Objective:To reveal the molecular characteristics of mononuclear phagocytes (MPs) in rat model of peripheral nerve injury (PNI) using single-cell RNA sequencing (scRNA-seq) technology that would provide the developmental changes and major biological process involved in the function of MPs after PNI.Methods:Twenty-seven male SD rats (200-300 g in weight) were selected from the Department of Hand and Foot Microscopy and Wound Repair Surgery, Henan Provincial People's Hospital (People's Hospital of Zhengzhou University) and the Department of Orthopaedics of First Affiliated Hospital of Zhengzhou University from July 2023 to December 2023. The rats were divided into a Sham operation group (Sham group), a 3 days post crush group (3 dpc group) and a 7 days post crush group (7 dpc group), following the randomised table method with 9 rats per group. After 7 days of environmental acclimatisation, the 3 dpc group and 7 dpc group were subjected to have the right sciatic nerve crushed in order to create a model of crush injury. And as a control group, the Sham group was subjected to Sham surgery only. Nine right sciatic nerves of rats were collected from each group at the corresponding time pints. Single-cell isolation was performed on the 10X Genomics platform. ScRNA-seq libraries were constructed using the Gel Bead Kit V3 and the libraries were sequenced using an Illumina Novaseq 6000 sequencer. Dimensionality reduction was performed using Principal Component Analysis and T-Distributed Stochastic Neighbor Embedding to visualise and explore the cellular heterogeneity within the dataset. Nine distinct cell clusters and their corresponding marker genes were identified based on the dimensionality-reduced data. Differential gene expression analysis was then performed to identify differentially expressed genes (DEGs) in MPs between different groups. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed to uncover the biological processes and pathways based on the DEGs. Monocle program for pseudo-time analysis was used to infer the developmental trajectory of MPs after injury.Results:A total of 19 054 cells were obtained by sequencing, and the results showed that the proportion of MPs in peripheral nerves was significantly up-regulated after PNI, and MPs were classified into 9 cellular subgroups based on the clustering analysis of the scRNA-seq data, which were Cluster 1 (3 398 cells), Cluster 2 (3 388 cells), Cluster 3 (3 262 cells), Cluster 4 (2 825 cells), Cluster 5 (2 753 cells), Cluster 6 (1 894 cells), Cluster 7 (648 cells), Cluster 8 (492 cells) and Cluster 9 (394 cells), respectively. Based on the expression of different cell subpopulation markers, MPs in the Sham group, 3 dpc group and 7 dpc group of sciatic nerves were classified into 9 cell clusters and the distributions of different MPs clusters in the 9 sciatic nerve samples were identified, among which, the Sham group had the lowest number of MPs cells in the sciatic nerve samples (a total of 2 719 cells) and the clusters were mainly dominated by clusters 5 (1 119 cells) and clusters 6 (1 240 cells). The 3 dpc group had the highest number of MPs cells (9 760 cells in total) and the clusters were mainly dominated by cluster 2 (1 760 cells), cluster 3 (3 130 cells) and cluster 4 (2 300 cells). The MPs (6 575 cells in total) in the 7 dpc group were mainly dominated by cluster 1 (2 406 cells) and cluster 2 (1 628 cells). Compared with the Sham group, the GO and KEGG annotations of the DEGs were significantly upregulated in the 3 dpc group, indicating that MPs in the rat sciatic nerves would have the ability to bind to extracellular molecules and remove debris from the injury site at 3 days post-injury, and the 7 dpc group would have the ability to activate the signalling pathways related to nerve repair. The proposed time-series analysis revealed that, in the uninjured condition, the MPs were mainly in the cluster 5 (Ccl17 +Cd80 +) and cluster 6 (Fcmr +Slc9a9 +). At 3 days post-injury, MPs developed into cell types dominated by cluster 2 (Cd8b +Meis3 +), cluster 3 (Il10 +Cd163 +) and cluster 4 (Ccl24 +Prg4 +). At 7 days post-injury, the effector state of cluster 2 among the main cell types of MPs was still maintained but the other parts had developed into cluster 1 (Hspa1b +Apobec1 +) related phenotypes. Conclusion:The molecular characteristics of MPs in the peripheral nerve revealed through scRNA-seq data provide valuable insights into the role of MPs in mediating inflammation and neural regeneration after PNI.

