Forensic examination materials are often plagued by trace amounts,mixes,and other factors.Single-cell isolation technology can solve these forensic problems to some extent by studying each cell individually to obtain comprehensive and reliable information.There are many single cell isolation techniques available in research reports,such as flow cytometry,laser capture microdissection,etc.This review will summarize the most common single cell isolation techniques used by researchers today,and summarize the application of various techniques in forensic science,summarize the selection strategies for single-cell isolation techniques in different scenarios based on cost,degree of automation,yield,cell damage rate,and the availability of relevant forensic platforms,and finally explore the forensic application prospects of single-cell isolation techniques.In general,single cell isolation can be applied to multiple fields such as mixed stain examination,post-mortem time inference,pre-and post-mortem injury determination,forensic toxicology analysis,forensic microbiology and forensic anthropology.The development of single cell isolation technology is of great value to the application of forensic medicine,and will provide a new way of deciphering difficult examination materials.

Objective:To explore the standardized performance of a FISH probe before clinical detection.Methods:The probe sensitivity and specificity of ETV6/RUNX1 were analyzed via interphase and metaphase FISH in 20 discarded healthy bone marrow samples. The threshold system of the probe was established using an inverse beta distribution, and an interpretation standard was established. Finally, a parallel-controlled polymerase chain reaction detection study was conducted on 286 bone marrow samples from patients at our hospital. The clinical sensitivity, specificity, and diagnostic coincidence rate of ETV6/RUNX1 FISH detection were analyzed, and the diagnostic consistency of the two methods was analyzed by the kappa test.Results:The probe sensitivity and specificity of the ETV6/RUNX1 probe were 98.47% and 100%, respectively. When 50, 100, and 200 cells were counted, the typical positive signal pattern cutoffs were 5.81%, 2.95%, and 1.49%, respectively, and the atypical positive signal pattern cutoffs were 13.98%, 9.75%, and 6.26%, respectively. The clinical sensitivity of FISH was 96.1%, clinical specificity was 99.6%, diagnostic coincidence rate was 99.00%, diagnostic consistency test kappa value was 0.964, and P value was <0.001.Conclusion:For FISH probes without a national medical device registration certificate, standardized performance verification and methodology performance verification can be performed using laboratory developed test verification standards to ensure a reliable and accurate reference basis for clinical diagnosis and treatment.

Objective:To analyze the quality control test results of urine fluoride laboratories in Jiangxi Province, and support reliable data for the monitoring and prevention of fluorosis.Methods:From 2019 to 2021, the Jiangxi provincial urine fluoride laboratories used the "Determination of Fluoride in Urine-Ion Selective Electrode Method" (WS/T 89-2015) to detect quality control samples. Origin 9 software was used to analyze the data, and Youden plots and deviation bar plots were produced. The results were evaluated based on the Z-score (qualified: │Z│ ≤2, basically qualified: 2 < │Z│ < 3, unqualified: │Z│ ≥3).Results:The qualified rate of quality control assessment results of urine fluoride laboratories in Jiangxi Province was 95.00% (38/40) . Most of urine fluoride laboratories had scores of │Z│ < 2, and two laboratories had scores │Z│≥3 in 2019 and 2020.Conclusion:The quality control assessment results of urine fluoride laboratories in Jiangxi Province are relatively satisfactory, but some laboratories still need to further improve their detection capabilities.

Hereditary gingival fibromatosis (HGF) is a rare inherited condition with fibromatoid hyperplasia of the gingival tissue that exhibits great genetic heterogeneity. Five distinct loci related to non-syndromic HGF have been identified; however, only two disease-causing genes, SOS1 and REST, inducing HGF have been identified at two loci, GINGF1 and GINGF5, respectively. Here, based on a family pedigree with 26 members, including nine patients with HGF, we identified double heterozygous pathogenic mutations in the ZNF513 (c.C748T, p.R250W) and KIF3C (c.G1229A, p.R410H) genes within the GINGF3 locus related to HGF. Functional studies demonstrated that the ZNF513 p.R250W and KIF3C p.R410H variants significantly increased the expression of ZNF513 and KIF3C in vitro and in vivo. ZNF513, a transcription factor, binds to KIF3C exon 1 and participates in the positive regulation of KIF3C expression in gingival fibroblasts. Furthermore, a knock-in mouse model confirmed that heterozygous or homozygous mutations within Zfp513 (p.R250W) or Kif3c (p.R412H) alone do not led to clear phenotypes with gingival fibromatosis, whereas the double mutations led to gingival hyperplasia phenotypes. In addition, we found that ZNF513 binds to the SOS1 promoter and plays an important positive role in regulating the expression of SOS1. Moreover, the KIF3C p.R410H mutation could activate the PI3K and KCNQ1 potassium channels. ZNF513 combined with KIF3C regulates gingival fibroblast proliferation, migration, and fibrosis response via the PI3K/AKT/mTOR and Ras/Raf/MEK/ERK pathways. In summary, these results demonstrate ZNF513 + KIF3C as an important genetic combination in HGF manifestation and suggest that ZNF513 mutation may be a major risk factor for HGF.
Animals ; Humans ; Mice ; Fibromatosis, Gingival/pathology* ; Gingiva ; Kinesins/genetics* ; Mutation/genetics* ; Phosphatidylinositol 3-Kinases/genetics*

