3.Homology modeling of mosquitocidal Cry30Ca2 of Bacillus thuringiensis and its molecular docking with N-acetylgalactosamine.
Xin Min ZHAO ; Pan Deng ZHOU ; Li Qui XIA
Biomedical and Environmental Sciences 2012;25(5):590-596
OBJECTIVETo investigate the theoretical model of the three-dimensional structure of mosquitocidal Cry30Ca2 and its molecular docking with N-acetylgalactosamine.
METHODSThe theoretical model of Cry30Ca2 was predicted by homology modeling on the structure of the Cry4Ba. Docking studies were performed to investigate the interaction of Cry30Ca2 with N-acetylgalactosamine on the putative receptor.
RESULTSCry30Ca2 toxin is a rather compact molecule composed of three distinct domains and has approximate overall dimensions of 95 by 75 by 60Å. Domain II is a helix bundle, Domain II consists of three antiparallel β-sheets, Domain III is composed of two β-sheets that adopt a β-sandwich fold. Residue 321Ile in loop1, residues 342Gln 343Thr and 345Gln in loop2, residue 393Tyr in loop3 of Cry30Ca2 are responsible for the interactions with GalNAc via 7 hydrogen bonds, 6 of them were related to the oxygen atoms of hydroxyls of the ligand, and one to the nitrogen of the ligand.
CONCLUSIONThe 3D structure of Cry30Ca2 resembles the previously reported Cry toxin structures but shows still some distinctions. Several residues in the loops of the apex of domain II are responsible for the interactions with N-acetylgalactosamine.
Acetylgalactosamine ; chemistry ; Amino Acid Sequence ; Animals ; Bacterial Proteins ; chemistry ; pharmacology ; Catalytic Domain ; Culicidae ; drug effects ; Endotoxins ; chemistry ; pharmacology ; Hemolysin Proteins ; chemistry ; pharmacology ; Insecticides ; chemistry ; pharmacology ; Models, Molecular ; Molecular Sequence Data ; Protein Conformation
4.Role of tumor necrosis factor like ligand-1A aberrance in the generation and differentiation of peripheral blood Th17 in rheumatoid arthritis
Min ZHOU ; Rui LIU ; Xia LI ; Lingyun SUN
Chinese Journal of Rheumatology 2013;17(8):518-521
Objective To investigate the role of TL1A in the generation and differentiation of peripheral blood Th17 in rheumatoid arthritis.Methods The peripheral blood mononuclear cells (PBMCs) of RA were isolated and stimulated with PHA in the presence or absence of TL1A.Naive CD4+ T cells from RA was cultured under Th17-polarizing conditions with or without TL1A.The percentage of Th17 was detected by flow cytometry (FCM) analysis.The DR3 expression on CD4+ T cells from PBMCs of RA patients and healthy controls (HC) were analyzed by using FCM.Data were analyzed with t test and U test.Results TL1A could significantly up-regulate the percentage of Th17 in PBMCs of RA [(7.5±2.3)% vs (5.2±1.5)%,t=2.647,P<0.05].Compared to the HC groups,TL1A could significantly induce Th17 differentiation from naive T cells [(37.7±1.9)% vs (29.5±2.0)%,t=6.455,P<0.05].The percentage of CD4+DR3+ T cell in PBMCs of RA [(0.56±0.87)%] was significantly higher than that of HC [(0.13±0.04)%,P<0.05].The percentage of CD4+DR3+ T cell in PBMCs when stimulated with PHA was increased in RA patients,[(4.51±1.34)% vs (1.11±0.29)%,t=2.915,P<0.05],but no obvious increase in HC [(0.199±0.104)% vs 0.072%±0.029)%,t=1.644,P=0.1988].Conclusion TL1A can promote the generation and differentiation of Th17 in the PBMCs of RA,and this effect may be mediated by the binding of TL1A with DR3.
5.Effect of aging on olfaction center function: a magnetic resonance imaging study
Ying LIANG ; Weining HUANG ; Min CHEN ; Nina ZHOU ; Xia GONG
Chinese Journal of Geriatrics 2014;33(1):39-43
Objective To investigate the localization,zone and activation intensity of olfactory center in young versus elderly healthy volunteers by functional magnetic resonance imaging (BOLD-fMRI),so as to elucidate the effect of age on olfactory center in healthy population.Methods Thirteen right-handed healthy adult volunteers were recruited and divided into two groups:young group (5 males and 3 females,mean aged 23 years) and elderly group (2 males and 3 females,mean aged 69.2 years).The olfactory stimulus was r-undecalactone,and it was given according to a block design.The fMRI detection was performed on Philips Achieva 3.0 T MR scanner,and data of BOLE-fMRI was processed and analyzed to get cerebration image by using SPM2.Results In groupaveraged maps,both young and elderly group showed significant olfactory activation in right parahippocampal gyrus,left hippocampal sulcus,right and left superior temporal gyrus,etc,subcortical activation in right thalamus,dorsal pons,and cerebellum activation in cerebellar vermis.Activations in right inferior frontal gyrus,right middle frontal gyrus,right medial occipito-temporal gyrus and right fimbria of hippocampus were observed only in young group,while activation in bilateral middle temporal gyrus was observed only in elderly group.Activation area was apparently smaller and activation degree was lower in elderly group than in young group.Activation intensity in right superior parietal lobule and bilateral superior temporal gyri was higher in male group than in female group (t=13.7,6.08,5.36,respectively,all P<0.001).Conclusions The intensity of activation in olfactory center is lower in the elderly than in the young,and absence of part of the active regions is found in the elderly,which demonstrates the regression of olfactory center in the elderly.The olfactory center shows right-predominant activation,and olfactory activation intensity in some cortical regions is higher in males than in females.