Objective:To explore the independent risk factor for in-hospital mortality of patients with multiple trauma, and to construct a prediction model of risk of death and validate its efficacy.Methods:A retrospective cohort study was performed to analyze the clinical data of 1 028 patients with multiple trauma admitted to Affiliated Hospital of Jiangsu University from January 2011 to December 2021. There were 765 males and 263 females, aged 18-91 years[(53.8±12.4)years]. The injury severity score (ISS) was 16-57 points [(26.3±7.6)points]. There were 153 deaths and 875 survivals. A total of 777 patients were enrolled as the training set from January 2011 to December 2018 for building the prediction model, while another 251 patients were enrolled as validation set from January 2019 to December 2021. According to the outcomes, the training set was divided into the non-survival group (115 patients) and survival group (662 patients). The two groups were compared in terms of the gender, age, underlying disease, injury mechanism, head and neck injury, maxillofacial injury, chest injury, abdominal injury, extremity and pelvis injury, body surface injury, damage control surgery, pre-hospital time, number of injury sites, Glasgow coma score (GCS), ISS, shock index, and laboratory test results within 6 hours on admission, including blood lactate acid, white blood cell counts, neutrophil to lymphocyte ratio (NLR), platelet counts, hemoglobin, activated partial thromboplastin time (APTT), fibrinogen, D-dimer and blood glucose. Univariate analysis and multivariate Logistic regression analysis were performed to determine the independent risk factors for in-hospital mortality in patients with multiple trauma. The R software was used to establish a nomogram prediction model based on the above risk factors. Area under the receiver operating characteristic (ROC) curve (AUC), calibration curve and clinical decision curve analysis (DCA) were plotted in the training set and the validation set, and Hosmer-Lemeshow goodness-of-fit test was performed.Results:Univariate analysis showed that abdominal injury, extremity and pelvis injury, damage control surgery, GCS, ISS, shock index, blood lactic acid, white blood cell counts, NLR, platelet counts, hemoglobin, APTT, fibrinogen, D-dimer and blood glucose were correlated with in-hospital mortality in patients with multiple trauma ( P<0.05 or 0.01). Logistic regression analysis showed that GCS≤8 points ( OR=1.99, 95% CI 1.12,3.53), ISS>25 points ( OR=7.39, 95% CI 3.50, 15.61), shock index>1.0 ( OR=3.43, 95% CI 1.94,6.08), blood lactic acid>2 mmol/L ( OR=9.84, 95% CI 4.97, 19.51), fibrinogen≤1.5 g/L ( OR=2.57, 95% CI 1.39,4.74) and blood glucose>10 mmol/L ( OR=3.49, 95% CI 2.03, 5.99) were significantly correlated with their in-hospital mortality ( P<0.05 or 0.01). The ROC of the nomogram prediction model indicated that AUC of the training set was 0.91 (95% CI 0.87, 0.93) and AUC of the validation set was 0.90 (95% CI 0.84, 0.95). The calibration curve showed that the predicted probability was consistent with the actual situation in both the training set and validation set. DCA showed that the nomogram prediction model presented excellent performance in predicting in-hospital mortality. In Hosmer-Lemeshow goodness-of-fit test, χ2 value of the training set was 9.69 ( P>0.05), with validation set of 9.16 ( P>0.05). Conclusions:GCS≤8 points, ISS>25 points, shock index>1.0, blood lactic acid>2 mmol/L, fibrinogen≤1.5 g/L and blood glucose>10 mmol/L are independent risk factors for in-hospital mortality in patients with multiple trauma. The nomogram prediction model based on these 6 predictive variables shows a good predictive performance, which can help clinicians comprehensively assess the patient′s condition and identify the high-risk population.

Strain-resource engineering is often considered as an important infrastructure of microbiology related research and industry. The western developed countries took the lead in establishing the classical microbial resource utilization method, and continuously improved the preservation system, species annotation technology and global sharing mechanism, which realized the expansion and reserve of biological resources since end of the 19th century. The rich and diversified germplasm resources, standard strains and production strains not only have important economic values, but also maintain the advantages of scientific research, bioeconomy (such as antimicrobial agents, vaccines, detection reagent development and standard development, etc.) and national security. Although there has been a lot of progress in related research in recent years, compared with developed countries, there is still a big gap in related fields in China. The investment and top-level design in this area lag far behind the western developed countries, and it is not commensurate with the current level of economic and social development in my country. Drawing lessons from the practice of WFCC and WDCM (World Data Center for Microorganisms, Global microbial data Center, affiliated to WFCC), for the purpose of collecting new clinical species/strains, this paper puts forward some suggestions on the identification, preservation and upload system of isolates.