Hip fractures are among the most common fractures in the elderly, presenting to be a leading cause of disability and mortality. Surgical treatment is currently the main treatment method for hip fractures. The incidence of perioperative malnutrition is increased after hip fractures in the elderly due to the comorbidities, decreased basal metabolic rate, accelerated protein breakdown, weakened anabolism and surgical stress. However, malnutrition not only increases the incidence of postoperative complications, but also leads to increased mortality, indicating an important role of perioperative nursing management of nutrition for the elderly patients with hip fractures. At present, there still lacks scientific guidance and application standards on perioperative nursing management of nutrition for the elderly patients with hip fractures. Therefore, the Orthopedic Nursing Committee of Chinese Nursing Association and the Editorial Board of Chinese Journal of Trauma organized relevant experts to formulate the Expert consensus on perioperative nursing management of nutrition for elderly patients with hip fractures ( version 2023) according to evidence-based medical evidences and their clinical experiences. Fourteen recommendations were made from aspects of nutrition screening, nutrition assessment, nutrition diagnosis, nutrition intervention and nutrition monitoring to provide guidance for perioperative nursing management of nutrition in elderly patients with hip fractures.

Objective:To analyze the etiological characteristics and molecular epidemiological correlation of five cases of typhoid fever during the same period in yantai city.Methods:Six S. Typhis strains were isolated from 5 typhoid patients and epidemiological samples in Yantai city in 2018. The onset time of the cases were from May 26, 2018 to July 24, 2018, distributed in Shuidao Town of Muping District, Dengzhou Street of Penglai District, Donglai Street of Longkou District, Wenhua Street of Muping District and Fulaishan Street of zhifu District. S. Typhis strains were analyzed by conventional bacterial isolation method and XbaⅠ/BlnⅠ double-enzyme digestion pulse-field gel electrophoresis (PFGE). Meanwhile, ViaB virulence gene detection and 27 common antibiotics sensitivity tests were conducted to study the etiology of S. Typhis.Results:Six strains of S. Typhi were isolated from 5 patients and the domestic egg of one patient, which were divided into 4 PFGE patterns by PFGE-XbaⅠ and PFGE-BlnⅠ and among which 3 strains had the same PFGE patterns.One multi-drug resistant strain (foreign patient), one single-drug resistant strain (patient with a history of provincial retention), and one completely sensitive strain were detected. The three strains of the same PFGE pattern exhibit the same drug-sensitive phenotype which were intermediate against aminoglycosides and quinolones and susceptibility against the other antibiotics.All of the strains carried the ViaB virulence factor except the strain from the foreign patient.Conclusion:Local S. Typhi is susceptibility or intermediate against antibiotics commonly used in clinic.Sporadic cases of typhoid fever and typhoid imported infections still need attention.

Objective: To compare the clinical outcomes of fresh embryo transfers (ETs) and frozen-thawed embryo transfers (FETs) after a freeze-all cycle in women ≥40 years old with poor ovarian response (POR). Methods: We performed a single-center, retrospective, case-control study of patients who underwent in vitro fertilization between January 2014 and June 2019. We included a total of 192 patients aged 40 years or older from whom 3 or fewer oocytes had been retrieved and who were receiving cleavage-stage ET in this study. Of these patients, 101 and 91 patients underwent fresh ET and FET, respectively. The primary outcome was the live birth rate (LBR) after the first ET. Logistic regression analysis was used to compare the IVF outcomes and neonatal characteristics between the fresh ET and FET groups, adjusting for maternal age, body mass index, luteinizing hormone, and the number of good quality embryos transferred. Results: The mean maternal ages and number of oocytes retrieved (43.2 years and 2.3 in both groups, P=0.902 and P=0.927, respectively) were similar in the fresh ET and FET groups. No significant difference was observed between the LBRs of the fresh ET and FET groups (adjusted odds ratio, 1.28; 95% confidence interval, 0.29–5.70). The clinical pregnancy and miscarriage rates, and neonatal characteristics (birth weights and premature infant rates) were similar between the 2 groups. Conclusions FET after the freeze-all strategy had no beneficial impact on the clinical outcomes of women ≥40 years with POR.

