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 compare the effect of anesthesia mode on the neurological functional outcomes in patients undergoing endovascular treatment for acute posterior circulation ischemic stroke.Methods:Clinical data of 656 patients undergoing intravascular therapy for acute posterior circulation ischemic stroke registered in online Acute Stroke Patients for Stroke Management Quality Evaluation Database from January 2017 to December 2022 were retrospectively analyzed.The data included 163 cases with conscious sedation and 493 cases with general anesthesia during the procedure.After propensity score matching,428 patients were included in the analysis,including 155 cases in the conscious sedation group and 273 cases in the general anesthesia group.The differences of operation mode,etiology type,vascular recanalization,hemorrhagic transformation at 24 h,modified Rankin Scale(mRS)score at 3 months and mortality within 3 months were compared between the two groups.Binary logistic regression was used to explore the effect of different anesthesia mode on neurological functional outcomes.Results:There was a significant difference in operation mode between the two groups(P<0.01),while there were no significant differences in etiology type,vascular recanalization,hemorrhagic transformation at 24 h,mRS score at 3 months or mortality within 3 months(all P>0.05).Binary logistic regression analysis revealed that anesthesia modes were not significantly associated with functional outcomes of patients(OR=1.151,95%CI:0.751-1.765,P>0.05).Conclusion:Anesthesia mode(conscious sedation or general anesthesia)will not affect the neurological functional outcomes in patients with acute posterior circulation ischemic stroke undergoing endovascular treatment.

Objective:To construct a thirst management scheme for adult surgical patients and provide practical guidance for nurses to carry out perioperative thirst management.Methods:From April to May 2023, literature search was conducted to extract domestic and foreign evidence related to thirst, and after group discussion, a preliminary management plan for thirst in adult surgical patients was constructed. From May to June 2023, 18 experts were selected for two rounds of Delphi expert letter consultation, and a formal plan for thirst management of adult surgical patients was discussed and revised according to expert opinions.Results:A total of 18 experts completed two rounds of expert letter consultation. The effective recovery rate of the questionnaire was 100.00% and the expert authority coefficient was 0.915. The coefficients of variation for the importance and operability scores of the first round of expert inquiry were 0.05-0.25 and 0.06-0.42, respectively, and the Kendall's harmony coefficients were 0.224 and 0.184 ( P<0.01). The coefficients of variation for the importance and operability scores of the second round of inquiry were 0.05-0.23 and 0.06-0.24, respectively, and the Kendall's harmony coefficients were 0.166 and 0.154 ( P<0.01). The final thirst management scheme included five primary items (preliminary preparation, identification and evaluation, preoperative relief strategies, postoperative intervention measures and effectiveness evaluation) and 23 secondary items. Conclusions:The thirst management scheme for adult surgical patients constructed in this study is scientific and comprehensive, which can provide a reference basis for clinical practice.

Objective:To explore the value of amide proton transfer-weighted (APTw) imaging and intravoxel incoherent motion (IVIM) imaging for diagnosing and evaluating the pathological differentiation of cervix squamous cell carcinoma (CSCC).Methods:This study was a diagnostic trial. Totally 56 patients pathologically diagnosed with CSCC at the First Hospital of Lanzhou University from October 2021 to October 2022 were retrospectively collected, as the CSCC group. And 36 female healthy volunteers who underwent physical examinations at the First Hospital of Lanzhou University from October 2021 to October 2023 were recruited as the control group. CSCC patients were divided into well-moderately differentiated ( n=34) and poorly differentiated groups ( n=22). The region of interest was placed in the lesions of CSCC group and normal cervical stroma of control group, and the quantitative parameters for asymmetric magnetization transfer ratio (MTR asym) of APTw imaging and pure diffusion coefficient (D), false diffusion coefficient (D *) and perfusion fraction (f) for IVIM were obtained. The independent sample t test was used to compare the differences in quantitative parameters between the two groups, the logistic regression model was used to establish combined parameters for the quantitative parameters with statistical significance between the two groups. The receiver operator characteristic curve was used to evaluate the diagnostic efficacy of single quantitative parameters and combined parameters to distinguish the CSCC group from the control group, and the well-moderately differentiated group from the poorly differentiated group in CSCC patients. The area under the curve (AUC) was compared using the DeLong test. Results:There were significant differences in MTR asym, D and f between CSCC group and control group ( t=-9.79, 10.09, 11.35, P<0.001). Also, significant differences were found for MTR asym and D between the well-moderately differentiated and poorly differentiated group ( t=4.11, -3.76, P<0.001). There was no significant difference in other quantitative parameters ( P>0.05). When comparing the CSCC group and control group, the AUC (95% CI) of MTR asym, D, f and combined parameter (MTR asym+D+f) were 0.887 (0.804-0.944), 0.940 (0.871-0.979), 0.968 (0.909-0.993), 0.995 (0.950-1.000). The AUC of the combined parameter was higher than those of MTR asym and D, with statistical significance ( Z=3.07, 2.06, P=0.002, 0.040). When comparing the well-moderately differentiated and poorly differentiated group, the AUC (95% CI) of MTR asym, D, and combined parameter (MTR asym+D) were 0.789 (0.660-0.887), 0.775 (0.644-0.876), 0.852 (0.731-0.932). There was no significant difference between each two AUCs ( P>0.05). Conclusion:The quantitative parameters of APTw and IVIM imaging can be used to diagnose and preliminarily evaluate the pathological differentiation of CSCC. Joint parameters can improve the diagnostic efficiency of CSCC.

