BACKGROUND:Solute carrier family 1 member 5(SLC1A5)plays a potential role in a variety of diseases,but the exact mechanism of action is unclear.The construction of stable SLC1A5 overexpression and knockdown cell models can provide a powerful experimental tool for in-depth study of the exact role and mechanism of SLC1A5 in diseases and the discovery of potential therapeutic targets. OBJECTIVE:To construct lentiviral vectors for overexpression and knockdown of mouse SLC1A5 and establish stable transfected RAW264.7 cell lines,so as to provide an experimental foundation for further investigation of the role of SLC1A5 in inflammation. METHODS:Primers were designed and synthesized based on the SLC1A5 gene sequence,and the gene segment was amplified using polymerase chain reaction.Subsequently,the target gene segment was directionally inserted into the GV492 vector plasmid,which had been digested with AgeI/NheI enzymes,to construct recombinant lentiviral plasmids.Positive clones were further selected,and their sequences were confirmed.The pHelper1.0 plasmid vector and pHelper2.0 plasmid vector,along with the target plasmid vector,was co-cultured with 293T cells for transfection,resulting in the production and titration of lentiviral stocks.Furthermore,RAW264.7 cells were cultured in vitro,and the working concentration of puromycin was determined.Lentiviruses were separately co-cultured with RAW264.7 cells,and transfection efficiency was determined by measuring fluorescence intensity.Stable transfected cells were selected using puromycin,and real-time fluorescence quantitative PCR and western blot assay were employed to assess the gene and protein expression levels of SLC1A5 in stably transfected cell lines. RESULTS AND CONCLUSION:(1)Sequencing results indicated a perfect match between the sequencing and target sequences,confirming the successful construction of recombinant lentiviral vectors.(2)The titer for the overexpression SLC1A5 lentivirus was 1×109 TU/mL,while the titer for the knockdown SLC1A5 lentivirus was 3×109 TU/mL.(3)The working concentration of puromycin for RAW264.7 cells was determined to be 3 μg/mL.(4)The optimal conditions for transfecting RAW264.7 cells with overexpression/knockdown expression of SLC1A5 lentivirus involved the use of HiTransG P transfection enhancer with a multiplicity of infection value of 50.(5)A significant upregulation of the gene and protein expression levels of SLC1A5 was detected in cell lines stably overexpressing SLC1A5,while gene and protein expression levels of SLC1A5 were significantly decreased in the knockdown stable cell lines.These findings indicate that lentiviral vectors for mouse SLC1A5 overexpression and knockdown have been successfully constructed and a stably transfected RAW264.7 cell line has been obtained.

【Objective】 To explore the relationship between chronic comorbidity and the physical and mental health of relatives of elderly people during the nursing home confinement, and to analyze the mediating effects of perceived stress and intolerance of uncertainty in this context. 【Methods】 A total of 568 family members of elderly people in nine elderly institutions in Shaanxi Province were selected. The survey included the short version of the Perceived Stress Scale, Intolerance of Uncertainty Scale, and The World Health Organization-5 Well-being Index. The data were analyzed with Stata for correlation and mediation effects. 【Results】 ① The comorbidities of chronic diseases was positively correlated with the perceived stress (r=0.16, P<0.001) and intolerance of uncertainty (r=0.11, P=0.006) of the family members, but negatively correlated with the physical and mental health of the family members (r=-0.13, P=0.002). ② The mediating effect of perceived stress between chronic disease co-morbidity and physical and mental health of family members in older adults was -0.023, accounting for 18.8% of the total effect; the mediating effect of intolerance of uncertainty between chronic disease co-morbidity and physical and mental health of family members in older adults was -0.041, accounting for 33.5% of the total effect. 【Conclusion】 During closed management in a nursing facility, the physical and mental health of family members of older adults with chronic co-morbidities is poorer than that of family members of non-chronic co-morbidities. And it can lead to a decline in physical and mental health of family members through increased perceived stress and intolerance of uncertainty.

cFos is one of the most widely-studied genes in the field of neuroscience. Currently, there is no systematic database focusing on cFos in neuroscience. We developed a curated database—cFos-ANAB—a cFos-based web tool for exploring activated neurons and associated behaviors in rats and mice, comprising 398 brain nuclei and sub-nuclei, and five associated behaviors: pain, fear, feeding, aggression, and sexual behavior. Direct relationships among behaviors and nuclei (even cell types) under specific stimulating conditions were constructed based on cFos expression profiles extracted from original publications. Moreover, overlapping nuclei and sub-nuclei with potentially complex functions among different associated behaviors were emphasized, leading to results serving as important clues to the development of valid hypotheses for exploring as yet unknown circuits. Using the analysis function of cFos-ANAB, multi-layered pictures of networks and their relationships can quickly be explored depending on users’ purposes. These features provide a useful tool and good reference for early exploration in neuroscience. The cFos-ANAB database is available at www.cfos-db.net.

