridonin on human umbilical vein endothelial cells TIAN Lili, ZHU Guofu, SHENG Donglai. Objective: To study the inhibition effect of oridonin on human umbilical vein endothelial cells. Methods: The research chose human umbilical vein endothelial cells (HUVEC): cell viability was assessed by MTT assay, cell apoptosis was assessed by FACS analysis, cell migration and invasion was assessed by trans well chambers and image analysis, and cell angiogenesis was also investigated using tube formation assay. The expression of VEGFA and VEGFR2 were measured by real-time fluorescent quantitative PCR. Results: Oridonin induced HUVEC apoptosis and inhibited the proliferation, migration, invasion, and tube formation (P<0.05). The expression of VEGFA and VEGFR2 were down-regulated (P<0.05). Conclusion: Oridonin can inhibit HUVEC, which maybe connected with VEGFA and VEGFR2 down-regulation expression. 2017 Vol. 47 (12): 880-883,887 [Abstract] ( 652 ) HTML (1 KB)  PDF (2901 KB)  ( 856 ) 884 Combination of core neuromuscular facilitation technique and core stability training for rehabilitation of motor impairment after brain injury LI Haiyan, XU Leyi, HU Jinrong, LIN Xiaoke, WU Chao, ZHOU Chengye, ZHONG Yunjian. Guide::  Objective: To study the rehabilitation effect of the core neuromuscular facilitation (PNF) combined with core stability training on motor impairment in patients with brain injury. Methods: Fifty cases of brain injury patients in the First Affiliated Hospital of Wenzhou Medical University from June 2014 to October 2016 were randomly divided into the controlled group and the treated group with 25 cases in each, the controlled was given with PNF, and the treated was given with PNF combined with core stability training, but the same method was taken which was one treatment for 30 min, 1/d, 6 d/week and 8 weeks total. The efficacy of motor function was evaluated by Berg balance scale (BBS), trunk control ability test (TCT), maximum walking speed (MWS) and modified Barther index (MBI) before and after treatment respectively. Results: After treatment BBS, TCT, MWS of the two groups significantly improved, the difference was significant (P<0.05), no significant improvement of MBI (P>0.05); the two groups were compared with each other after treatment, BBS, TCT and MWS of the treated group were significantly higher than the other (P<0.05 MBI), no significant difference was found between two groups (P>0.05). Conclusion: The core stability training and PNF technology can improve the motor function of patients with brain injury, and the combination of the two techniques is better. 2017 Vol. 47 (12): 884-887 [Abstract] ( 663 ) HTML (1 KB)  PDF (1457 KB)  ( 1086 ) 888 The protective effect of curcumin derivative L6H4 on the hepatic tissue from type 2 diabetic rats DONG Xidan, CHEN Xiaohu, MIAO Chengfeng, XU Feifei, CHEN Sanmei, CAI Guoping. Guide::  Objective: To investigate the protective effect and its mechanism of curcumin derivative L6H4 on hepatic tissue from type 2 diabetic and hyperlipidemic rats. Methods: Forty male Sprague-Dawley (SD) rats were randomly divided into five groups evenly, normal control group, high fat group, high fat+L6H4 treated group, diabetes mellitus group and diabetes mellitus+L6H4 treated group. The rats were fed with high fat and high sugar diet to induce hyperlipidemic model, and then injected with streptozotocin intraperitoneally to induce type 2 diabetic models. Rats in high fat+L6H4 treated  group and diabetes mellitus+L6H4 treated group were given a gavage of curcumin derivative L6H4 at a dosage of 0.2 mg•kg-1•d-1 for 8 weeks. After the treatment, blood glucose and lipid levels as well as serum liver function indexes: alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB) were measured by automatic biochemical analyzer, free blood insulin (FIN) was tested by radioimmunoassay and homeostasis model assessment of insulin resistance (HOMA-IR) were calculated. The morphology of liver was observed under light microscope and transmission electron microscope. The expression of transforming growth factor β1 (TGF-β1), tissue inhibitor metalloprotemase 2 (TIMP-2) and matrix metalloproteinase 2 (MMP-2) were