1.Mechanisms of lung injuries exposed to sulfur mustard:recent advances
Xiaoji ZHU ; Shuangshuang ZHU ; Yuxu ZHONG
Chinese Journal of Pharmacology and Toxicology 2015;(5):859-864
Sulphur mustard (SM) is a corrosive alkylating agent that is likely to be absorbed in vivo through the lungs,eyes and skin into internal organs. SM not only can produce its peculiar cytotoxic?ity thought to be mediated by the alkylation of DNA,protein and nucleic acids,but is a strong mutagen and carcinogen. However,whatever the way SM poisoning occurs,lungs are the most vulnerable, and early death is mainly carused by both the acute respiratory distress syndrome and pulmonary infec?tion. In this review,we analyzed SM-induced lung injury mechanisms.
2.Construction and identification of KIF18A protein baculovirus expression system
Zedong ZHAO ; Chenghua FU ; Shuangshuang LIANG ; Changjun ZHU ; Lina PAN
International Journal of Biomedical Engineering 2015;38(2):73-76,后插4
Objective KIF18A is a protein that has close relation with mitotic regulation and tumor development.This study aimed to establish KIF18A protein expression system in baculovirus,which may help to realize high efficient synthesis of KIF1 8A protein in vitro.Methods Transfer vector was constructed with molecular cloning method,and recombinant baculovirus were obtained through gene transposition.KIF18A protein was expressed by transforming recombinant baculovirus into infected insect Sf-9 cells to realize the synthesis efficiency.Results It was confirmed by DNA sequencing,microscopic observation and Western Blot that the KIF18A protein expression system in baculovirus was successfully established.Conclusions Conditions for transfer vector and recombinant virus transfection are defined,and high efficient KIF18A protein baculovirus expression system are successfully constructed.
3.Effect of cocaine-amphetamine-regulated transcript peptide on the content of 4-hydroxy-2-noneral and infarct volume after cerebral ischenia/reperfusion in mice
Zhenhan ZHU ; Dujuan SHA ; Qiming LI ; Jin LI ; Yong HAN ; Shuangshuang GU ; Jun ZHANG
International Journal of Cerebrovascular Diseases 2012;20(2):142-146
Objective To investigate the effect of cocaine-amphetamine-regulated transcript peptide (CART) on the content of 4-hydroxy-2-noneral (HNE) and infarct volume after cerebral ischemia/reperfusion in mice.Methods A total of 96 healthy male mice were randomly divided into four groups:ischemia/reperfusion (n =27),CART (n =27),normal saline control (n =27) and sham operation (n =15) groups.A middle cerebral artery occlusion (MCAO) model was induced.Two hours after MCAO,CART 55-102 and equivalent normal saline were injected respectively via the tail veins of mice in the CART group and the normal saline control group,and then they were injected every other 24 hour.The neurological scores,infarct volume and the HNE content of lipid metabolism of oxidative stress were performed and detected respectively at 12,24,48 and 72hours after reperfusion.Results CART could significantly improve the neurological deficit scores (all P <0.05) and reduce infarct volume (all P<0.05) at different time points after ischemia/reperfusion.The content of HNE was upregulated (all P<0.05) at different points after referfusion.CART could significantly down-regulate the increased HNE levd in brain after ischemia (all P<0.05).Conclusions CART may protect ischemic brain injury in mice by inhibiting lipid peroxidation.