Paperless management of medical records is of great significance in promoting refined hospital management. Driven by national policies and the development of information technology in medical institutions, paperless medical records have become an inevitable trend in the development of medical records. However, at present, there were still a large number of medical record data in China that cannot be digitized or directly uploaded to hospital electronic medical record systems. Since 2019, the First Hospital of Jiaxing has launched a paperless medical record construction project and established a paperless medical record management system, forming a closed-loop management system for the entire process of medical record collection, archiving, storage, management, and utilization. By the end of 2021, paperless of medical records in all departments had been achieved, electronic signatures had been fully implemented, and paperless versions of reports of standalone equipment, high-value consumables barcodes, surgical anesthesia related documents, and outsourced inspection reports had been achieved, with a paperless rate of 98.5%. Through paperless management of medical records, the supervision of medical record quality and medical quality has been strengthened, operating costs have been reduced and work efficiency has been improved. At the same time, this has provided convenience for medical personnel to access medical record information and applicate of the data, and has effectively improved the hospital′s level of refined management. It can provide reference for paperless management of medical records in other medical institutions.

Objective:To explore the clinical characteristics and prognostic influencing factors of multiple primary lung cancer (MPLC), and to construct a prognostic prediction model.Methods:The clinical data and prognostic information of MPLC patients diagnosed by pathological examination included in the Surveillance, Epidemiology, and End Results (SEER) database from January 2010 to December 2020 were retrospectively analyzed. Patients were randomly divided into training and validation sets according to a 7:3 ratio using R software. Survival curves were plotted by using the Kaplan-Meier method and log-rank test was used for comparison between groups. The independent influencing factors of overall survival (OS) of MPLC patients in the training set were screened using univariate and multivariate Cox proportional hazards models, and accordingly, the nomogram predicting the survival rate of patients at 3, 5 and 8 years were plotted. In the training and validation sets, using the actual survival as the gold standard, the receiver operating characteristic (ROC) curves of the constructed models for predicting the patients' 3-, 5- and 8-year OS rates were plotted, the area under the curve (AUC) was obtained, and C-index of the model was analyzed by using R software. The calibration curves of 3-, 5- and 8-year OS rates predicted by the models and the actual OS rates were plotted.Results:A total of 5 495 MPLC patients were included, 3 846 in the training set and 1 649 in the validation set. The differences in the composition of patients of different ages and AJCC stages between the training and validation sets were statistically significant (both P < 0.05), and the differences in the comparison of other clinicopathological characteristics were not statistically significant (all P > 0.05). The results of multivariate Cox regression analysis showed that males (compared with females, HR = 1.256, 95% CI: 1.144-1.379, P < 0.001), age ≥ 70 years old (compared with 50-59 years old, HR = 1.201, 95% CI: 1.030-1.400, P = 0.019), FPLC with pathological types of squamous cell carcinoma or other types (compared with adenocarcinoma, HR = 1.275, 95% CI: 1.137-1.431, P < 0.001; HR = 1.208, 95% CI: 1.041-1.403, P = 0.013), and SPLC with pathological types of squamous cell carcinoma, small cell lung carcinoma, or other types (compared with adenocarcinoma, HR = 1.270, 95% CI: 1.121-1.440, P < 0.001; HR = 1.978, 95% CI: 1.642-2.384, P < 0.001; HR = 1.246, 95% CI: 1.090-1.424, P = 0.001), and AJCC stage Ⅲ and Ⅳ (compared with stage Ⅰ, HR = 1.645, 95% CI: 1.447-1.869, P < 0.001; HR = 2.078, 95% CI: 1.669-2.587, P < 0.001), FPLC without operation (compared with operation, HR = 1.263, 95% CI: 1.038-1.536, P = 0.020), SPLC without operation (operation vs. no operation, HR = 0.680, 95% CI: 0.579-0.799, P < 0.001), FPLC without lymph node dissection or with clearance of 1-3 regional lymph nodes (compared with clearance of ≥4, HR = 1.225, 95% CI: 1.016-1.477, P = 0.034; HR = 1.314, 95% CI: 1.103-1.566, P = 0.002), FPLC with maximum diameter 3-5 cm or >5 cm (compared with <3 cm, HR = 1.181, 95% CI: 1.053-1.324, P = 0.005; HR = 1.232, 95% CI: 1.069-1.420, P = 0.004), and SPLC with maximum diameter 3-5 cm or >5 cm (compared with <3 cm, HR = 1.560, 95% CI: 1.362-1.786, P < 0.001; HR = 1.727, 95% CI: 1.451-2.054, P < 0.001), and FPLC without chemotherapy (chemotherapy vs. no chemotherapy or unknown, HR = 0.744, 95% CI: 0.655-0.845, P < 0.001) were the independent risk factors of patients' poor OS (all P < 0.05). The results of Kaplan-Meier survival analysis showed that the OS of patients with different gender, race, age, two tumor locations, AJCC staging, pathological type of two lung tumors, maximum diameter of two tumors, and whether two tumors were treated surgically or not, and whether two tumors were treated with chemotherapy or not in the training set were compared, and the differences were all statistically significant (all P < 0.05). Based on the independent factors affecting the OS of MPLC patients screened by the results of multivariate Cox regression analysis, nomogram predicting the 3-, 5- and 8-year OS rates of MPLC were plotted. The results of ROC curve analysis showed that the C-index of the training set's nomogram was 0.679 (95% CI: 0.649-0.701), and the AUC values for predicting the 3-, 5- and 8-year OS rates were 0.601, 0.595 and 0.586, respectively; the C-index of the validation set was 0.678 (95% CI: 0.633-0.720), and the AUC values for predicting 3-, 5- and 8-year OS rates were 0.643, 0.631 and 0.626, respectively. The calibration curves showed that the 3-, 5- and 8-year OS rates of patients predicted by the nomogram models in both the training and validation sets were in good agreement with the actual results with a high goodness-of-fit. Conclusions:The established prognostic model has good predictive value and can effectively assess the prognosis of patients.

