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Journal of Jiangsu University(Medicine Edition)
 
2014 Vol.24 Issue.06
Published 2014-11-30
Article
1
2014 Vol. 24 (06): 1- [Abstract] ( 521 ) [HTML 1KB] [ PDF 343KB] ( 1129 )
461 Topical application of IL-10 inhibits inflammation and reduces neural degeneration of the retina after corneal penetrating injury in rats
HUANG Yan-1, Zhou-Yue-Peng-2, Xiao-Shou-Hua-1, Zhang-Zhi-Jian-3
Objective: To investigate the mechanisms of that IL-10 inhibits inflammation and reduces neural degeneration of the retina after corneal penetrating injury in rats. Methods: A total of 50 adult female rats were randomly divided into three groups: A, the normal control group(8 rats); B, the corneal penetrating injury group(21 rats); and C, the IL-10 treated group(21 rats ). B and C groups were subdivided into the subgroups suffered from corneal penetrating injury for 24 hours，48 hours and 72 hours, respectively. The animals of C group suffered from corneal penetrating injury and then were treated with topical application of IL-10.Two rats in each group were used to make tissue sections of eyeball. The sections were immunofluorescence stained with antibodies against IL-1β, IL-6, TNF-α, neurofilament protein-200(NF200), α-Ⅱ spectrin and m-calpain. The relative contents of activated m-calpain and the degradation products of NF-200 and α-Ⅱ spectrin of the retina tissues were detected by western blotting. Results: The immunofluorescence staining of IL-1β, IL-6, TNF-α and m-calpain in normal retina were very weak, and became stronger in the retina of corneal penetrating injury groups. After treated with IL-10, the intensity of immunofluorescence staining of IL-1β,IL-6, TNF-α and m-calpain reduced. The immunofluorescence staining of NF-200 and α-Ⅱ spectrin were weaker in the corneal penetrating injury group than those in the normal group; after treated with IL-10, the intensity of immunofluorescence staining of NF-200 and α-Ⅱ spectrin became stronger than those in the corneal penetrating injury group. Western blotting showed that the relative contents of activated mcalpain and the degradation products of NF-200 and α-Ⅱ spectrin of the retina tissues in IL-10 treated group were lower than those in the corneal penetrating injury group. Conclusion: The expressions of IL-1β, IL-6 and TNF-α were upregulated after corneal penetrating injury and then activate the m-calpain localized in retinal tissues. The activate m-calpian could degrade the cytoskeleton proteins NF-200 and α-Ⅱ spectrin leading to nerve degeneration of retina; treatment with topical application of IL-10 could reduce the expressions of IL-1β, IL-6 and TNF-α and inhibit the activity of m-calpian, which protect the animals from nerve degeneration of the retina after corneal penetrating injury.
2014 Vol. 24 (06): 461- [Abstract] ( 1252 ) [HTML 1KB] [ PDF 3959KB] ( 1750 )
466 Effect of cholesterol on ROS generation and cell apoptosis in  human umbilical vein endothelial cells
LU Zhe, Shu-Bo, Wu-Liu-Song, Qian-Min-Zhang
Objective: To explore the effect of cholesterol on reactive oxygen species (ROS) generation and cell apoptosis in human vascular endothelial cells. Methods: HUVECs were treated with different cholesterol concentrations (12.5，25，50，100 mg/L) for 24 hours and were stimulated with 50 mg/L cholesterol at different times (0,6,12,24,48 h).Flow cytometry detected ROS generation.HUVECs were treated with different cholesterol concentrations and Nac.HUVECs survival rate were examined by MTT colorimetric assay and apoptosis rate were examined by flow cytometry. Results: The results showed