Objective:To observe the effect of electroacupuncture(EA)combined with oriented conductive bio-protein hydrogel(OCBH)on the recovery of nerve function in rats with complete spinal cord injury(SCI)and to explore its effect and mechanism on the formation and changes of glial scars.Methods:A total of 72 female Sprague-Dawley rats were randomly divided into groups according to the treatment received.A rat model of complete SCI was constructed using a spinal cord transection.Behavioral assessments,hematoxylin-eosin(H&E)staining,immunofluorescence staining,and Western blotting were performed at a fixed period after the operation.Results:The material group and the material+EA group obtained better results in the behavioral as-sessments(all P＜.05)and the H&E staining.In the immunofluorescence staining and Western blotting,the GFAP protein was expressed more and denser in the material group and the material+EA group than in the model group,and the density of the GFAP expression in the material+EA group was lower at week 12 than in the material group(all P＜.05).The expression of complement C3 in the model,material,and material+EA groups decreased in turn.Some inflammatory factors and the NF-kB signaling pathway showed similar results in the Western blotting(all P＜.05).The expression of the GDNF protein in the material+EA group was significantly higher than that in the model group and the material group(both P＜.01).Conclusion:EA combined with OCBH can promote the recovery of motor functions after SCI by facili-tating the formation of glial scars in the early stage,preventing the further spread of an inflammatory response that would affect the activation of A1/A2 astrocytes and change the morphology of glial scars at the spinal cord-material interface in its late stage.

Objective:To screen the biomarkers in the exhaled breath of mice exposed to benzene by using exhaled breath online analysis system.Methods:Thirty 8-week-old male C57BL/6 mice were randomly divided into six groups (0, 3, 32, 324, 648, and 1 296 mg/m 3) and treated with benzene vapour for 28 days. At the end of the exposure, the peripheral blood cell counts and blood glutathione (GSH) were detected. The content of malondialdehyde (MDA) in HL60 cells treated by mice plasma was examined. Exhaled breath data from mice were collected by Secondary electrospray ionization source high resolution mass spectrometry (SESI-HRMS). Targeted analysis underlying benzene metabolites and oxidative stress metabolites was performed to screen the biomarkers in exhaled breath. Results:After benzene exposure, the number of peripheral blood cells was decreased in different degrees, particularly in the white blood cells (WBC) number. The WBC in 32 and 324 mg/m 3 groups was declined by 27.76% and 52.87%, respectively compared to that in control group ( P<0.05). Meanwhile, compared with the control group, the GSH content of peripheral blood cells from 324 mg/m 3 group decreased by 13.16% ( P<0.05). In addition, MDA content was increased by 18.11% in HL60 cells treated with plasma from 324 mg/m 3 group mice ( P<0.05). The phenol, hydroquinone/catechol, benzenetriol and trans, trans-Muconic acid ( t,t-MA) in the exhaled gas of mice could be used as biomarkers for benzene exposure ( R 2>0.8, P<0.001). The peak intensity of five small molecular metabolites related to oxidative stress (ω-carboxylic fatty acid C 5H 10O 3, ω-carboxylic fatty acid C 6H 12O 3, glutamate, cysteine and MDA) increased with the increase of benzene concentration ( P<0.05), which was negatively correlated with WBC decline ( P<0.001), suggesting that these molecules mignt be used as biomarkers of benzene-induced toxicity. Conclusions:Phenol, hydroquinone/catechol, benzenetriol and trans, trans-Muconic acid ( t,t-MA) in exhaled breath of mice could be used as biomarkers for benzene exposure; ω-carboxylic fatty acid C 5H 10O 3, ω-carboxylic fatty acid C 6H 12O 3, glutamate, cysteine and MDA might be used as markers of benzene-induced toxicity.