Objective:To investigate the change in 25-hydroxy vitamin D (25-OHD) levels in hospitalized newborns in the neonatal intensive care unit (NICU) between baseline and vitamin D supplementation, and to explore the effect of different levels of vitamin D on the complications.Method:A prospective study was conducted on the newborns admitted to NICU at Jingzhou Hospital Affiliated to Yangtze University within 72 h after birth from January 2021 to January 2022. Vitamin D supplementation was initiated after the detection of basal 25-OHD levels within 72 h after birth. Serum 25-OHD levels were measured after 2, 4, and 6 weeks of supplementation. Newborns were categorized into four groups according to the basal 25-OHD level: sufficient, insufficient, deficient, and severely deficient groups. The analysis of variants, independent sample t-test, paired sample t-test, Chi-square test, or Fisher's exact probability method were employed to evaluate the differences in basal 25-OHD levels among newborns with different clinical conditions and gestational ages, as well as the variation in 25-OHD levels before and after supplementation among the four groups. Furthermore, differences in the morbidity and mortality among different basal status groups were analyzed. Result:(1) During the study period, 626 cases met the inclusion criteria, and after excluding seven cases, 619 infants were ultimately included in the study with serum 25-OHD level within 72 h being (21.8±10.1) ng/ml. There were 134 cases (21.6%) in the sufficient group, 208 cases (33.6%) in the insufficient group, 186 cases (30.0%) in the deficient group, and 91 cases (14.7%) in the severe deficient group. (2) No statistically significant differences were observed in the basal 25-OHD levels regardless of the genders, gestational age, birth month, number of fetuses or small for gestational age (all P>0.05). (3) Among all infants, 158 cases continued to supplement vitamin D for two weeks, 64 cases continued for four weeks, and 13 cases continued for six weeks, with all of them discharged within eight weeks. Compared with the basal 25-OHD levels, there were no statistically significant differences in the serum 25-OHD levels among the sufficient, insufficient, deficient, and severely deficient groups after two weeks of supplementation [(37.1±9.3) vs. (36.8±4.9) ng/ml, (24.7±7.2) vs. (24.7±2.9) ng/ml, (16.0±7.6) vs. (15.4±2.9) ng/ml, (8.1±5.6) vs. (7.6±1.4) ng/ml; t=0.18, 0.04, 0.65 and 0.48, respectively; all P>0.05]. After four weeks of supplementation, however, the serum 25-OHD levels in the four groups were higher than those before supplementation [(40.0±5.2) vs. (35.8±3.9) ng/ml, (29.7±6.4) vs. (24.5±2.9) ng/ml, (20.3±7.1) vs. (15.6±3.0) ng/ml, (14.9±7.3) vs. (6.5±2.3) ng/ml; t=2.13, 2.66, 5.08 and 7.64, respectively; all P<0.05]. (4) The incidence of hypocalcemia [23.1% (21/91) vs. 9.7% (18/186)] and neonatal respiratory distress syndrome [15.4% (14/91) vs. 3.2% (6/186)] were higher in the severely deficient group than those in the deficient group ( χ2=9.07 and 13.49, both P<0.008). No statistically significant differences were observed in the incidence of neonatal sepsis, neonatal necrotizing enterocolitis, bronchopulmonary dysplasia, retinopathy of prematurity, and mortality among the four groups (all P>0.05). Conclusions:The insufficiency of 25-OHD levels and vitamin D deficiency were prevalent in NICU neonates. Vitamin D status did not significantly differ among newborns with varying gestational ages. A prolonged period of sustained vitamin D supplementation may be required to elevate the serum 25-OHD level. The incidence of hypocalcemia and neonatal respiratory distress syndrome are higher in newborns with severe vitamin D deficiency.

RNA ; CRISPR-Cas Systems

【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.