that the level of ROS will all be elevated when HUVECs were treated with 12.5, 25, 50, 100 mg/L cholesterol for 24 h respectively. Compared with the control group, there was a significant difference (P<0.01).It had a significant difference between each groups (P<0.01) in the range of 50 mg/L, and appeared dosedependent manners. HUVECs were processed by 50 mg/L cholesterol for 6, 12, 24 and 48 hours , the level of ROS generation were significantly higher than control group(P<0.01);within 24 h emerge significant difference between each groups (P<0.01), and appeared the timedependent manners.The results of MTT showed that 12.5,25,50,100 mg/L cholesterol could reduce the cell survival rate ,and adding NAC could enhance survival rate.After HUVECs were processed by 50 mg/L or 100 mg/L cholesterol for 24 h, apoptosis rate was higher than control group (P<0.01), and the apoptosis rate were decline by adding antioxidants NAC. Conclusion: Cholesterol can enhance the level of ROS in HUVECs,and increase the HUVECs apoptosis.  ［Key words］cholesterol; human vascular endothelial cells; reactive oxygen species; apoptosis
2014 Vol. 24 (06): 466- [Abstract] ( 1204 ) [HTML 1KB] [ PDF 3254KB] ( 1398 )
470 CT quantitative analysis reflects the degree of ischemic necrosis and the level of glucose metabolism of non-small cell lung cancer
XU Wei-1, Dan-Xiu-Hong-1, Hu-Hui-1, Chen-Ye-Rong-1, Chen-Jian-Hua-2, Ding-Guo-Wen-3, Fan-Yu-4, Wang-Ya-Fei-1
Objective: To evaluate the feasibility of CT quantitative analysis reflecting the degree of ischemic necrosis and glucose metabolism level of nonsmall cell lung cancer(NSCLC). Methods: Fifty-two cases with NSCLC were enrolled in the study, the ischemia necrosis CT quantitative value(INCTQ) for NSCLC tumor on early enhancement CT images before surgery were measured. Glucose transporter 1 (Glut 1) and carbonic anhydrase Ⅸ (CA Ⅸ) protein expression with paraffin embedded specimen after operation were detected by immunohistochemical staining. Results: The INCTQ values of expression of Glut1negative, grade 1 and grade 2 were 0.35 ± 0.23, 0.52 ± 0.55 and 1.55 ± 1.20, respectively (F=10.633, P=0.000); the INCTQ values of the Glut 1 and CA IX nonexpression group, single expression group and coexpression group were 0.32±0.25, 0.85±0.83 and 1.37±1.27, respectively (F=4.449, P=0.017). Conclusion: The early enhancement CT could be used to appraise the ischemia necrosis degree, and reflect the level of glucose metabolism of NSCLC.  ［Key words］nonsmall cell lung cancer; glucose transporter protein 1; carbonic anhydrase Ⅸ； Xray computed tomography; ischemia necrosis
2014 Vol. 24 (06): 470- [Abstract] ( 1832 ) [HTML 1KB] [ PDF 5481KB] ( 1630 )
474 Effect of H3 receptor agonist on eosinophils and eotaxin of allergic rhinitis in guinea pigs
DOU Yi, Yang-Xu-Dong, Huang-Qiu-Sheng
Objective: To explore the influence of histamine receptor 3 (H3R ) agonist, IMETIT, on eosinophils and one of eosinophilic chemokines (eotaxin) in allergic rhinitis(AR) model using guinea pigs.  Methods: Thirty guinea pigs were randomly divided into three groups: control group, sensitized group without treatment and sensitized group with drug treatment. The sensitized group with or without drug treatment were immunized by ovalbumin (OVA) to establish AR model and the control group by saline to contrast, then they were stimulated intranasal by ovalbumin (1%) respectively. The times of sneezing and scratching nose in 30 min were recorded. The control group and the sensitized group without treatment were given gastric gavage by saline. And the sensitized group with drug treatment were given gastric