4.Surface electromyographic examination of the coordination of the spastic upper limb muscles of stroke survivors
Shuangshuang CHENG ; Xiaoping GAO ; Xiaofei ZHU ; Xu ZHANG ; Dong WANG ; Li FU ; Ying CHENG
Chinese Journal of Physical Medicine and Rehabilitation 2017;39(5):342-346
Objective To explore any changes in the surface electromyography (sEMG) signals measured on the spastic upper limb muscles of stroke parents during maximum isometric voluntary contraction and to analyze any abnormal synergy patterns quantitatively in order to design better rehabilitation programs for developing coordination.Methods Ten stroke survivors with hemiparesis were selected into a patient group and ten healthy counterparts were recruited into a control group.sEMG signals were recorded bilaterally from the flexor carpi ulnaris (FCU),biceps brachii (BB),triceps brachii (TB) and deltoid (D) during maximum isometric voluntary contractions involving wrist flexion and extension,elbow flexion and extension,and shoulder abduction.The two groups' co-contraction ratios (CR) and co-activation ratios were calculated and compared.Results During elbow flexion and extension the average CR of the BB on the affected side was significantly higher than that on the unaffected side and also significantly higher than the control group average.The average CR of the TB on the affected side was significantly higher than that of the healthy controls.In all cases the average CR of the BB was larger than that of the TB.The difference in CR between the TB and the BB on the affected side was significantly larger than on the unaffected side and the control group average.During elbow flexion,the co-activation ratio of the FCU,TB and D on the affected side was significantly higher than on the unaffected side and among the healthy controls,and the co-activation ratio of the FCU on the affected side was significantly higher than that of the D and TB.During elbow extension,the co-activation ratio of the FCU,BB and D on the affected side was significantly higher in the same way,and the co-activation ratio of the FCU on the affected side was again significantly higher than that of the D and BB.During wrist flexion,the average co-activation ratio of the BB and D on the affected side was significantly greater than that on the unaffected side and among the healthy controls,and the co-activation ratio of the BB on the affected side was significantly higher than that of the D and TB.During shoulder abduction,the co-activation ratio of the BB on the affected side was significantly larger than on the unaffected side and among the healthy controls.Conclusion After a stroke the upper limbs often show flexor spasticity and abnormal synergy patterns.Rehabilitation strategy should be adopted to tackle these so as to enhance overall limb coordination.
5.Promotion of proliferation of prostate in aged rats by low-dose di(2-ethylhexyl) phthalate
Dongyan HUANG ; Shuangshuang WU ; Jing ZHU ; Xiaoyan MAO ; Lei LI ; Jianhui WU ; Zuyue SUN
Chinese Journal of Pharmacology and Toxicology 2017;31(6):642-648
OBJECTIVE To investigate the proliferation effect of di(2-ethylhexyl) phthalate (DEHP) on prostate in aged rats at the environmental exposure dose and the possible mechanism.METHODS Thirty-two male Sprague-Dawley rats,aged 1.5 years,were randomly divided into 4 groups (8 rats per group) and treated with DEHP (30,90 and 270 μg· kg-1,ig) and vehicle once daily respectively for 4 weeks.All the animals were anesthetized with pentobarbital sodium and sacrificed on the day subsequent to the last treatment.① Abdominal aortic blood samples were collected,and serum estradiol (E2),testosterone (T) and prolactin (PRL) levels were assayed by ELISA.② The prostate tissues were dissected and categorized into different lobes,weighed and measured.The prostate relative mass was calculated.③ The morphological changes were detected by HE staining and prostate epithelial height was analyzed with microscopic image analysis software.RESULTS Compared with vehicle control group,the prostate relative mass,dorsolateral prostate mass,and dorsolateral prostate index in DEHP 270 μg· kg-1 group were significantly higher (P<0.05).The height of the ventral prostate epithelium in DEHP 30,90 and 270 μg· kg-1 groups was increased significantly (P<0.01),so was the height of dorsal prostate epithelium in DEHP 270 μg· kg-1 group (P<0.01).There were no significant changes in levels of E2,PRL or T in DEHP 30,90 and 270 μg· kg-1 groups,but the ratios of E2/T in DEHP 30 and 270 μμg· kg-1 groups were increased significantly (P<0.05).CONCLUSION Low-dose DEHP could promote the proliferation of prostatic hyperplasia in the aged rats,which might be associated with the relative levels of endogenous hormone.
6.An optimization technique to purify the pre-ribosome and ribosome from mammalian cells using continuous sucrose density gradient ultracentrifugation
Shuangshuang LIANG ; Meichao JI ; Xiaoqing HU ; Chenghua FU ; Changiun ZHU ; Zhixiong DONG
International Journal of Biomedical Engineering 2015;38(5):262-265,281
Objective To purify pre-ribosome and ribosome of mammalian ceils using continuous sucrose density gradient ultracentrifugation.Methods Continuous sucrose density gradient was established by ultracentrifugation,and the continuous sucrose density gradient of 10%-30% and 10%-45% were used to extract the pre-ribosome and ribosome in mammalian cells,respectively.The mammalian cell lysis buffer was added to the established continuous sucrose density gradient.Pre-ribosome and ribosome with different sedimentation coefficients were collected and the A260 absorbance of each sample was measured.Proteins of each sample were extracted to detect the large subunit protein,RPL15 by Western Blot.Results Large subunit ribosomal protein RPL15 exists on 60S of the pre-ribosome,and also on 60S,80S and polyribosome of mature ribosome.Conclusions The continuous sucrose density gradient,which is established by the swing-out rotor,can be used to isolate the pre-ribosome and ribosome of mammalian cells rapidly.This method has the advantages of good separation effect and simple operation,which provides a good method for rapid and large amount preparation and separation of various kinds of ribosomes.