Objective:To screen the biomarkers in the exhaled breath of mice exposed to benzene by using exhaled breath online analysis system.Methods:Thirty 8-week-old male C57BL/6 mice were randomly divided into six groups (0, 3, 32, 324, 648, and 1 296 mg/m 3) and treated with benzene vapour for 28 days. At the end of the exposure, the peripheral blood cell counts and blood glutathione (GSH) were detected. The content of malondialdehyde (MDA) in HL60 cells treated by mice plasma was examined. Exhaled breath data from mice were collected by Secondary electrospray ionization source high resolution mass spectrometry (SESI-HRMS). Targeted analysis underlying benzene metabolites and oxidative stress metabolites was performed to screen the biomarkers in exhaled breath. Results:After benzene exposure, the number of peripheral blood cells was decreased in different degrees, particularly in the white blood cells (WBC) number. The WBC in 32 and 324 mg/m 3 groups was declined by 27.76% and 52.87%, respectively compared to that in control group ( P<0.05). Meanwhile, compared with the control group, the GSH content of peripheral blood cells from 324 mg/m 3 group decreased by 13.16% ( P<0.05). In addition, MDA content was increased by 18.11% in HL60 cells treated with plasma from 324 mg/m 3 group mice ( P<0.05). The phenol, hydroquinone/catechol, benzenetriol and trans, trans-Muconic acid ( t,t-MA) in the exhaled gas of mice could be used as biomarkers for benzene exposure ( R 2>0.8, P<0.001). The peak intensity of five small molecular metabolites related to oxidative stress (ω-carboxylic fatty acid C 5H 10O 3, ω-carboxylic fatty acid C 6H 12O 3, glutamate, cysteine and MDA) increased with the increase of benzene concentration ( P<0.05), which was negatively correlated with WBC decline ( P<0.001), suggesting that these molecules mignt be used as biomarkers of benzene-induced toxicity. Conclusions:Phenol, hydroquinone/catechol, benzenetriol and trans, trans-Muconic acid ( t,t-MA) in exhaled breath of mice could be used as biomarkers for benzene exposure; ω-carboxylic fatty acid C 5H 10O 3, ω-carboxylic fatty acid C 6H 12O 3, glutamate, cysteine and MDA might be used as markers of benzene-induced toxicity.

Objective To investigate the accuracy and safety of using normal saline as a medium to guide the catheter lumen to assist the localization of the catheter tip. Methods This study included the patients with hematological malignancies in our fully implantable venous port. We enrolled 105 patients from January 2014 to December 2015 as control group, and 220 patients from January 2016 to June 2017 as the experimental group. The control group used the traditional fully implantable venous port after the chest X slice to determine the location of the catheter tip. The experimental group used the intracavitary electrocardiogram location technology to assist the complete implantable venous port catheter tip positioning catheter, then chest X film. Results No catheterization occurred in all the patients. The rate of catheter placement was 100％. In the experimental group, 179 patients (81.36％) had the best placement of catheter tip (i.e., superior vena cava right atrium junction, CAJ), and 35 patients (52.38％) in the control group had the best placement of catheter tip. The proportion of the catheter tip located in the best position (the superior vena cava auricular commissure and CAJ) in the experimental group was higher than that in the control group (χ2=29.615, P < 0.05). Conclusions By the injection of saline guided endocardial mapping real-time monitoring of totally implantable venous transfusion port catheter tip position, can guide the surgeon to grasp the real-time operational direction of totally implantable venous transfusion port catheter tip, accurately positioning the catheter tip position, improve the surgical success rate, worthy of clinical use.

Objective Based on the three-dimensional quality structure model,to construct the nursing quality evaluation index system for acute leukemia,in order to provide references for clinical evaluation of quality of nursing care for acute leukemia.Methods Based on the theory of Donabedian's structure-process-outcome quality structure model,through literature search,semi-structured interview,expert meeting,expert consultation and analytic hierarchy process,the quality evaluation index system and index weight for acute leukemia were determined.Results After two rounds of expert consultation,questionnaire response rates were 94.74％,100％;expert authority coefficients were 0.848,0.854;Kendall coordination coefficients were 0.273,0.420,and P values were all less than 0.01.The final index system consisted of 3 first-level indicators,8 second-level indicators,and 24 third-level indicators (3 structure indicators,7 process indicators,14 outcome indicators).Each of third-level indicators contained index names,calculation methods,and data collection methods.Conclusion The process of construction of nursing quality evaluation index system for acute leukemia is scientific,the contents are reasonable,which can reflect nursing characteristics of acute leukemia.