Objective:To analyze the effects of DHT on the proliferation and migration of endothelial progenitor cells (EPCs) and the role of RhoA/ROCK pathway in this process.Methods:Early EPCs were isolated from peripheral blood of healthy adults, and cultured in serum-free EBM-2 medium for 24 h before incubation with various concentrations of DHT (1, 10, and 100 nmol/L). EPCs proliferative and migrative capacities were measured. The adherent cells were collected and randomLy divided into: control group, DHT group, C3 exoenzyme+DHT, Y-27632+DHT group. EPCs proliferation and migration were assayed by MTT assay and modified Boyden chamber assay respectively.Results:DHT significantly increased the proliferation and migration ability of EPCs in a dose- and time-dependent manner, maximum at 10 nmol/L, 24 h ( P<0.05). C3 exoenzyme [(0.22±0.02) vs (0.26±0.05), P>0.05] and Y-27632 [(0.21±0.04) vs (0.26±0.05), P>0.05] can attenuate the proliferative capacities of EPCs induced by DHT compared with the DHT group, but there was no statistical significance. The influence of DHT on EPCs migrative capacities can be abolished by C3 exoenzyme [(35.26±4.27) vs (46.92±5.46), P<0.05] and Y-27632 [(33.61±5.33) vs (46.92±5.46), P<0.01]. C3 exoenzyme [(116.75±7.42) vs (156.80± 21.74), P<0.05] and Y-27632 [(121.73±5.33) vs (156.80 ±21.74), P<0.01] could noticeably attenuate DHT-induced EPCs secretion of VEGF respectively. Conclusions:DHT can modulate EPCs proliferation, migration and the RhoA/ROCK pathway plays an important role in this process.

Objective:To evaluate the protective efficacy and safety of Brucella 104M against aerosol challenge in BALB/c mice and characterize its immunological effects. Methods:Female mice of 6-8 weeks old were immunized with Brucella abortus strain 104M by intratracheal aerosol delivery or intranasal instillation or subcutaneous injection route. Six mice of each group were sacrificed at 4, 8, 16, 24 weeks after immunization. At each time point, the clinical manifestations of mice were investigated, the serum, spleen and lung samples of mice were collected, body weight, spleen weight, bacteria loads in spleens, the anti- Brucella antibodies titers in serum and the cytokines concentrations of IFN-γ, IL-18 in serum or lung homogenate of the mice were detected. Twenty two weeks after immunization, all the mice were challenged with Brucella A19 through intratracheal aerosol delivery. Results:Compared with the control group, neither abnormal clinical symptoms nor significant changes in body weight were found in 104M immunization groups, at each time point when immunized through either nose dropping route, subcutaneous injection or aerosol routes; and the spleen weight of immunization groups were lower than control group after challenge ( P<0.05): *M1 (0.26±0.16)g

Objective:To compare the wound healing time, Surgical site infection (SSI) rate and other postoperative outcomes between the gunsight closure and purse-string closure technique in loop stoma closure.Methods:Between November 2013 and December 2017, a total of 143 patients who underwent gunsight stoma reversal were included in this multicenter prospective randomized controlled trial. The patients were randomized to undergo gunsight (gunsight group, n=72) or purse-string closure technique (purse-string group, n=71). The primary endpoint was wound healing time. The second endpoints were the incidence of SSI, morbidity, and patient satisfaction. Statistical analysis between groups was performed using the t-test, repeated measures analysis of variance, Mann-Whitney U test, χ 2 test or Fisher′s exact test. Results:There were 45 males and 27 females with age of 67 (11) (M( Q R)) years in gunsight group, 42 males and 29 females with age of 65 (20) years in purse-string group. The body mass index, American Society of Anesthesiologist classification, comorbidities, primary diagnosis, the type of ostomy, intraoperative blood loss, perioperative complications, postoperative hospital stay, hospitalization cost, SSI rate and incisional hernia (stoma site) between the 2 groups were not significantly different ( P>0.05). Although had a statistically longer operating time (80(10) minutes vs. 70(10) minutes, Z=-2.381, P=0.017), patients who underwent gunsight procedure and a significantly shorter wound healing time (17(2) days vs. 25(4) days, Z=-10.199, P<0.01), higher patient satisfaction score with regards to wound healing time (3(1) vs. 3(1), Z=-4.526, P<0.01), and higher total patient satisfaction score (25(3) vs. 25(3), Z=-2.529, P=0.011) compared with those who underwent purse-string procedure. Conclusions:The gunsight and purse-string techniques are effective procedures for stoma reversal and both have low SSI rate. The gunsight technique is associated with shorter wound healing time, higher levels of patient satisfaction compared with purse-string technique, and is recommended as the closure technique of choice.