7.Modulation effect of chromosome kinesin protein KIF4A on cisplatin resistance of A549/DDP cells
Chenghua FU ; Xiaoqing HU ; Shuangshuang LIANG ; Meichao JI ; Zhixiong DONG ; Changjun ZHU
International Journal of Biomedical Engineering 2015;38(6):327-330,335
Objective To investigate the process that chromosome kinesin KIF4A promote cisplatin resistance in lung cancer cells.Methods Reverse transcription PCR (RT-PCR) and Western Blot experiments were performed to analyze the expression of KIF4A in lung cancer cells A549 and cisplatin (DDP) resistant cells A549/ DDP.Cell transfection, RNA interference (RNAi) experiments and thiazolyl blue tetrazolium bromide (MTT) assays were carried out to examine cell proliferation of A549 cells with overexpression of exogenous KIF4A and A549/DDP cells with depletion of endogenous KIF4A after cisplatin treatment.Results Expression of KIF4A in A549/DDP cells was higher than that in A549 cells.With overexpression of exogenous KIF4A, A549 cells displayed drug resistance to cisplatin.On the contrary, depletion of endogenous KIF4A in A549/DDP cells resulted in cisplatin sensitivity.Conclusions Chromosome kinesin KIF4A involves in the regulation of cisplatin resistance in lung cancer cells and KIF4A may be a potential and effective new biological target for treatment of lung cancer cisplatin resistance.
8.Revision of the Theory of Mind Tests for Children with Autism Spectrum Disorder
Weiting SHAO ; Sheng XU ; Hua FENG ; Shaoju JIN ; Shuangshuang ZHU ; Dan LI ; Zhen ZHANG ; Qiao ZHOU ; Xianjin XU
Chinese Mental Health Journal 2017;31(2):144-149
Objective:To amendment a tool of theory of mind (TOM) tests that can be applied to children with autism spectrum disorder (ASD) in mainland China,and assess the ability of ToM of children with ASD.Methods:The items of the ToM tests were revised by observing and recording events of children's life.Totally 200 normal children were selected in line with the standard from kindergarten to Grade 6 for formal tes-ting.With 156 valid data,Pearson correlation coefficient and Cronbach α coefficient were established for the test.Three experts were invited to make sure the content validity.Researchers randomly selected 30 normal children for retest purpose after 2 months.Twenty five children with ASD were tested and compared with normal children's test scores.Results:The revised test included 39 entries,which was divided into three sub-tests,42 points in total.The correlation coefficient of three subtests of the tests was 0.54-0.77,the correlation coefficient between the test and the subtest was 0.62-0.93 (Ps < 0.01).The scores of three experts for the test were 114,108,and 105.The total scores and subtest scores were lower in children with ASD than in normal children (Ps <0.01).The Cronbach a coefficient of the test was 0.84,Cronbach a coefficients for three subtest were 0.83,0.80,and 0.78,respectively.The retest reliability was 0.84,and reliabilities for three subtest were 0.75,0.74,and 1.00.Conclusion:The revised Theory of Mind Tests for Children with Autism Spectrum Disorder are fulfilled mostly in line with psychometric testing requirement.It might be a selection to measure the ability of theory of mind of children with ASD in mainland China.
