Objective:To investigate the effects and molecular mechanism of non-coding RNA-activated DNA damage(NORAD)induced autophagy on oxaliplatin resistance in adenocarcinoma of esophagogastric junction (AEG).Methods:Four pairs of surgical samples of AEG and para-carcinoma normal tissues from patients with advance AEG treated in Anyang Tumor Hospital from January to June 2023 were collected. The expression of NORAD in AEG and para-carcinoma tissues was analyzed by long non-coding RNA microarray chip. The primary tumor cell line of AEG (PDC) was derived from fresh AEG tissues. Oxaliplatin-resistant cell lines of PDC and AEG cell line OE19 (PDC-R and OE19-R) were established. NORAD expression knockdown PDC-R and OE19 cell lines (shNORAD PDC-R and shNORAD OE19-R) were prepared by transfection. The target of NORAD, the correlation and interaction between microRNA-433-3p (miR-433-3p) and NORAD were predicted using Starbase v3.0 and DIANA-lncBase v3.0. PDC, PDC-R, OE19 and OE19-R cells were co-transfected with miR-144-3p and wild-type NORAD (NORAD-WT) or mutant NORAD (NORAD-Mut) plasmid, respectively. Dual-luciferase reporter assay was used to verify the correlation between NORAD and miR-433-3p. The expression levels of NORAD and miR-433-3p in normal gastric mucosal cell line GES-1 and AEG cell lines PDC, PDC-R, shNORAD PDC-R, OE19, OE19-R and shNORAD OE19-R were detected by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). The expression of p62 protein and microtubule-associated protein 1 light chain 3B-Ⅱ (LC3B-Ⅱ) was determined by Western blotting. The half inhibitory concentration (IC 50) of PDC, PDC-R, shNORAD PDC-R, OE19, OE19-R and shNORAD OE19-R cells was measured by cell counting kit-8 (CCK-8) assay. Independent sample t-test was used for statistical analysis. Results:The results of microarray analysis showed that NORAD was significantly up-regulated in AEG compared with that in para-carcinoma tissues (fold change≥2.0, P<0.05). Bioinformatics studies found that miR-433-3p was the potential target of NORAD. The results of dual-luciferase reporter assay indicated that the relative luciferase activity of the NORAD-WT group was lower than that of NORAD-Mut group in PDC and PDC-R cells (0.441±0.104 vs. 0.928±0.204, 0.449±0.112 vs. 0.947±0.201), and the differences were statistically significant ( t=-14.74 and -14.94, both P<0.001). The results of dual-luciferase reporter assay of OE19 and OE19-R cell lines were the same as those of PDC cell lines. The results of qRT-PCR showed that the expression of NORAD in GES-1 cells (1.016±0.213) was lower than that of PDC cells (2.194±0.322) and PDC-R cells (4.040±0.336), and the differences were statistically significant ( t=-14.94 and -37.21, both P<0.001). Furthermore, the expression level of NORAD in PDC was also found to be lower than that in PDC-R cells, and the difference was statistically significant ( t=-19.43, P<0.001). Additionally, shNORAD PDC-R cells exhibited lower expression level of NORAD (0.290±0.165) compared with PDC-R cells, and the difference was statistically significant ( t=-49.05, P<0.001). The expression level of miR-433-3p in GES-1 cells (1.017±0.248) was higher than that in PDC cells (0.470±0.156) and PDC-R cells (0.203±0.045), and the differences were statistically significant ( t=9.15 and 15.85, both P<0.001). Moreover, the expression level of miR-433-3p was found to be higher in PDC cells compared with PDC-R cells, and the difference was statistically significant ( t=8.11, P<0.001). Additionally, the expression level of miR-433-3p in shNORAD PDC-R cells (0.699±0.256) was also