6.The expression of tight junction protein claudin-5 after spinal cord injury about rats
Min CUI ; Xuenong HE ; Changlong ZHOU ; Xiaohui XIA ; Guangwei ZHANG
Chinese Journal of Nervous and Mental Diseases 2014;45(2):79-82
Objective To examine the expression of tight junction protein claudin-5 in blood-spinal cord barrier (BSCB) after spinal cord injury about rat. Methods One hundred-twenty adult male SD rats were randomly divided into blank group (60) and injured group (60). The animal model of spinal cord injury was established using modified Allen method. The expression of claudin-5 in BSCB was examined at 6 h, 12 h, 1 d, 3 d, 5 d and 7 d (five rats per time point). Western blot and RT_PCR were used to detect protein and mRNA expression levels of claudin-5, respectively. Results The success rate of spinal cord injury molding was 81.7%. In injured group, EB content increased gradually over time, reached the peak at the third day(0.9435 ± 0.0813)μg/g and then reduced gradually (P<0.05), EB content was signifi-cantly higher in injured group than in blank group. Claudin-5 mRNA expression in injured group reduced gradually over time and reached the lowest point at the third day(2.871 ± 0.527)and then increased gradually(P<0.05). Claudin-5 mRNA expression was significantly lower in injured group than in blank group(P<0.05). Claudin-5 protein expression in injured group reduced gradually over time, reached the lowest at the third day(0.072 ±0.008)and then increased gradually (P<0.05). Claudin- 5 protein expression was significantly lower in injured group than in blank group(P<0.05). Con-clusions The alteration of claudin-5 expression after SCI may lead to the permeability of BSCB, which may in turn con-tribute to the secondary spinal cord injury.
7.The effects of methylprednisolone pretreatment on the claudin-5 expression after spinal cord injury in rats
Min CUI ; Changlong ZHOU ; Xuenong HE ; Xiaohui XIA ; Guangwei ZHANG
Chinese Journal of Nervous and Mental Diseases 2014;(12):741-745
Objective To study the effects of methylprednisolone on the permeability of blood-spinal cord barrier (BSCB) and claudin-5 expression after spinal cord injury in rats. Methods The rat model of spinal cord injury was estab?lished using modified Allen method. SD rats were randomly divided into sham-operated group, spinal cord injury group and methylprednisolone pretreatment group. The permeability of BSCB and expression of claudin–5 were assessed at 12 h, 1, 3, 5, and 7 d after the onset of spinal cord injury (five animals per each time point). RT-PCR and Western blot were used to detect the expression of claudin-5. Results The success rate of the model was 84.0%. EB content was sig?nificantly higher in spinal cord injury group than in sham-operated group at each time point (F value 27.732,P<0.05). EB content was lower in methylprednisolone pretreatment group than in spinal cord injury group (F value 48.149,P<0.05). The mRNA expression of claudin-5 was lower in spinal cord injury group than sham-operated group at each time point (F value 12.248,P<0.05). The mRNA expression of claudin–5 was higher in methylprednisolone pretreatment group than in spinal cord injury group at each time point (Fvalue 15.316,P<0.05). The protein expression of claudin-5 was lower in spinal cord injury group than in sham operated group at each time point (Fvalue 18.108,P<0.05). The pro?tein expression of Claudin-5 was higher in methylprednisolone pretreatment group than spinal cord injury group at each time point (F value 20.247,P<0.05). Conclusions Methylprednisolone improves permeability of BSCB after spinal cord injury probably through enhancing claudin-5 expression in rats.
was lower in spinal cord injury group than in sham operated group at each time point (Fvalue 18.108,P<0.05). The pro?tein expression of Claudin-5 was higher in methylprednisolone pretreatment group than spinal cord injury group at each time point (F value 20.247,P<0.05). Conclusions Methylprednisolone improves permeability of BSCB after spinal cord injury probably through enhancing claudin-5 expression in rats.