Objective: To establish the immortalized mouse brain microvascular pericytes model and to apply to the cerebrovascular toxicants screening study. Methods: Brain pericytes were isolated from 3 weeks of mice by tissue digestion. Immortalized pericyte cell line was constructed by infecting with LT retrovirus. Monoclone was selected to purify the immortalized pericyte cell line. The pericyte characteristics and purity were explored by immunocytochemistry. Cell proliferation was measured by using the Pomega MTS cell Proliferation Colorimetric Assay Kit. Pericytes were treated with 0, 160, 320, 640, 1 280, 2 560 μmol/L lead acetate, 0, 5, 10, 20, 40, 80 μmol/L cadmium chloride and 0, 5, 10, 20, 40, 80 μmol/L sodium arsenite in 24 hours. Cell toxicity of each group was determined by MTS assay, median lethal dose (LD₅₀) was calculated in linear regression. Results: Mouse brain pericytes were successfully isolated by tissue separation and enzyme digestion method. After immortalized by LT retroviruses, monoclone was selected and expanded to establish pericyte cell line. The brain pericytes exhibited typical long spindle morphology and positive staining for α-SMA and Vimentin. The proliferation of brain pericytes cell lines was very slowly, and the doubling time was about 48 hours. The proliferation of immortalized brain pericytes cell lines was very quickly, and the doubling time was about 24 hours. After lead acetate, cadmium chloride and sodium arsenite treatment for 24 hours respectively, gradual declines in cell viability were observed. The LD₅₀ of lead acetate was 2 025.0 μmol/L, the LD₅₀ of cadmium chloride was 36.6 μmol/L, and the LD₅₀ of sodium arsenite was 33.2 μmol/L. Conclusion The immortalized mouse brain microvascular pericyte model is established successfully by infecting with LT retrovirus, and can be applied to screen cerebrovascular toxicants. The toxicity of these toxicants to immortalized mouse brain microvascular pericyte is in sequence: sodium arsenite,cadmium chloride, lead acetate.

Objective: To investigate the effects of cerebral cavernous malformation 3 (CCM3) gene knockout on the lead exposure-induced blood-brain barrier malfunction in mice brain, and the relationship between CCM3 knockout and the Alzheimer's disease (AD). Methods: Wide type (WT) mice and CCM3⁺/- mice were divided into 4 groups, control group and lead exposed group in WT as well as CCM3^+/- mice. Lead exposed groups were treated with 0.05% lead acetate in drinking water for 12 weeks, while control group drink deionized water freely. Blood lead and brain lead levels in each group were detected by graphite furnace atomic absorption spectrometry. The brain tissue of each group was made into paraffin sections, whose morphology were observed by HE staining. The expression of Aβ_1-42 in brain tissue was detected by immunohistochemistry and the brain capillaries were labeled by VRGFR2. The protein expression of Claudin-5, ZO-1, and p-Tau was detected by Western blot. The brain tissue RNA was extracted and the relative expression of LRP-1 mRNA was detected by qRT-PCR. Results: The levels of blood lead WT (216.07±84.16) and CCM3^+/- (189.64±101.86) μg/L in lead exposed group were higher than those in control group WT (19.52±11.46) and CCM3^+/- (11.79±8.20) μg/L, the difference was statistically significant (t=4.18, P=0.006; t=3.79, P=0.016). The levels of brain lead WT (1.78±0.69) and CCM3^+/- (1.74±0.66) μg/L were higher than those in control group WT (1.06±0.87) and CCM3^+/- (0.97±0.64) μg/L, the difference was statistically significant (t=3.67, P=0.018; t=3.88, P=0.015). The HE staining showed no obvious lesions in the brain of each group of mice. The results of immunohistochemistry showed that there was no Aβ₁-42 deposition in the brain of mice in each group. The numbers of microvessels in the brain of CCM3^+/- mice in the lead exposed group were decreased. Compared with the relative expression levels of Claudin-5 (WT: 1.30±0.03, CCM3^+/-: 1.07±0.08) in control group mice brain, the relative expression of Claudin-5 (WT: 0.96±0.04, CCM3^+/-: 0.59±0.01) was decreased with statistical significance (F=199.27, P<0.001). The relative expression level of LRP-1 gene mRNA in brain of lead exposed group (WT: 0.32±0.10, CCM3^+/-: 0.06±0.01) was higher than that of unexposed group (WT:1.00±0.06, CCM3^+/-:2.12±0.18), the difference was statistically significant (F=288.29, P<0.001). The relative expression level of LRP-1 gene mRNA in brain of CCM3⁺/- mice exposed to lead was lower than that of WT mice ((0.06±0.01)vs(0.32±0.10), t=26.90, P<0.001). Conclusion The mice did not show significant AD-like lesions under low-does lead exposure, but resulted in early damage of brain blood-brain barrier and early changes of AD-like lesions in mice, with CCM3^+/- mice being sensitive to lead exposure stronger than that of WT mice, suggesting that deletion of CCM3 gene may be one of the potential risk factors for accelerating the development of AD in mice exposed to lead.