gavage by saline (10 mL/kg) with IMETIT (30 mg/kg). Then we investigated the eosinophil infiltration degrees and eosinophilc chemokines content in nasal septum mucosa using HE staining and immunohistochemistry technology. Results: There was significant difference between the control group and the sensitized group with or without drug treatment in the numbers of sneezing and the numbers of scratching nose(P=0.000) . There was no significant difference between the sensitized group with or without drug treatment in the means of eosinophil and eotaxin positive expression rate in nasal septum mucosa(P=0.581, 0.676) . Conclusion: The eosinophil infiltration and the level of eotaxin of nasal septum mucous could not be reduced by histamine H3 receptor agonist, IMETIT.  ［Key words］H3 receptor agonist; allergic rhinitis; eosinophil; eotaxin
2014 Vol. 24 (06): 474- [Abstract] ( 709 ) [HTML 1KB] [ PDF 3155KB] ( 1590 )
478 Effect of aloeelodin on nonalcoholic steatohepatitis cells model
DAI Na-1, Zhou-Pei-2, Zhang-Ying-Ying-2, Wang-Bing-Fang-1
Objective: To study the effect of aloeemodin(AE) on non-alcoholic steatohepatitis cell model. Methods: Thirty percent fat emulsion were added into the culture medium cultured normal human liver cell line L-02, then the cells were divided into control group and experiment groups in which culture medium containing 30% fat emulsion at different concentrations, which were 0.25ml/L, 0.5 mL/L, 0.75 ml/L, 1.0 ml/L, respectively. After 72 hours culture, the cells were stained by oil red O, then observed the morphology and the intracellular lipid droplets; and mesured the alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG) contents in liquid culture; choosed the most appropriate concentration of 30% fat emulsion(0.5 mL/L FA group) to establish the nonalcoholic steatohepatitis model. Then the cells were randomized into the control group, model group and experimental groups in which culture medium the concentrations of aloe emodin were 10-4 mmol/L, 10-5 mmol/L, 10-6 mmol/L, respectively. The number of intracellular lipid droplets, triglyceride content and indicators of ALT, AST, superoxide dismutase (SOD), malonaldehyde (MDA), endotoxin, tumor necrosis factor-α (TNF-α) in culture medium were determined. Results: The model of NASH liver cells were successfully established. Three groups were added with different concentration of AE intervention, after 72 hours, their ALT, AST, MDA, endotoxin, TNF-α in liquid culture decreased, SOD content increased compared with the model group (P<0.05). Conclusion: AE could improve liver cell lipid peroxidation degree, and reduce inflammation and cell damage, which correlated positively with the time and concentration, within a given range. ［Key words］nonalcoholic steatohepatitis; aloeelodin; cell model; cell injury
2014 Vol. 24 (06): 478- [Abstract] ( 1469 ) [HTML 1KB] [ PDF 4058KB] ( 1432 )
483 Preparation of 3-D ordered macroporous silica as drug carrier and release characteristics in vitro
WANG Liang, Li-Qian-Qian, Yu-Jiang-南, Xu-Xi-Ming
Objective: To prepare and characterize 3D ordered macroporous (3DOM) silica as drug carrier for poorly water soluble drugs and investigate its in vitro release characteristics. Methods: 3DOM silica carrier was prepared using PMMA colloidal crystal template method and drug was incorporated into pores of 3DOM silica matrix through a solvent deposition procedure. The pore structure was characterized was studied by scanning electron microscopy (SEM). The solid state of drug in 3DOM silica was evaluated by X-ray diffraction (XRD). Inter-molecular interaction of drug and matrix was obtained using a FT-IR spectrometer. The release property in vitro was investigated. Results: Silica carrier obtained showed exactly a three-dimensional ordered macroporous structure. There was a significant loss of the drug crystallinity at drug-carrier ratio of 1 ∶3 and 1 ∶5, and the release rate in vitro reached 80% in 2 hours. Conclusion: 3DOM silica was successfully prepared using colloidal crystal template method, and the dissolution of poorly water soluble drug was improved significantly.  ［Key words］threedimensional ordered macroporous silica; carriers; poorly water soluble drugs; in vitro dissolution
2014 Vol. 24 (06): 483- [Abstract] ( 1069 ) [HTML 1KB] [ PDF 3509KB] ( 1898 )
487 Construction and identification of cagQ gene-deleted mutant in type Ⅳ secretion system of H.pylori
YAO Yi-Zheng, Wang-Hua, Ni-Ying, Shen-Yi-Xin, Xu-Chi, Sun-Feng-Ying, Shao-Shi-He
Objective: To construct the cagQ gene-deleted mutant of Helicobacter pylori (H.pylori) Methods: Polymerase chain reaction (PCR) was used to amplify the homologous gene fragments (upstream and downstream of the openreading frames) of cagQ gene. The homologous gene fragments were connected to pMD18-T vector and the fragments were cut down by restriction enzyme. Then the homologous fragments were inserted into the pBluescript SK II(-) which includes kanamycin resistance gene. The suicide plasmid pBlueKM40-△cagQ was transferred into H.pylori through electroporation. The complete recombinant strain grown on agar plates supplemented with kanamycin was selected by PCR and confirmed through sequences analysis. Results: The suicide plasmid pBlueKM40-△cagQ was identified by restriction enzyme digestion and was transferred into H.pylori. The result of the genedeleted mutant was confirmed through PCR and sequencing. Conclusion: We generated the cagQ gene-deleted mutant of H.pylori(Hp26695-△cagQ).  ［Key words］Helicobacter pylori; type Ⅳ secretion system; cagQ; gene-deleted mutant
2014 Vol. 24 (06): 487- [Abstract] ( 1305 ) [HTML 1KB] [ PDF 3744KB] ( 2342 )
491 Establishment of the onestep DNA extraction method of human papillomavirus in cervical epithelium samples
ZHAO Qi
Objective: To develop an one-step method on DNA extraction of human papillomavirus in cervical epithelium samples. Methods: The extraction of HPV-DNA in 200 cases collected from cervical epithelium samples was treated by column method of DNA extraction kit or one-step method. DNA purity and concentration of the samples were measured by the trace nucleic acid spectrophotometer. HPV genotypes were detected by PCR reverse dot blot(PCR-RDB) method. Results: DNA purity extracted by column method and onestep were (1.91±0.14) μg/mL and (1.86±0.19) μg/mL, respectively, and concentration of the HPV-DNA were (139.78±18.21) μg/mL and (124.36±17.35) μg/mL, respectively. DNA purity and concentration extracted by two methods were all not statistically significant(P<0.01). The rates of positive，single infection and multiple infection of HPV genotyping treated by the column method and onestep method were 17.5%(35/200), 7.5%(15/200), 10%(20/200) and 17.5%(35/200), 8%(16/200), 9.5%(19/200), respectively. The value of Kappa was 0.886， which indicated that there was a high degree of consistency. Conclusion: The one-step method on HPV DNA extraction was simpler, the DNA quality was high and suitable for HPV DNA genotyping of the cervical epithelium samples.  ［Key words］polymerase chain reaction; reverse dot blot ;human papillomavirus; DNA extraction; onestep method; separation method
2014 Vol. 24 (06): 491- [Abstract] ( 1083 ) [HTML 1KB] [ PDF 2896KB] ( 1844 )
495