8.Correlative analysis of lactic acid level with late-onset intracranial hematoma after craniocerebral injury
Changlong ZHOU ; Xuenong HE ; Xiaohui XIA ; Guangwei ZHANG ; Min CUI
Chinese Journal of Trauma 2014;30(9):883-885
Objective To analyze changes in lactic acid level in patients with late-onset intracranial hematoma after craniocerebral injury and investigate their relativity.Methods Forty-eight patients with late-onset intracranial hematoma after craniocerebral injury treated in our hospital between May 2009 and December 2012 were enrolled as observation group.There were 32 males and 16 females.Moreover,50 cases checked up in our hospital during the same period were studied as health population controls,including 35 males and 15 females.Level of lactic acid was measured on admission,at the time of definite diagnosis as well as at days 7 and 14 after treatment and compared between groups.Results Level of lactic acid was (1.77 ±0.21) mmol/L in control group and (1.82 ± 0.25) mmol/L in observation group respectively on admission (t =1.070,P > 0.05) ; Level of lactic acid was (3.32 ± 0.89) mmol/L in observation group at the time of definite diagnosis,which increased to (3.74 ± 1.16) mmol/L at days 7 after treatment and decreased to (1.89 ±0.75) mmol/L at days 14 after treatment.When diagnosed and treated for 7 days,level of lactic acid differed significantly between the two groups (P < 0.05).Level of lactic acid related to craniocerebral injury at each time point,but higher correlation coefficient was observed at the time of definite diagnosis and 7 days after treatment with 0.986 and 0.989 respectively.Conclusion Level of lactic acid relates to late-onset intracranial hematoma after craniocerebral injury,which can be used as reference for progression of the disease.
9.Xpert MTB/RIF assay in rapid diagnosis of tuberculous lymphadenitis
Lin ZHOU ; Qiang XIA ; Wenjun XIE ; Yumei FAN ; Min ZHU
Chinese Journal of Clinical Infectious Diseases 2016;9(6):502-506
Objective To assess the application of Xpert MTB/RIF assay in rapid diagnosis of tuberculous lymphadenitis.Methods Lymph node samples were collected from 1 02 clinically diagnosed patients with lymph node tuberculosis and 65 patients with other lymph node diseases from Zhejiang Provincial Hospital of Traditional Chinese and Western Medicine during January 201 4 and February 201 5. Xpert MTB/RIF,pathological examination and mycobacterium tuberculosis culture were conducted in all specimens of two groups.Taking clinical comprehensive diagnosis as the gold standard,the diagnostic sensitivity,specificity,positive predictive value,negative predictive value and diagnosis efficacy of three detection methods were assessed.The sensitivity of Xpert MTB/RIF in determining rifampicin resistance was analyzed using drug susceptibility testing as gold standard.Results The mean detection time of Xpert MTB/RIF was (2.2 ±0.2)h.Among 1 02 patients,Xpert MTB/RIF achieved higher sensitivity (96.1 %, 98 /1 02 ) than pathological examination (76.5%,78 /1 02 ) and mycobacterium tuberculosis culture (33.3%,34 /1 02)(χ2 =1 6.558 and 87.91 9,both P <0.01 ).Among 65 patients with other lymph node diseases,the specificity of all three detection methods was 1 00%.The receiver-operating-characteristic curve analysis demonstrated that diagnostic performance of Xpert MTB/RIF was better than that of other two methods.In 8 patients resistant to rifampicin confirmed by drug susceptibility testing,Xpert MTB/RIF detected 6 resistant-resistant patients.Conclusion Xpert MTB/RIF shows higher sensitivity and specificity in diagnosis of lymph node tuberculosis with the advantages of easy and rapid performance.
10.Effect of silencingRab11 by RNAi on invasion and migration of cervical cancer cell lines HeLa/SiHa and its mechanism
Yanyan KAN ; Jianhua ZHANG ; Min ZHOU ; Longzhen ZHANG ; Xia WANG
China Oncology 2016;26(3):238-244
Background and purpose:The expression ofRab11 gene was increased incervical cancer cell and may be involved in the cellular malignant transformation. This study used the sequence-speciifc siRNA knocking down the expression of Rab11 gene and aimed to investigate its effect on invasion and migration of cervical cancer cell lines HeLa/SiHa and its mechanism.Methods:HeLa/SiHa cells were divided into 2 groups: non-speciifc siRNA group transfected with unrelated siRNA (Rab11-NC) and Rab11 siRNA group transfected with Rab11 siRNA (Rab11siRNA). Western blot was used to examine the Rab11 protein expression. Cell migration and invasion were detected by cell scratch and Transwell invasion assay. Western blot was used to further investigate the expression of Rac1, matrix metal-loproteinase 2 (MMP2) and MMP9 which were critical for regulating cell invasion. Moreover, immunolfuorescence was used to identify intracellular location of Rac1 in HeLa/SiHa cells.Results:The Rab11 siRNA inhibited expression of Rab11 gene (P<0.01). The invasion and migration capacities of HeLa/SiHa cells were markedly inhibited in Rab11siR-NA group (P<0.05). The expression of Rac1 signiifcantly decreased (P<0.01). The expression of MMP2 and MMP9 de-creased (P<0.05) as well. The recruitment of Rac1 to protruding edge signiifcantly decreased following down-regulation of Rab11.Conclusion:Down-regulatedRab11 expression could inhibit the expression of Rac1, MMP2 and MMP9, and alter the location of Rac1, leading to suppression of HeLa/SiHa cells migration and invasion.