9.Measurement of corneal thickness by optical coherence tomography angiography
Peiwen ZHU ; Xuexiang ZOU ; Kangcheng LIU ; Yun HAN ; Zhirong LIN ; Lei YE ; Mei SHEN ; Honghua KANG ; Shuangshuang ZHOU ; Gang TAN ; Yi SHAO
Recent Advances in Ophthalmology 2017;37(8):732-735
Objective To analyze the thickness of cornea and corneal epithelium in healthy subjects by optical coherence tomography angiography (OCTA).Methods Totally 100 healthy subjects aged between 20 and 30 years were analyzed by OCTA technique.Using AngioVue OCTA system of retinal imaging mode,and using SSADA algorithm for imaging,the cornea and the corneal epithelium in the central corneal diameter range of 9 mm were measured.The differences of corneal and corneal epithelial thickness in different gender regions were compared.Results In the male and female group,the corneal central total thickness were (559.92 ±33.26) μm and(540.06 ±31.63)μm,and the corneal epithelial thickness were(57.78 ±4.88) μm and(56.88 ±4.57) μm,The total central corneal thickness and central corneal epithelial thickness of the male were greater than those of the female,the difference was statistically significant (t =3.06,2.10;all P < 0.05).The cornea of male was the thickest at S5,S7 and SN9,there were significant differences at S5 and S7 compared with female (t =2.93,2.83;all P < 0.05);The female cornea was the thickest at S5,SN7 and SN9,and the difference was significant at S5 compared with male.The cornea of male subjects was the thinnest at IT,which was statistically significant only at IT5 compared with female subjects in the same area (t =2.02,P < 0.05);The cornea of female subjects was the thinnest at T5,IT7 and T9,which was statistically significant only at T5 and T9 compared with male subjects in the same region (t =2.63,2.20;all P < 0.05);There was significant difference in corneal thickness between male and female at ST (t =3.1 1,2.79,2.33;all P < 0.05).The corneal epithelium was the thickest at IT5,I7,and I9,and the lowest at S5,S7 and S9,and there was no significant difference compared with female in the same region (all P > 0.05).The corneal epithelium of female at the IT5,T7,N9 were the thickest,SN5,S7,S9 were the thinnest;Except for M2 and SN5,there was no significant differences in corneal epithelium between male and female groups (all P > 0.05).Corneal central epithelium accounted for the largest percentage of total corneal thickness,and gradually decreased from inside to outside.Conclusion OCTA can be used to measure the thickness of corneal and corneal epithelial regions.
10.Effect of reduced alpha - cardiac actin 1 gene expression on embryonic cardiomyocytes H9C2 cell apoptosis of rats and its mechanism
Shuangshuang YUAN ; Dongfang SUN ; Meijun XIE ; Binlu ZHU ; Hong JIANG ; Hongkun JIANG
Chinese Journal of Applied Clinical Pediatrics 2018;33(1):58-61
Objective To explore the effect of reduced alpha - cardiac actin 1(ACTC1)gene expression on the rat embryonic cardiomyocytes H9C2 cell apoptosis and its mechanism. Methods The rat embryonic cardiomyocytes H9C2 cell was cultivated;the rat embryonic cardiomyocytes H9C2 cell was transfected with ACTC1 - small interfering RNA(siRNA),and at 24 h,48 h,72 h after transfection,the cells were collected for extraction and purification of RNA, the real - time quantitative PCR(qPCR)method was used to detect the expression level of ACTC1 gene;and the termi-nal deoxynucleotidyl transferase - mediated dUTP - biotin nick end labeling assay(TUNEL)method was used to ex-plore the effect of reduced ACTC1 gene expression on the rat embryonic cardiomyocytes H9C2 cell apoptosis. Western blot was used to detect the expression of Cyto C,cysteine - containing aspartate - specific proteases(Caspase)- 3, Caspase - 8,Caspase - 9,Bcl - 2 and Bax. Results The expression of ACTC1 mRNA detected by qPCR decreased compared with that of the scramble siRNA group in 24 h,48 h,72 h(0. 80 vs. 1. 00,0. 20 vs. 1. 00,0. 25 vs. 1. 00),and in the ACTC1 - siRNA group decreased significantly at 48 h,72 h,and the difference was statistically significant(t =4. 245,P < 0. 05);TUNEL positive cells rate significantly increased in the ACTC1 - siRNA group(80%)compared with that in the scramble siRNA group(20%),and the difference was statistically significant(P < 0. 05);Western blot also confirmed that the expression of Caspase - 3,Caspase - 9,Cyto C and Bax/ Bcl - 2 were accordingly increased (0. 91 ± 0. 12 vs. 0. 59 ± 0. 01,0. 48 ± 0. 09 vs. 0. 24 ± 0. 03,0. 92 ± 0. 03 vs. 0. 45 ± 0. 01,2. 25 ± 0. 26 vs. 1. 16 ± 0. 12),and the differences were statistically significant(t = 2. 821,7. 336,2. 420,0. 798,all P < 0. 05);but the expre-ssion of Caspase - 8 had no obvious change,and the difference was not statistically significant (P > 0. 05 ). Conclusions Reduced ACTC1 gene expression can induce the rat embryonic cardiomyocytes H9C2 cell apoptosis perhaps mainly through endogenous mitochondrial signal transduction pathways.