Objective To explore the impact of family education methods and parental rearing or not on the psychological problem tendency of primary school students.Methods The mental health test (MHT) was used on getting mental health data of the 2 838 primary school students from Dongguan city.Results (1) The results of MHT showed that 94.28％ of primary school students had no psychological problems,the tendency rate of psychological problems was 4.29％,and 1.43％ of them were suffering from serious psychological problems.Learning anxiety,physical symptoms and allergy tendency were more prominent.(2) The total MHT scores of primary school students with authoritarian,democratic,permissive and neglected family education methods were (34.84±14.58),(31.04±13.60),(35.19±12.82) and (41.19±13.10)respectively.There was no statistically significant difference in the terrorist tendency dimension scores of primary school students with different education methods (F=2.33,P=0.054),and the differences in the MHT total score and other dimensions were statistically significant (F=4.35-16.88,P＜0.01).(3) There were statistically significant differences in the scores of the total score of psychological problem tendency,dimensions of learning anxiety,anxiety to people,allergy tendency and impulse tendency of primary school students who were parental rearing or not (t=2.09-3.67,P＜0.05).However,there was no statistically significant difference in the dimensions of loneliness tendency,self-accusation tendency,physical symptoms and terrorist tendency (P＞0.05).Conclusion Family education methods and parenting styles has a impact on the mental health of primary school students.

OBJECTIVE: To investigate the effects of knockout cerebral cavernous malformation(CCM) virulence gene CCM3 on the migration induced by lead acetate in immortalized human umbilical vein endothelial cells(HUVECs) and to explore the possible mechanism of endoplasmic reticulum stress(ERS). METHODS: CCM3 wildtype(CCM3-WT) and CCM3 knockout(CCM3-KO) HUVECs were used as experimental cells. a) CCM3-WT and CCM3-KO HUVECs were treated with lead acetate at 0,10,50 and 200 μmol/L for 24 hours. The migration of these cells was observed by woundhealing assay. b) CCM3-WT and CCM3-KO HUVECs were treated with lead acetate at 0,10,50 and 200 μmol/L for 24 hours,and at 50 μmol/L for 0,6,12,24 and 48 hours,and the mRNA expression of genes of unfolded protein response pathway were detected by quantitative real-time polymerase chain reaction; the protein expression of glucose-regulated protein 78(GRP78) was detected by Western blotting. c) CCM3-WT and CCM3-KO HUVECs were divided into lead exposure group and tauroursodeoxycholic acid(TUDCA) group. The former was treated with 50 μmol/L lead acetate for 24 hours,and the latter was pre-treated with ERS inhibitor TUDCA,followed by 50 μmol/L lead acetate. The migration of these cells was observed by wound-healing assay. RESULTS: a) The migration of CCM3-WT and CCM3-KO cells decreased and showed a dose-effect relationship with the increase of lead acetate concentration(P < 0. 05). b) The mRNA relative expression of the GRP78,protein kinase-like endoplasmic reticulum kinase(PERK),transcription activator 4(ATF4) and CCAAT enhancer binding homologous protein(CHOP) in CCM3-KO cells treated with 10,50 and 200 μmol/L lead acetate were higher than that in CCM3-WT cells at the same doses,except for the GRP78 in CCM3-KO cells treated with10 μmol/L lead acetate(P < 0. 05). The mRNA expression of PERK and CHOP in CCM3-KO cells increased in a timeeffect relationship with the increase of lead-exposure time(P < 0. 05). The mRNA relative expression of the four genes in CCM3-KO cells were higher than those in CCM3-WT cells at 48 hours(P < 0. 05). When cells were treated with 50μmol/L lead acetate,the protein expression of GRP78 in CCM3-KO cells was higher than that in CCM3-WT cells(P <0. 05),and the protein expression of GRP78 in CCM3-KO cells increased in a time-effect relationship with the increase of lead-exposure time(P < 0. 05). c) The cell migration of TUDCA group was lower than that of lead-exposure group(P <0. 05). CONCLUSION: Lead acetate may activate ERS by activating the PERK-ATF4-CHOP signaling pathway,thereby reducing the migration of HUVECs. CCM3 gene has a protective effect on cell migration.

