Objective:To construct the sphingosine kinase 1(SPHK1)overexpression lentiviral vector,and to establish the SKOV3 lentiviral stable transfection cell line.Methods:According to the SPHK1 data information provided by the National Center for Biotechnology Information(NCBI)database,the primers were designed and synthesized,the target gene was amplified,and connected to the GV492 plasmid treated with Bam HⅠ and AgeⅠ restriction enzymes to construct the SPHK1 overexpression lentiviral vector;the positive clones were selected for PCR and sequencing identification;the lentiviral plasmid and the lentiviral packaging auxiliary plasmid were co-transfected into the HEK-293T cells for packaging and titer determination;according to the measured optimal multiplicity of infection(MOI)of 10,the corresponding lentiviral amounts in various groups were transfected into the SKOV3 cells,and the SKOV3 cells were divided into blank group(without treatment),GV492 control group(GV492 control lentivirus infected SKOV3 cells),and GV492-SPHK1 overexpression group(GV492-SPHK1 overexpression lentivirus infected SKOV3 cells,ov-SPHK1 group);the optimal concentration of 2 mg·L-1 puromycin was used to screen the stably transfected SKOV3 cell line;after 48 h,the medium was changed and replaced with 1 mg·L-1 puromycin for screening for 14 d;the morphology and fluorescence expression of the cells were observed under fluorescence microscope;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of SPHK1 mRNA in the SKOV3 cells in various groups;Western blotting method was used to detect the expression level of SPHK1 protein in the SKOV3 cells in various groups.Results:The PCR sequencing results showed that the gene sequence of the SPHK1 overexpression lentiviral vector was completely consistent with the target sequence,and the SPHK1 overexpression lentiviral vector was successfully constructed;the titer determination results showed that the lentiviral titers in GV492 control group and ov-SPHK1 group were 5×1011 and 8×1011 TU·L?1,respectively;the SKOV3 cells in GV492 control group and ov-SPHK1 group were in good state and showed strong fluorescence expression,suggesting that the SKOV3 stable transfection cell line overexpressing SPHK1 was successfully established;the RT-qPCR results showed that compared with blank group and GV492 control group,the expression level of SPHK1 mRNA in the SKOV3 cells in ov-SPHK1 group was significantly increased(P<0.01);the Western blotting results showed that compared with blank group and GV492 control group,the expression level of SPHK1 protein in the SKOV3 cells in ov-SPHK1 group was significantly increased(P<0.01).Conclusion:The SPHK1 overexpression lentiviral vector is successfully constructed,and the SKOV3 stable transfection cell line is established.

Parvalbumin interneurons belong to the major types of GABAergic interneurons. Although the distribution and pathological alterations of parvalbumin interneuron somata have been widely studied, the distribution and vulnerability of the neurites and fibers extending from parvalbumin interneurons have not been detailly interrogated. Through the Cre recombinase-reporter system, we visualized parvalbumin-positive fibers and thoroughly investigated their spatial distribution in the mouse brain. We found that parvalbumin fibers are widely distributed in the brain with specific morphological characteristics in different regions, among which the cortex and thalamus exhibited the most intense parvalbumin signals. In regions such as the striatum and optic tract, even long-range thick parvalbumin projections were detected. Furthermore, in mouse models of temporal lobe epilepsy and Parkinson's disease, parvalbumin fibers suffered both massive and subtle morphological alterations. Our study provides an overview of parvalbumin fibers in the brain and emphasizes the potential pathological implications of parvalbumin fiber alterations.
Mice ; Animals ; Epilepsy, Temporal Lobe/pathology* ; Parvalbumins/metabolism* ; Parkinson Disease/pathology* ; Neurons/metabolism* ; Interneurons/physiology* ; Disease Models, Animal ; Brain/pathology*

Objective: To analyze the situation of new occupational diseases in Hengyang City from 2006 to 2017, and put forward prevention and control strategies. Methods: The data of the new occupational disease report of Hengyang city in the Occupational Disease and Occupational Health Information Monitoring System from January 1, 2006 to December 31, 2017 was collected, and the age, working years, and the region and industry of the new occupational disease patients were analyzed. Results: From 2006 to 2017, there were 7 categories, 30 kinds and 2 110 cases of new occupational diseases in Hengyang City, including 1 117 cases of pneumoconiosis, 951 cases of chronic occupational poisoning, 15 cases of acute occupational poisoning, 12 cases of occupational otolaryngological and stomatological diseases, 7 cases of occupational skin diseases, 6 cases of occupational diseases caused by physical factors, and 1 case of occupational eye diseases(cataracts), 1 case of occupational tumor (lung cancer and skin cancer caused by arsenic and its compounds). New occupational diseases were mainly concentrated in Changning and Leiyang County-level city (87.82%, 1 853/2110), among which occupational poisoning had the most incidence in Changning County-level city (97.83%, 945/966), and pneumoconiosis had the most incidence in Leiyang County-level city (67.05%, 749/1 117). New occupational diseases were mainly concentrated in the manufacturing and mining industries (95.59%, 2 017/2 110). Pneumoconiosis (63.74%, 712/1 117) and acute occupational poisoning (60.00%, 9/15) were mainly caused by small businesses. Chronic occupational poisoning (61.62%, 586/951) and occupational otolaryngological and stomatological diseases (75.00%, 9/12) were mainly caused by large enterprises. Conclusion The new occupational diseases in Hengyang city are mostly pneumoconiosis and chronic occupational poisoning, and we should focus on strengthening the prevention and control of occupational diseases in the mining industry and manufacturing industry.

Objective To explore the correlation between polymorphisms (SNPs) of locus rs2269272 of glial high-affinity glutamate transporter (SLC1A3) gene and suicide attempt among Chinese adolescents.Methods iMLDRTM genotyping technology was used to detect the polymorphism of rs2269272 SLC1A3 gene loci in 55 suicide attempters and 112 healthy controls,and survival analysis was applied to analyze the relationship between allele (T) and the age of suicide attempt.Results The differences of rs2269272 locus allele distribution between two groups were statistically significant(x2=4.208,P=0.040),but genotype distribution of two groups had no significantly differences(x2 =4.011,P=0.135).Non-suicidal self-injury adolescents with locus rs2269272 (15.6 ± 0.4) were younger than adolescents without locus rs2269272(16.4±0.7),but the difference was not statistically significant.Conclusion Preliminary findings suggest that rs2269272 SLC1 A3 may be relevant to non-suicidal self-injury acts,and rs2269272 locus allele is not related to earlier suicide attempt.