higher than that in PDC-R cells ( t=9.37, P<0.001). The results of Western blotting showed that the expression of LC3B-Ⅱ in PDC-R was higher than that in PDC cells (0.426±0.060 vs. 0.212±0.041), the expression of LC3B-Ⅱ in shNORAD PDC-R cells (0.155±0.029) was lower than that in PDC cells, and the differences were statistically significant ( t=8.70 and -79.45, both P<0.001). However the expression of p62 protein in each cell line showed an opposite trend, with a lower relative expression in PDC-R than PDC (0.205±0.031 vs. 0.311±0.400), and the expression in shNORAD PDC-R (0.504±0.084) was higher than that in PDC, and the differences were statistically significant ( t=-31.19 and 62.80, both P<0.001). The expression patterns of NORAD, miR-433-3p, LC3B-Ⅱ and p62 proteins in OE19, OE19-R and shNORAD OE19-R cells were similar to those in PDC. The results of CCK-8 assessment of target cell viability showed that the IC 50 values of PDC, PDC-R and shNORAD PDC-R cell lines were 14.28, 22.27 and 2.51 μg/mL, respectively; and the IC 50 values of OE19, OE19-R and shNORAD PDC-R cell lines were 3.95, 8.12 and 1.89 μg/mL, respectively. Conclusions:NORAD is highly expressed in AEG tissues and cells. NORAD is overexpressed in oxaliplatin-resistant cell lines and increase the autophagy activity of cells. After NORAD is knockdown, autophagy activity is inhibited and the sensitivity of AEG cells to oxaliplatin is significantly enhanced.

Objective:To determine the predictive value of serum neurofilament light chain (NfL) on neurologic function in out-of-hospital cardiac arrest (OHCA) patients.Methods:The clinical data of 96 OHCA patients admitted to Cangzhou Central Hospital from January 2018 to March 2022 were retrospectively analyzed. According to the Glasgow-Pittsburgh cerebral performance category (CPC) upon hospital discharge, the patients were divided into the favorable neurologic function (grade 1-2) and poor neurologic function (grade 3-5) groups. The difference of serum NfL was compared between the two groups, and the relationship between serum NfL and neurologic function was assessed using correlation analysis and logistic regression analysis. The area under the curve (AUC), sensitivity, and specificity of serum Nfl were calculated by receiver operating characteristic (ROC) curve. Hanley & McNeil method test was used to compare the difference of AUCs between serum NfL and neuron specific enolase (NSE).Results:Twenty-six percent (25/96) patients were discharged with favorable neurologic function. Serum NfL in the favorable neurological function group was significantly lower than that in the poor neurologic function group (47.6 pg/mL vs. 261.4 pg/mL, P<0.001). Correlation analysis showed that serum NfL was positively correlated with neurologic function ( r=0.69, P<0.001). Logistic regression analysis showed that serum NfL was independently associated with neurological function ( OR=0.92, 95% CI: 0.86-0.98; P=0.010). ROC curve indicated that the AUC of serum NfL in predicting poor neurologic function was 0.95 (95% CI: 0.92-0.99), with a sensitivity of 84.5% and a specificity of 100% at the cutoff value of 80.0 pg/mL. The AUC of serum NSE in predicting poor neurologic function was 0.79 (95% CI: 0.69-0.89), with a sensitivity of 67.6% and a specificity of 80.0% at the cutoff value of 45.1 ng/mL. A pairwise comparison using Hanley & McNeil method showed that the AUC of serum NfL in predicting poor neurologic function was higher than that of NSE ( Z=3.22, P=0.001). Conclusions:Serum NfL is helpful for clinician to predict neurologic function in OHCA patients.