ZHANG Shu-Ying-1, 2 , Lu-Xiao-Dong-1*, Wang-Yu-Yue-2
2014 Vol. 24 (06): 495- [Abstract] ( 1001 ) [HTML 1KB] [ PDF 2156KB] ( 1872 )
497 Effects of sitagliptin on oxidative stress and endothelin-1 in diabetic patients with carotid atherosclerosis
FENG Ya-Min-1, Lu-Yi-Bing-1, Miao-Hang-1, Jin-Jun-Fei-2, Zhu-Qun-1
Objective: To investigate the effects of sitagliptin on oxidative stress and endothelin-1(ET-1) in type 2 diabetic patients with carotid atherosclerosis. Methods: Sixty-two type 2 diabetic patients with carotid atherosclerosis were randomly divided into sitagliptin group(n=32) and control group (n=30). Patients in sitagliptin group taken sitagliptin 100 mg per day with their currently hypoglycemic drug treatment(dosage of insulin reduced) and patients in control group continued their hypoglycemic drug treatment. Malonaldehyde (MDA), the marker of oxidative stress, superoxide dismutase (SOD) and endothelin-1(ET-1) from the two groups (pre- and post-treatment) were determined and compared. Results: There were no significant differences in the levels of ET-1, MDA and SOD between two groups before treatment. After treatment, the levels of ET-1 and MDA in sitagliptin group were significantly decreased (P<0.01), and the activity of SOD was significantly increased (P<0.01). There were no significant differences in the levels of ET-1, MDA and SOD in control group between pretreatment and post-treatment (P>0.05). Conclusion: Sitagliptin has good effects on decreasing levels of oxidative stress and ET-1, which might protect from the diabetic macrovascular complications.  ［Key words］type 2 diabetes; carotid atherosclerosis; oxidative stress; sitagliptin
2014 Vol. 24 (06): 497- [Abstract] ( 1468 ) [HTML 1KB] [ PDF 2868KB] ( 1797 )
501 Analysis of serum high molecular weight adiponectin levels and  related factors in patients with essential hypertension
JU Chang-Nian-1, Qian-Wei-Yun-2, Zhu-Tian-Yi-2, et al
Objective: To detect serum high-molecular-weight adiponectin(HMW APN) level in hypertension patients and investigate the relationships between serum HMW APN level and blood pressure,obesity,glucolipid metabolism,insulin resistance and highly sensitive C-reactive protein(hs-CRP). Methods: A total of 76 subjects were enrolled in the study and divided into two groups by the definition of hypertension: the hypertension group(n=38) and the normal control group(n=38).Oral glucose tolerance test(OGTT) were performed in all subjects.Fasting plasma glucose(FPG), 2 h postprandial plasma glucose(2hPG), fasting insulin(FINS), 2 h postprandial serum insulin(2hINS) and lipids were also detected.Insulin resistance was assessed by homeostasis model assessment(HOMA-IR). Hs-CRP level was determined by chemiluminescence immunoassay and fasting serum HMW APN level was determined by ELISA. Results: The serum HMW APN level in hypertension group was significantly lower than that in normal control group [0.82(0.46-1.46) vs 2.53(1.70-5.75)ng/L,P＜0.01]. Pearson correlation analysis showed that serum HMW APN level was negatively correlated with SBP, DBP, MAP, BMI, WHR,TG,FPG,2 hPG, HOMAIR and hs-CRP(P＜0.05 or P＜0.01), and was positively correlated with HDL-C(P＜0.01). Multiple linear regression analysis showed that SBP and TG were independently related to serum HMW adiponectin level(P<0.05). Logistic regression analysis showed that HMW adiponectin was the independent protective factor for hypertension(P<0.01). Conclusion: Serum HMW adiponectin level was decreased in subjects with hypertension. Serum HMW adiponectin level was closely correlated with blood pressure, obesity, glucolipid metabolism,insulin resistance and hs-CRP.It suggests that HMW APN decline may play an important role in the pathogenesis of hypertension.  ［Key words］hypertension; high molecular weight adiponectin; obesity; insulin resistance