measured by immunohistochemical method. Results: Compared with normal control group, blood glucose and lipid levels, ALT as well as AST were significantly increased while ALB was significantly decreased in high fat and diabetes mellitus groups. After the treatment of L6H4, blood glucose and lipid levels as well as serum liver function indexes (ALT, AST) were markedly decreased while ALB was increased in high fat+L6H4 treated group and diabetes mellitus+L6H4 treated groups. Fatty degeneration and mild proliferation of fibrous tissue could be observed in high fat group, and the diabetes mellitus group showed further damage with focal necrosis, more pronounced fatty degeneration, and destruction of the hepatic cord, proliferation of fibrous tissue was remarkable, resulting in the thicker septa and the well observed deposition of collagen fiber in portal area, and were relived by the treatment of L6H4. The expression of TGF-β1, TIMP-2 and MMP-2 were markedly increased in high fat group and diabetes mellitus group, after the appliance of L6H4, TGF-β1 and TIMP-2 were decreased while MMP-2 was increased. Conclusion: Curcumin derivative L6H4 shows an effect in attenuating insulin resistance and regulating glucolipid metabolism, and exerts protective effect on the hepatic tissue from type 2 diabetic rats, down-regulation of the expression of TGF-β1, TIMP-2 and up-regulation of MMP-2 maybe involved in it. 2017 Vol. 47 (12): 888-894 [Abstract] ( 716 ) HTML (1 KB)  PDF (1966 KB)  ( 1008 ) 895 Effect of the chronic hypoxic-hypercapnia in apoptosis and activity of caspase-3 and p38 MAPK of hippocampus in mouse REN Huiming, WANG Xiaotong, YUAN Hai, KANG Hanlin. Objective: To investigate the effect of the chronic hypoxic-hypercapnia in apoptosis and activity of p38 MAPK and caspase-3 of hippocampus in mouse. Methods: Twenty-four male mice were randomly divided into two groups: the hypoxic-hypercapnia 4 weeks (4HH) group and normal control (NC) group. The 4HH groups in the special cantainer were exposed to an atmosphere containing 9%～11% O2 and 5%～6% CO2 8 hours a day, 6 days a week for 4 weeks. The two groups lived in the same room in the rest time. Apoptosis index (AI) in hippocampus was counted by TUNEL staining. Caspase-3 activity was measured by a caspase-3 activity assay kit. p-p38 MAPK protein in hippocampus was measured by Western blot. Results: Compared with NC group the AI, caspase-3 activity, and the p-p38 MAPK protein expression in hippocampus were significantly increased in 4HH group (P<0.01). Conclusion: The chronic hypoxic-hypercapnia promotes apoptosis and the activation of caspase-3 and p38 MAPK of hippocampus in mouse. 2017 Vol. 47 (12): 895-897 [Abstract] ( 646 ) HTML (1 KB)  PDF (1506 KB)  ( 784 ) 898 Antibacterial activity of polymyxin B in combination with several antimicrobial agents against carbapenem-resistant Klebsiella pneumoniae in vitro WU Zhenghai, LIU Haiyang, CHEN Huale, DUAN Darong, ZHOU Tieli. Objective: To evaluate the combined inhibitory effect of polymyxin B with meropenem and rifampicin respectively against carbapenem-resistant Klebsiella pneumoniae (CRKP) strains in vitro. Methods: Combined inhibitory effect of polymyxin B and meropenem, polymyxin B and rifampicin in vitro was assayed by detecting the minimum inhibitory concentration and calculating the fractional inhibitory concentration index for 20 CRKP strains with the checkerboard dilution method. Results: Combined  polymyxin B and meropenem showed 55% synergic effect, 45% additive effect, no indifferent and antagonistic effect on CRKP strains. Combined polymyxin B and rifampicin displayed 65% synergic effect, 35% additive effect, no indifferent and antagonistic effect on CRKP strains. Conclusion: Combined polymyxin B and meropenem, combined polymyxin B and  rifampicin show a combined synergic and additive inhibitory on CRKP strain in vitro. 