OBJECTIVETo investigate the effect of CCM3 gene defection on lead induced cell genotoxicity in mouse embryonic fibroblasts.

METHODSC57 female mice were mated with CCM3 gene heterozygous male mice. E13.5 embryos were taken to isolate primary mouse embryonic fibroblasts. After genotyping, wild type and heterozygous cells were treated with different doses of lead acetate. Cell viability, genotoxicity and protein expression were detected by MTS assay, CB micronucleus method and Western blot, respectively.

RESULTSMouse embryonic fibroblasts with lead acetate treatment for 24 h, wild-type cells 100.00 µmol/L lead acetate-treated group (69.16±1.36) and the control group (100.00±2.33) compared to cells decreased by 30%, CCM3 heterozygous type cell 100.00 µmol/L lead acetate-treated group (87.16±5.50) and the control group (100.00±2.06) compared to cells decreased by 13%, the difference was statistically significant (F values were 98.59, 82.63, P<0.001). Lead acetate treatment after 48 h, wild-type cells 100.00 µmol/L lead acetate-treated group (51.99±5.62) and the control group (100.00±3.11) compared to cells decreased by 50%, heterozygous type cells 100.00 µmol/L lead acetate treatment group (66.33±4.06) and the control group (100.00±5.72) compared to cells decreased by 35%, the differences were statistically significant (F values were 82.63, 36.86, P < 0.001). The results of CBMN test showed that with increased dose, micronucleus cell rate of two genotypes showed an increasing trend, in the wild-type cells, the micronucleus cell rate (/1 000) for the control group, 29.6±2.2, 6.25 µmol/L dose group 47.3±6.6, 25 µmol/L dose group 55.5±9.1, 100.00 µmol/L dose group 66.8±3.5; heterozygous cells micronucleus cell rate (/1 000) for the control group, 35.3±5.6, 6.25 µmol/L dose of 50.0±8.3, 25.00 µmol/L dose group 57.0±8.5, 100.00 µmol/L dose group 58.8±2.1. Micronucleus cell rates (/1 000) were significant differences, in 100.00 µmol/L dose groups of two genotypes. Western blot results showed that wild-type cells CCM3 expression 100.00 µmol/L lead acetate-treated group (0.70±0.03) was 1.32 times higher than the control group (0.53±0.07), heterozygous cells CCM3 expression 100.00 µmol/L lead acetate-treated group (0.48±0.02) was 1.77 times higher than control group that of 0.27±0.04, there was statistically significant difference (F values were 14.77, 25.74, P < 0.001); wild-type cells γ-H2AX expression 100.00 µmol/L lead acetate-treated group (0.69±0.03) was 1.06 times higher than the control group (0.65±0.07), heterozygous cells γ-H2AX expression 100.00 µmol/L lead acetate-treated group (0.99±0.04) was 1.55 times higher than the control group CCM3 expression levels (0.64±0.06), there was statistically significant difference (wild-type cells: F = 7.08, P = 0.012, heterozygous type cell: F = 13.49, P = 0.002).

CONCLUSIONCCM3 gene may play a role in lead-induced genetic toxicity of mouse embryonic fibroblasts, CCM3 gene-lead interactions effects on mouse embryonic fibroblasts cell toxicity.

Animals ; Apoptosis Regulatory Proteins ; DNA Damage ; Embryo, Mammalian ; Female ; Fibroblasts ; Genotype ; Male ; Membrane Proteins ; Mice ; Mice, Inbred Strains ; Micronuclei, Chromosome-Defective ; Organometallic Compounds ; Proto-Oncogene Proteins