cFos is one of the most widely-studied genes in the field of neuroscience. Currently, there is no systematic database focusing on cFos in neuroscience. We developed a curated database-cFos-ANAB-a cFos-based web tool for exploring activated neurons and associated behaviors in rats and mice, comprising 398 brain nuclei and sub-nuclei, and five associated behaviors: pain, fear, feeding, aggression, and sexual behavior. Direct relationships among behaviors and nuclei (even cell types) under specific stimulating conditions were constructed based on cFos expression profiles extracted from original publications. Moreover, overlapping nuclei and sub-nuclei with potentially complex functions among different associated behaviors were emphasized, leading to results serving as important clues to the development of valid hypotheses for exploring as yet unknown circuits. Using the analysis function of cFos-ANAB, multi-layered pictures of networks and their relationships can quickly be explored depending on users' purposes. These features provide a useful tool and good reference for early exploration in neuroscience. The cFos-ANAB database is available at www.cfos-db.net .
Animals ; Fear ; Mice ; Neurons ; Proto-Oncogene Proteins c-fos ; Rats

Adaptation, Psychological ; Adolescent ; COVID-19 ; Child ; China ; Cross-Sectional Studies ; Female ; Humans ; Male ; SARS-CoV-2

Objective:To investigate the characteristics of consonant among children with speech sound disorder (SSD) and to provide an empirical basis for the subsequent clinical evaluation and evidence-based intervention.Methods:In this retrospective research a total of 1 395 children diagnosed with SSD from the Language-Speech Clinic of the Department of Children Health Care, Children′s Hospital, Capital Institute of Pediatrics from January 2007 to December 2018 were enrolled and underwent the phonological examination on the lexical level with picture naming, according to phoneme development in Chinese mandarin. The Chi-square trend test was applied to analyze the differences and trends of the proportion of consonant error subtypes in children of different age groups. The Chi-square test was conducted to compare the proportion of consonant error subtypes in different gender.Results:The 1 395 children diagnosed with SSD included 1 044 boys and 351 girls, with an age of (5.1±0.8) years. The occurrence of consonant errors in different locations of articulation was the highest for blade-alveolar /l/ (71.8%, 1 002/1 395) and the lowest for labial/b/(9.3%, 130/1 395). The occurrence of consonant errors of labial/p/f/, supra-dental/z/c/s/, blade-alveolar/t/l/, blade-palatal/ch/r/, velar/k/h/, and lingua-palatal/q/decreased with age (all P<0.05). The occurrence of consonant errors of labial/b/m/, supra-dental/z/c/, blade-alveolar/n/l/, blade-palatal/sh/, velar/h/, and lingua-palatal/x/were higher in boys than those in girls (10.3% (108/1 044) vs. 6.3% (22/351), 11.4% (119/1 044) vs. 6.0% (21/351), 64.8% (676/1 044) vs. 51.9% (182/351), 67.8% (708/1 044) vs. 59.8% (210/351), 16.7% (174/1 044) vs. 8.8% (31/351), 73.7% (769/1 044) vs. 66.1% (232/351), 58.0% (606/1 044) vs. 47.6% (167/351), 24.0% (251/1 044) vs. 14.2% (50/351), and 39.9% (417/1 044) vs. 27.6% (97/351); χ2 =5.167, 8.533, 16.889, 7.447, 12.863, 7.412, 11.650, 14.900, and 17.099, all P<0.05). The error types of consonant were substitution, omission, and distortion. Omission was the main error type of blade-alveolar/l/(53.3%, 743/1 395), distortion was the main error type of velar/h/(11.8%, 165/1 395), and substitution was the main error type of all other consonants. Substitution with blade-palatal/ch/having the highest occurrence of error (60.2%, 840/1 395). Conclusions:The occurrence of the blade-alveolar/l/error is the highest in children with SSD, with boys demonstrating more serious articulation problems. The main error type of consonant is substitution, with blade-palatal/ch/having the highest occurrence of error. These results suggest the necessity of attending to preschoolers′ articulation development. Phonological training targeting blade-alveolar/l/and blade-palatal/ch/should be carried out as early as possible.