2017 Vol. 47 (12): 898-901 [Abstract] ( 1158 ) HTML (1 KB)  PDF (1538 KB)  ( 4712 ) 902 Effect of anti-inflammatory dredging oviduct decoction on EGFR and ICAM-1 levels in rats with chronic salpingitis DAI Chunxiu, ZHUANG Xiaoping, YE Wanchun, YE Ren. Objective: To discuss the effect of anti-inflammatory dredging oviduct decoction on EGFR and ICAM-1 levels in rats with chronic salpingitis. Methods: Using mixed strains inoculation method, rat model with chronic salpingitis was manufactured. After making those models, those rats were divided into normal group, model group, rhizoma smilax china group and anti-inflammatory dredging oviduct decoction groups with different doses. The immunohistochemical and real-time pcr methods was used to observe effect of anti-inflammatory dredging oviduct decoction on EGFR and ICAM-1 levels in rats with chronic salpingitis. Results: Compared with normal group, the model group and the others all increasingly expressed EGFR and EGFR mRNA, and there were significant differences between results of two groups (P<0.05). Compared with model group, anti-inflammatory dredging oviduct decoction with the medium and high dose lowerd levels of EGFR and EGFR mRNA, both significantly decreased (P<0.05). Compared with normal group, the model group and the others all increasingly expressed ICAM-1 and ICAM-1 mRNA, and there was significant difference between results of two groups (P<0.05). Compared with the model group, anti-inflammatory dredging oviduct decoction with the medium and high dose lowerd levels of ICAM-1 and ICAM-1 mRNA, both significantly decreased (P<0.05). Conclusion: Anti-inflammatory dredging oviduct decoction can exert  therapeutic effects via inhibiting excessive expression of EGFR and ICAM-1 protein in oviduct epithelial cell. 2017 Vol. 47 (12): 902-905 [Abstract] ( 622 ) HTML (1 KB)  PDF (1565 KB)  ( 849 ) 906 The relationship between different surgical regimens and the prognosis of patients with organized chronic subdural hematoma TU Ming, ZHANG Zibin, WU Jinsen, SU Zhipeng, ZHU Danhua, ZHENG Weiming. Objective: To explore the relationship between different surgical regimens and the prognosis of patients with organized chronic subdural hematoma (OCSDH), and to evaluate contributing factors influencing surgical outcome. Methods: Clinical data of patients with OCSDH in the First Affiliated Hospital of Wenzhou Medical University from April 2011 to November 2013 were collected retrospectively. A total of 4 patients with OCSDH were identified and divided into 2 groups according to the two different operation methods, burr-hole drainage followed by craniotomy for hematoma removal, or direct-craniotomy for hematoma removal. Comparison between the two groups in terms of their preoperative and postoperative consciousness, GCS, physical examination, imaging, and the different surgical methods were compared. The relevance of membranectomy of OCSDH envelope firmly attached to the cerebral cortex, to prognosis of disease was also analyzed. Results: Patients who underwent membranectomy of OCSDH envelope had poor prognosis after surgery, one death and the other showed upward gaze and motor aphasia; the group with no resection of the envelope during surgery had better prognosis and neurological function. Conclusion: For OCSDH with envelope attached to cerebral cortex, we recommend the use of craniotomy for hematoma removal, without membranectomy of OCSDH envelope firmly attached to the cerebral cortex, since that results in to the better prognosis. 2017 Vol. 47 (12): 906-909,913 [Abstract] ( 675 ) HTML (1 KB)  PDF (2872 KB)  ( 813 ) 910 Analysis of factors affecting the uterine atony which isn’t sensitive to uterotonic agents in cesarean section SUN Rongrong, ZHANG Wenmiao, XIAO Biru, CHEN Qiuyue Objective: The purpose of this article was to identify risk factors and etiologiesleading to the uterine atony which wasn’t sensitive to uterotonic agents in cesarean section. Methods: The information of 705 patients who underwent caesarean delivery were collected between January 2014 and July 2016, who were divided into two groups based on the sensitivity of the uterotonic agents. Patients who were insensitive to uterotonic agents named group A (n=84, and the others named group B, n=621). Risk factors and etiologies for the uterine atony which wasn’t sensitive to uterotonic agents were explored using univariate and multivariate logistic regression analyses. Results: Multiple pregnancy (OR=6.495, 95%CI=3.801-11.098), adherent placenta (OR=2.865, 95%CI=1.595-5.144), placenta praevia (OR=4.096, 95%CI=1.912-8.660), general anesthesia (OR=1.470, 95%CI=1.198-1.803) were independently associated with the uterine atony which wasn’t sensitive to uterotonic agents. Conclusion: Multiple pregnancy, adherent placenta, placenta praevia and general anesthesia are the related factors of the uterine atony which isn’t sensitive to uterotonic agents should be pay attention to, we should take positive measures for the prevention and treatment. 