Objective:To compare the wound healing time, Surgical site infection (SSI) rate and other postoperative outcomes between the gunsight closure and purse-string closure technique in loop stoma closure.Methods:Between November 2013 and December 2017, a total of 143 patients who underwent gunsight stoma reversal were included in this multicenter prospective randomized controlled trial. The patients were randomized to undergo gunsight (gunsight group, n=72) or purse-string closure technique (purse-string group, n=71). The primary endpoint was wound healing time. The second endpoints were the incidence of SSI, morbidity, and patient satisfaction. Statistical analysis between groups was performed using the t-test, repeated measures analysis of variance, Mann-Whitney U test, χ 2 test or Fisher′s exact test. Results:There were 45 males and 27 females with age of 67 (11) (M( Q R)) years in gunsight group, 42 males and 29 females with age of 65 (20) years in purse-string group. The body mass index, American Society of Anesthesiologist classification, comorbidities, primary diagnosis, the type of ostomy, intraoperative blood loss, perioperative complications, postoperative hospital stay, hospitalization cost, SSI rate and incisional hernia (stoma site) between the 2 groups were not significantly different ( P>0.05). Although had a statistically longer operating time (80(10) minutes vs. 70(10) minutes, Z=-2.381, P=0.017), patients who underwent gunsight procedure and a significantly shorter wound healing time (17(2) days vs. 25(4) days, Z=-10.199, P<0.01), higher patient satisfaction score with regards to wound healing time (3(1) vs. 3(1), Z=-4.526, P<0.01), and higher total patient satisfaction score (25(3) vs. 25(3), Z=-2.529, P=0.011) compared with those who underwent purse-string procedure. Conclusions:The gunsight and purse-string techniques are effective procedures for stoma reversal and both have low SSI rate. The gunsight technique is associated with shorter wound healing time, higher levels of patient satisfaction compared with purse-string technique, and is recommended as the closure technique of choice.

Objective To investigate the risk factors of polytrauma combined with multiple organ dysfunction syndrome (MODS).Methods A retrospective case control study was performed on the clinical data of 299 polytrauma patients admitted to the Affiliated Hospital of Jiangsu University from December 2011 to June 2017.The collected information included gender,age,length of hospital stay,number of injured parts,injury severity scores (ISS),neutrophil count,leukocyte level,hemoglobin level,platelet count,activated partial thromboplastin time (APTI),and D-dimer level within 24 hours since admission.In addition,shock within 24 hours since admission,infection after 3 days since admission,damage control surgery,underlying diseases and prognostic outcomes were also recorded.All the patients were divided into MODS group (94 patients) and non-MODS group (205 patients).Univariate and multivariate logistic regression analyses were used to determine the risk factors of polytrauma combined with MODS.Receiver operating characteristic (ROC) curve was applied to further analyze those risk factors identified by the former analyses.Results In the univariate analysis,there were statistically significant differences between the two groups in the number of injured parts,ISS,hemoglobin level,platelet count,APTT,D-dimer level within 24 hours since admission,shock within 24 hours since admission,infection after 3 days since admission,damage control surgery and prognostic outcomes (P ＜ 0.05).No significant differences were found in gender,age,underlying disease,length of hospital stay,neutrophil level,the leukocyte level within 24 hours since admission between the two groups (P ＞ 0.05).The multivariate logistic regression analysis showed that ISS (OR =1.048),shock within 24 hours since admission (OR =3.913),infection after 3 days since admission (OR =27.715),and D-dimer level within 24 hours since admission (OR =1.015) were significantly associated with polytrauma combined with MODS (P ＜ 0.05).In addition,the area under ROC curve of ISS was 0.726 (95 ％ CI 0.667-0.784),and the area under ROC curve of D-dimer was 0.638 (95％ CI 0.571-0.706).Conclusions The risk factors of polytrauma patients combined with MODS include ISS,infection after 3 days since admission,D-dimer level and shock within 24 hours since admission.In the treatment of polytrauma patients,attention should be paid to assessment of injury severity and coagulation function,active resuscitation to correct shock,prevent and control infection,which can reduce and prevent the risks for polytrauma patients combined with MODS.