2014 Vol. 24 (06): 501- [Abstract] ( 1148 ) [HTML 1KB] [ PDF 3558KB] ( 1833 )
505 Expression of certain mitochondrion enzymes in cartilages of osteoarthritis
QIAN Wei-1, Bipin Kumar Rai1, Yuan-Ji-Shan-2, Liu-Yan-Fang-3, et al
Objective: To compare the expressions of seven important mitochondrion enzymes in osteoarthritis (OA) cartilages and normal cartilages. Methods: Specimens of human articular cartilage were collected from a total of 9 osteoarthritis patients and 12 normal controls . H&E staining showed the histologic structure of cartilage. Expressions of NDUFA1(NADH dehydrogenase ubiquinone 1 alpha subcomplex 1）, succinate dehydrogenase complex(SDHB), cytochrome b(Cytb), COX(cytochrome c oxidase), V-ATPase H(vacuolar-type H+-ATPase), cytosolic malate dehydrogenase (MDH), hydroxyacyl-coenzyme A dehydrogenase(HADHSC) were detected by RQ-PCR and Western-blot. Results: Compared with normal cartilage, synoviocytes proliferation and lymphocytes infiltration could be observed in osteoarthritis group. The expressions of NDUFA1 and COX increased in osteoarthritis cartilages, expressions of SDHB, Cytb, V-ATPase H decreased in osteoarthritis cartilages. The differences were statistically significant(P<0.05). There was no statistical significance in MDH and HADHSC expression between the two groups. Conclusion: Changes of some important mitochondrion enzymes may play an important role in pathopoiesis of osteoarthritis.  ［Key words］osteoarthritis; cartilage; mitochondrion; enzymes
2014 Vol. 24 (06): 505- [Abstract] ( 1350 ) [HTML 1KB] [ PDF 3723KB] ( 1834 )
510
QIANG Li, YAN Yong-Dong
2014 Vol. 24 (06): 510- [Abstract] ( 887 ) [HTML 1KB] [ PDF 2147KB] ( 1435 )
513
SHAO Jia-1, He-Ai-Qin-1*, Zhang-Yu-Quan-2, Liu-Ji-Bin-3
2014 Vol. 24 (06): 513- [Abstract] ( 1316 ) [HTML 1KB] [ PDF 3578KB] ( 1760 )
517
YUAN Ju-Fang, Chen-Zheng, Shao-Dong-Hua-*
2014 Vol. 24 (06): 517- [Abstract] ( 811 ) [HTML 1KB] [ PDF 2154KB] ( 1490 )
520
WANG Li, Lian-Yun, Zhao-Xiang-Dong, Li-Hao, Zhu-Yang-Quan-*
2014 Vol. 24 (06): 520- [Abstract] ( 642 ) [HTML 1KB] [ PDF 2197KB] ( 1662 )
522
HU Fei-Ya
2014 Vol. 24 (06): 522- [Abstract] ( 719 ) [HTML 1KB] [ PDF 2352KB] ( 1950 )
525
WANG Hong-Zhen-1, SHI Hong-Jin-2
2014 Vol. 24 (06): 525- [Abstract] ( 837 ) [HTML 1KB] [ PDF 2858KB] ( 3386 )
529
ZHANG Dong-Mei-1, Qian-Xiao-Qin-1, Sun-Xiao-Ning-1, Dan-Xiu-Hong-2*
2014 Vol. 24 (06): 529- [Abstract] ( 1174 ) [HTML 1KB] [ PDF 2862KB] ( 1365 )
532
GUO Yan-Fen, Han-Chao-*, Liu-Qian, Jiang-Wen-Jie, Gu-Da-Min, Ge-Zhi-Jun
2014 Vol. 24 (06): 532- [Abstract] ( 679 ) [HTML 1KB] [ PDF 2009KB] ( 1445 )
535
MENG Wen-Jun, Xu-Xin, Dai-Qiu-Xin, Yang-Jian-Gang, Qin-Ru-Juan, Chen-Wei-Feng
2014 Vol. 24 (06): 535- [Abstract] ( 864 ) [HTML 1KB] [ PDF 2142KB] ( 2438 )
538
TAO Zhi-Qiang-1, Song-Jie-2*, Xu-Biao-2, Huang-Wei-2, Wang-Lian-2, Zhang-Jing-Mei-2, Xie-Jun-2
2014 Vol. 24 (06): 538- [Abstract] ( 1112 ) [HTML 1KB] [ PDF 2144KB] ( 1946 )
541
WANG Fei-Fei-1, Shi-Chang-Xi-2
2014 Vol. 24 (06): 541- [Abstract] ( 870 ) [HTML 1KB] [ PDF 1472KB] ( 1623 )
543
LIU Yuan, Shao-Ji
2014 Vol. 24 (06): 543- [Abstract] ( 616 ) [HTML 1KB] [ PDF 1781KB] ( 1653 )
545
YI Li-Xian-1, 2 , Tang-Xin-Yi-1, Wang-Sheng-Jun-1*
2014 Vol. 24 (06): 545- [Abstract] ( 708 ) [HTML 1KB] [ PDF 2718KB] ( 2171 )
201406
BIAN Ji-Bu
2014 Vol. 24 (06): 201406- [Abstract] ( 595 ) [HTML 1KB] [ PDF 442KB] ( 1012 )
江苏大学学报:医学版
　

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