2017 Vol. 47 (12): 910-913 [Abstract] ( 700 ) HTML (1 KB)  PDF (1451 KB)  ( 762 ) 914 Clinical effect of in situ bone chip replantation in the prevention of intracranial infection after external ventricular drainage XU Longbiao, ZHAO Ming, FANG Naicheng, PAN Bailin, DU Guosen. Objective: To investigate the clinical effect of in situ bone chip replantation in the prevention of intracranial infection after external ventricular drainage. Methods: A total of 156 cases in our department for external ventricular drainage in patients was randomly divided into 2 groups, replantation group (study group, 81 cases) and conventional replantation group (control group, 75 cases). In the study group of lateral ventricle puncture after the success of collected bone chips evenly in the drainage tube and the bone hole clearance in control group during the operation after successful puncture with gelatin sponge placed in the drainage tube and the bone hole clearance, the incidence of 2 groups of patients with cerebrospinal fluid leakage, intracranial infection and other complications were compared. Results: The incidence ratio of the research group of intracranial infection (5 cases) and control group (13 cases) was statistically significant (P<0.05); the incidence rate ratio difference cerebrospinal fluid leakage in study group (2 cases) and control group (8 cases) was statistically significant (P<0.05); the placed time of the study group was more than 10 d (42%) and control group (24%) the number of cases, the difference was statistically significant (P<0.05). Conclusion: In situ bone chip replantation is effective in preventing intracranial infection after external ventricular drainage. 2017 Vol. 47 (12): 914-915,918 [Abstract] ( 596 ) HTML (1 KB)  PDF (2659 KB)  ( 688 ) 916 Clinical analysis of 674 cases of foreign bodies aspiration in the respiratory tract in children ZHANG Hui, HUANG Qiuling, ZHANG Weixi, LI Changchong. Objective: To investigate the clinical characteristics of foreign bodies aspiration in the respiratorytract in children. Methods: A retrospective study was carried out, which involved 674 children of foreign bodies aspiration, who received therapy in the Second Affiliated Hospital & Yuying Children’s Hospital of Wenzhou Medical University from January 2011 to December 2016. Results: The ages of the children ranged from 6-month to 12-year old, and the duration of symptoms ranged from 1 hours to 12 months prior to medical intervention. From the initial 674 involved cases, 586 (86.94%) were caused by vegetable foreign bodies, 36 (5.34%) were pieces of animal food, other 38 (5.64%) were special-type foreign bodies, and the remaining 14 (2.08%) were other types. In those 674 cases, 641 children (95.25%) required therapy of rigid bronchoscopy, 19 (2.82%) received fiberoptic bronchoscopy, 11 (1.63%) were treated by laryngoscopy, 2 received thoracotomy, and 1 coughed up foreign body by himself. As a result of the treatment, 672 recovered after medical intervention, 1 was left with serious neurological sequelae and 1 died. Conclusion: Respiratory tract foreign body aspiration in children leads to severe adverse events. In order to diagnose and treat the children with foreign bodies timely, it is crucial to perform a comprehensive early examination for the children with foreign bodies. Rigid bronchoscopy is one of the most popular methods being used in clinical in removing respiratory airway foreign bodies, however, the advantages of fiberoptic bronchoscopy also should not be neglected. 2017 Vol. 47 (12): 916-918 [Abstract] ( 613 ) HTML (1 KB)  PDF (1467 KB)  ( 813 )

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