OBJECTIVETo apply single nucleotide polymorphism microarray (SNP array) for the detection of genome-wide copy number variations(CNVs) in fetuses with malformations and women with an adverse reproductive history, and to explore the correlation of rare CNVs with the clinical manifestations.

METHODSAmniotic fluid and umbilical cord blood samples were collected from 314 women with singleton pregnancy. SNP array was performed on samples where chromosomal abnormalities were excluded after G-banding analysis.

RESULTSPathological CNVs were detected in 8.91% (28/314) of all samples, which included 11 duplications, 9 deletions, 4 loss of heterozygosity (LOH), and 4 conjoined deletions and duplications. The sizes of duplications and deletions were between 0.47 Mb and 16.7 Mb, and between 0.16 Mb and 13.3 Mb, respectively. Fifteen CNVs were mapped to the regions of microdeletion or microduplication syndromes or regions associated with clinical manifestations, while the remainder 13 were considered benign or variant of uncertain significance.

CONCLUSIONA proportion of fetuses with malformations and women with an adverse reproductive history may be attributed to CNVs, half of which are mapped with to the regions of well known syndromes. SNP array may facilitate discovery of new syndromes and provide a basis for genetic counseling and prenatal diagnosis.

Adult ; Chromosome Aberrations ; Chromosome Disorders ; diagnosis ; embryology ; genetics ; DNA Copy Number Variations ; Female ; Fetal Diseases ; diagnosis ; genetics ; Humans ; Infant ; Infant, Newborn ; Male ; Polymorphism, Single Nucleotide ; Pregnancy ; Pregnancy Complications ; diagnosis ; genetics ; Prenatal Diagnosis ; Reproductive History ; Young Adult

Objective: To explore the plasma chemokines expressions and related clinical implication in patients with Stanford type A aortic dissection (AD). Methods: We retrospectively analyzed the data of 65 patients with Stanford type A aortic dissection, hypertensive patients and 11 healthy subjects admitted in our department from October 2013 to December 2014, they were divided into four groups: NH-CON group (11 healthy subjects), H-AD group (29 AD patients with hypertension), NH-AD group (21 AD patients without hypertension), and H-CON group (14 hypertension patients). Four plasma samples from AD patients and 4 plasma samples from healthy subjects were collected randomly with random numbers table, and the levels of different chemokines were examined by protein array analysis. Then, plasma levels of chemokines including macrophage inflammatory protein 1β(MIP-1β), epithelial neutrophil activating peptide 78(ENA-78), interleukin 16(IL-16), interferon inducible protein 10(IP-10) and FMS-like tyrosine kinase 3(Flt-3) ligand were analyzed by luminex. Pearson analysis was used to determine the correlations between the chemokines and serum C reactive protein (CRP) levels. Results: Plasma levels of MIP-1β(34.0(29.3, 47.2) ng/L vs. 51.0(28.2, 80.7) ng/L, P<0.05) and ENA-78(110.5(59.1, 161.4) ng/L vs. 475.7(299.3, 837.3) ng/L, P<0.05) were significantly lower in H-AD group, while plasma IL-16 level was significantly higher in H-AD group(54.7(16.3, 187.8) ng/L vs. 17.5(11.9, 20.8) ng/L, P<0.05) than in H-CON group. Plasma levels of MIP-1β(48.3(26.4, 62.1) ng/L, P<0.05) were significantly lower in H-AD patients than in NH-AD patients. Plasma level of ENA-78 was significantly lower in NH-AD group than in NH-CON group (95.0(58.0, 155.0) ng/L vs. 257.7(85.2, 397.8) ng/L, P<0.05). The levels of IP-10 and Flt-3 ligand were similar among the 4 groups (all P>0.05). Pearson analysis showed that there were no correlation between MIP-1β(r²=0.01, P>0.05), ENA-78(r²=0.02, P>0.05), IL-16(r²=0.02, P>0.05), IP-10(r²=0.00, P>0.05), Flt-3 ligand(r²=0.02, P>0.05) and CRP levels in patients with Stanford type A aortic dissection. Conclusions Lower plasma levels of MIP-1β and ENA-78 and higher plasma levels of IL-16 may associate with the occurrence and development of type A aortic dissection, but their concentrations are not correlated with serum CRP levels. There is no significant change on plasma levels of IP-10 and Flt-3 in the Stanford type A aortic dissection patients.