Objective: To compare the clinical outcomes of fresh embryo transfers (ETs) and frozen-thawed embryo transfers (FETs) after a freeze-all cycle in women ≥40 years old with poor ovarian response (POR). Methods: We performed a single-center, retrospective, case-control study of patients who underwent in vitro fertilization between January 2014 and June 2019. We included a total of 192 patients aged 40 years or older from whom 3 or fewer oocytes had been retrieved and who were receiving cleavage-stage ET in this study. Of these patients, 101 and 91 patients underwent fresh ET and FET, respectively. The primary outcome was the live birth rate (LBR) after the first ET. Logistic regression analysis was used to compare the IVF outcomes and neonatal characteristics between the fresh ET and FET groups, adjusting for maternal age, body mass index, luteinizing hormone, and the number of good quality embryos transferred. Results: The mean maternal ages and number of oocytes retrieved (43.2 years and 2.3 in both groups, P=0.902 and P=0.927, respectively) were similar in the fresh ET and FET groups. No significant difference was observed between the LBRs of the fresh ET and FET groups (adjusted odds ratio, 1.28; 95% confidence interval, 0.29–5.70). The clinical pregnancy and miscarriage rates, and neonatal characteristics (birth weights and premature infant rates) were similar between the 2 groups. Conclusions FET after the freeze-all strategy had no beneficial impact on the clinical outcomes of women ≥40 years with POR.

Objective:To analyze the etiological characteristics and molecular epidemiological correlation of five cases of typhoid fever during the same period in yantai city.Methods:Six S. Typhis strains were isolated from 5 typhoid patients and epidemiological samples in Yantai city in 2018. The onset time of the cases were from May 26, 2018 to July 24, 2018, distributed in Shuidao Town of Muping District, Dengzhou Street of Penglai District, Donglai Street of Longkou District, Wenhua Street of Muping District and Fulaishan Street of zhifu District. S. Typhis strains were analyzed by conventional bacterial isolation method and XbaⅠ/BlnⅠ double-enzyme digestion pulse-field gel electrophoresis (PFGE). Meanwhile, ViaB virulence gene detection and 27 common antibiotics sensitivity tests were conducted to study the etiology of S. Typhis.Results:Six strains of S. Typhi were isolated from 5 patients and the domestic egg of one patient, which were divided into 4 PFGE patterns by PFGE-XbaⅠ and PFGE-BlnⅠ and among which 3 strains had the same PFGE patterns.One multi-drug resistant strain (foreign patient), one single-drug resistant strain (patient with a history of provincial retention), and one completely sensitive strain were detected. The three strains of the same PFGE pattern exhibit the same drug-sensitive phenotype which were intermediate against aminoglycosides and quinolones and susceptibility against the other antibiotics.All of the strains carried the ViaB virulence factor except the strain from the foreign patient.Conclusion:Local S. Typhi is susceptibility or intermediate against antibiotics commonly used in clinic.Sporadic cases of typhoid fever and typhoid imported infections still need attention.

Objective　A group of patients with instability of walking as the main manifestation and combined with hypofolateemia were collected and their clinical characteristics were summarized. Methods　54 patients with folic acid <3.1 IU/ml，without cerebrovascular disease，whose first symptom manifested as "walking instability" were classified as the first group with folic acid deficiency in walking instability. Folic acid <3.1 IU/ml combined with cerebrovascular disease was listed as the control group of folic acid reduction combined with cerebrovascular disease，a total of 129 patients. The characteristics of baseline data，biochemical metabolism data，and brain MRI brain ischemic lesion distribution were compared between the two groups，and Logistic multivariate regression analysis was performed. Results　The onset of folic acid deficiency in the onset of walking instability was （7.56±8.78） days，which was longer than the folic acid deficiency combined cerebrovascular disease control group at （5.19±6.87） days，t=4.586，P=0.001.The onset age of folic acid deficiency in the first manifestation of walking instability was （77.11±9.97） years，which was higher than the folic acid deficiency combined cerebrovascular disease control group at （73.30±12.42） years，t=2.906，P=0.022.The homocysteine in the folic acid deficiency group at the first manifestation of unstable walking was （19.88±9.00），which was lower than the folic acid deficiency combined with cerebrovascular disease control group （30.38±59.77），t=2.938，P=0.021.Focal deficiencies in the folic acid-deficient group at the onset of walking instability were higher （41.67%） than in the control group （33.72%），χ2=6.254，P=0.012.Instability of walking was first manifested in the folic acid-deficient group，and most of the ischemic foci in the cerebral hemisphere were spot-shaped and independent. Folate deficiency combined with cerebrovascular disease control group，most of the cerebral hemisphere ischemic lesions were fused into linear，band-shaped lesions with blurred borders. Using the folic acid deficiency group as the dependent variable for the first manifestation of walking instability，a binary logistic multivariate regression analysis revealed that the cerebral lobe ischemic focus was OR=2.311，95%CI 1.174～4.569，P=0.025.Conclusion　Patients with instability in walking and folic acid deficiency may have a longer onset，older age，and insignificant increase in homocysteine levels. A cranial MRI examination showed frontal，temporal and parietal lobes. The number of cerebral lobe ischemic foci is relatively large，and they are distributed in a dot-like and independent manner.