ObjectiveTo observe the clinical efficacy and safety of traditional Chinese medicine （TCM） external therapeutic protocol of enriching pus for tissue growth （EPTG） in the treatment of Wagner 2-3 diabetic foot ulcer. MethodThe randomized controlled trial （RCT） design was adopted. Patients receiving basic treatment were divided into the EPTG group and the control group （debridement and change of nano-silver medical antibacterial dressing）. Ulcer healing rate， ulcer area， ulcer depth， TCM symptom score， visual analogue scale（VAS）， transcutaneous partial pressure of oxygen， wound blood flow， inflammatory factors ［C-reactive protein （CRP）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α）］， growth factors ［vascular endothelial growth factor （VEGF）， transforming growth factor-β₁ （TGF-β₁）， and fibroblast growth factor-β （FGF-β）］， adverse events， and outcome events of the two groups before and after treatment were observed. ResultCompared with the conditions before treatment， the ulcer healing rate， ulcer area， ulcer depth， TCM symptom score， VAS score， transcutaneous partial pressure of oxygen， wound blood flow， CRP， IL-6， TNF-α， VEGF， TGF-β₁， and FGF-β were significantly improved （P<0.01）. In terms of the improvement in the ulcer healing rate， ulcer area， ulcer depth， VAS score， transcutaneous partial pressure of oxygen， wound blood flow， CRP， IL-6， TNF-α， VEGF， TGF-β₁， and FGF-β， the EPTG group was superior to the control group （P<0.01）. There were no statistically significant differences in adverse events and outcome events between the two groups. ConclusionThe TCM external therapeutic protocol of EPTG is safe and effective in the treatment of Wagner 2-3 diabetic foot ulcer. It can greatly reduce the area and depth of diabetic foot ulcer， improve the ulcer healing rate and TCM symptom score， relieve the pain of patients， and improve the microcirculatory blood supply in the local ulcer. Its mechanism of action may be related to the reduction of the local inflammatory response of the wound and the improvement of the proliferation of fibroblasts and vascular endothelial cells.

Multiple plant lineages have independently evolved sex chromosomes and variable kary-otypes to maintain their sessile lifestyles through constant biological innovation.Morus notabilis,a dioecious mulberry species,has the fewest chromosomes among Morus spp.,but the genetic basis of sex determination and karyotype evolution in this species has not been identified.In this study,three high-quality genome assemblies were generated for Morus spp.[including dioecious M.notabilis(male and female)and Morus yunnanensis(female)]with genome sizes of 301-329 Mb and were grouped into six pseudochromosomes.Using a combination of genomic approaches,we found that the putative ancestral karyotype of Morus species was close to 14 protochromosomes,and that sev-eral chromosome fusion events resulted in descending dysploidy(2n=2x=12).We also charac-terized a～6.2-Mb sex-determining region on chromosome 3.Four potential male-specific genes,a partially duplicated DNA helicase gene(named MSDH)and three Ty3_Gypsy long terminal repeat retrotransposons(named MSTG1/2/3),were identified in the Y-linked area and considered to be strong candidate genes for sex determination or differentiation.Population genomic analysis showed that Guangdong accessions in China were genetically similar to Japanese accessions of mul-berry.In addition,genomic areas containing selective sweeps that distinguish domesticated mul-berry from wild populations in terms of flowering and disease resistance were identified.Our study provides an important genetic resource for sex identification research and molecular breeding in mulberry.