Objective To explore influence of continued nursing on postoperative comprehensive ability of patients after craniotomy in neurosurgery department.Methods A total of 67 patients after craniotomy were randomly divided into experimental group with 35 cases and control group with 32 cases,among whom 3 lost the follow up and 34 patients and 30 were eventually included in the experimental group and the control group.The experimental group was given continued care and the control group applied conventional care after discharge.The comprehensive functional scores and self-assessment ability scores were recorded at 3 months after discharge.Results The experimental group had higher comprehensive functional scores at 3 months after discharge than the control group (P ＜ 0.05),physical function,and social cognitive score were higher in the experimental group (P ＜ 0.05),and no significant difference was found in verbal function between groups (P ＞ 0.05).Meanwhile,the self-care ability in the experimental group was higher than the control group (P ＜ 0.05).Conclusion The continued nursing model plays an important role in the recovery of patients after craniotomy.It can improve the patient's comprehensive function and self-care ability,and enhance quality of life of patients.

Objective To explore influence of continued nursing on postoperative comprehensive ability of patients after craniotomy in neurosurgery department.Methods A total of 67 patients after craniotomy were randomly divided into experimental group with 35 cases and control group with 32 cases,among whom 3 lost the follow up and 34 patients and 30 were eventually included in the experimental group and the control group.The experimental group was given continued care and the control group applied conventional care after discharge.The comprehensive functional scores and self-assessment ability scores were recorded at 3 months after discharge.Results The experimental group had higher comprehensive functional scores at 3 months after discharge than the control group (P ＜ 0.05),physical function,and social cognitive score were higher in the experimental group (P ＜ 0.05),and no significant difference was found in verbal function between groups (P ＞ 0.05).Meanwhile,the self-care ability in the experimental group was higher than the control group (P ＜ 0.05).Conclusion The continued nursing model plays an important role in the recovery of patients after craniotomy.It can improve the patient's comprehensive function and self-care ability,and enhance quality of life of patients.

Objective To explore the application value of circulating tumor cell (CTC) counting in the diagnosis of patients with colorectal cancer.Methods From July to October in 2015,a total of 61 patients firstly diagnosed with colorectal cancer,at stagc] to Ⅳ were enrolled.At the same period,20 individuals (healthy volunteers or patients with benign colorectal diseases) were selected as controls.Peripheral blood (7.5 mL) was taken before therapy.Peripheral blood CTC were counted by immunomagnetic beads enrichment combined with fluorescent staining method in two hours.The positive detection rate of CTC for colorectal cancer,especially early colorectal cancer was analyzed.And the priority of combination it with carcino-embryonic antigen (CEA) was also investigated.Chi-square test,t test and rank-sum test were used for statistical analysis.Results Two patients could not be pathological staged,due to the lack of whole body imaging evaluation.There were 30 patients with colorectal cancer at early stage (stage Ⅰ to Ⅱ) and 29 patients at advanced stage (stage Ⅲ to Ⅳ).The difference in CEA level between patients at early stage and advanced stage ((3.3±1.1) μg/L vs (20.4±3.3) μg/L) was statistically significant (t=-2.74,P=0.008).The median cell number of CTC of colorectal cancer group and control group was 1 (interquartile range=4) and 0,respectively,and the difference was statistically significant (Z=2.721,P=0.007).Taken 0/7.5 mL and 5 μg/L as the cut-off value of CTC and CEA,the detection rates of CTC for colorectal cancer and early colorectal cancer were 65.6％ (40/61) and 63.3％ (19/30),respectively;the detection rates of CEA for colorectal cancer and early colorectal cancer were 29.5 ％ (18/61) and 13.3％ (4/30).When CTC combined with CEA,the detection rates for colorectal cancer and early colorectal cancer were 73.8％ (45/61) and 66.7％ (20/30),respectively.Conclusions CTC has certain detection value in colorectal cancer especially early colorectal cancer.If it combined with CEA,the detection rate is much higher.

Objective To compare the clinical effects of two skin care regimens for incontinence-associated dermatitis. Methods A total of 53 patients with incontinence-associated dermatitis between March and September 2014 were randomly divided into two groups using random number table. Besides consistent skin care methods, group A was additionally treated with silicone coating dressing and group B was treated with skin protective liquid dressing at perineal skin area every 8 hours after cleaning. The effects of nursing care on healing rate and healing time of incontinence-associated dermatitis within 7 days of two groups were compared. Results The healing rate of group A was higher than that of group B (85. 71% vs 61. 54%). The difference between the two groups was statistically significant (χ2 =5. 352,P<0. 05). The healing time of group A was shorter than that of group B[(5.33 ±2.019)d vs (6.92 ±2.544)d]. The difference between the two groups was statistically significant (t= -2. 525,P<0. 05). The medical cost of group A was lower than that of group B[(480. 00 ± 181. 723) vs (623. 08 ± 228. 994)], the difference between the two groups was statistically significant (t= -2. 525, P<0. 05). Conclusions The effect of silicone coating dressing for the treatment of incontinence-associated dermatitis is superior to the skin protective liquid dressing.