Objective:To investigate the correlation between radiation dose of the pelvic bone marrow of different anatomical bony sites and the incidence of neutrophil toxicity during the three-dimensional radiotherapy concurrent chemotherapy for cervical cancer.Methods:Clinical data of 117 cervical cancer patients who received three-dimensional radiotherapy concurrent chemotherapy from 2016 to 2018 were retrospectively analyzed. The dosimetric parameters included D mean, V 5Gy, V 10Gy, V 20Gy, V 30Gy, V 40Gy and V 50Gy. The correlation between the dosimetric parameters and the lowest neutrophil count was analyzed by linear regression analysis. Clinicopathological features and dosimetric parameters were included into the multivariate regression analysis model. Results:The incidence rates of neutrophil toxicity (grade 1-4) were 10.3%, 27.4%, 11.1% and 10.3%, respectively. The linear regression analysis showed that the D mean and V 50Gy of lumbosacral vertebrae (LS), the D mean, V 5Gy, V 10Gy, V 20Gy, V 30Gy, V 40Gy and V 50Gy of the ilium were significantly correlated with the grade 2-4 neutrophil toxicity ( P=0.035、<0.001、<0.001、=0.001、=0.003、=0.001、<0.001、<0.001、<0.001). Multivariate analysis demonstrated that the V 20Gy, V 30Gy and V 50Gy of the LS, the D mean, V 5Gy, V 10Gy, V 20Gy and V 30Gy of the ilium were significantly correlated with the grade 2-4 neutrophil toxicity ( P=0.046、0.038、0.049、0.041、0.039、0.029、0.036、0.029). Conclusion:During the process of three-dimensional radiotherapy concurrent chemotherapy for cervical cancer, the volume of medium-and high-dose of LS and the volume of low-and medium-dose of ilium are significantly correlated with the risk of neutrophil toxicity.

Objective:To study the orthopedic treatment strategy for hemophilia complicated with musculoskeletal disorders as well as the peri-operative consumption of clotting factor.Methods:Total 338 orthopedic surgeries were performed for 261 patients, average age of 30.6 y (6-65 y) , with hemophilia between January 1996 and December 2019 at our institute. Two hundred and twenty-six patients presented with bleeds within the joints. Sixty-one patients presented with intramuscular bleeds, 45 presented with hemophilic pseudotumors, and six presented with miscellaneous complaints. Strategy of clotting factor replacement therapy was designed as per differences in the level of the operation procedure. Information regarding clinical manifestation, operative strategy, clotting factor consumption, and re-operation for complications was retrospectively recorded. The costs for multiple joint procedure and single joint procedure were studied.Results:We found that 270 of the 338 surgical procedures were major surgical procedures (79.9%) . There were 203 procedures of joint arthroplasty (60%) . Fourteen patients underwent reoperations for local recurrence (4.2%) . The average factor Ⅷ consumption before the surgery was 44.4 ± 8.1 IU/kg. The average FⅧ consumption within postoperative 2 weeks was 40 962 IU (647±177 IU/kg) . Seven type A hemophilic patients developed F Ⅷ inhibitor following the surgical procedure, with an average level of 13.7±11.2 BU/mL. Sixty-eight patients underwent multiple joint procedures under one anesthesia session (26%) . There was no significant difference in the factor consumption between the multiple joint procedure and single joint procedure.Conclusions:Surgical treatment was found to be effective for hemophilic arthropathy and lesion of the musculoskeletal apparatus, with the clotting factor replacement therapy. Multiple joint procedures under one anesthesia were more cost effective for patients with hemophilia, with less factor consumption than staged single joint procedure.

Objective To investigate the effect of long non-coding RNA HOX transcript antisense RNA ( lncRNA HOTAIR ) on the cell proliferation, radiosensitivity and apoptosis of the rectal adenocarcinoma cell lines SW480 and HCT116 in vitro. Methods The expression levels of lncRNA HOTAIR in the rectal adenocarcinoma cell lines ( SW480 and HCT116 ) were assessed by real-time quantitative PCR (RT-qPCR).Silencing of HOTAIR by RNA interference was performed to explore its roles in cell proliferation,radiosensitivity and apoptosis. After treatment with irradiation at a gradient dose,the cell viability was measured and the rate of cell apoptosis was tested. Results Compared with the human rectal epithelial cell lines,the expression of lncRNA HOTAIR was significantly higher in the rectal adenocarcinoma cell lines. The colonic assay demonstrated that the sensitizing enhancement ratios ( SERs) were 1. 58 and 1. 33 for the cells transfected with HOTAIR siRNA in SW480 and HCT116 cell line compared with the control isRNA transfection group. In vitro silencing of lncRNA HOTAIR could enhance the apotosis rate and radiosensitivity of the rectal adenocarcinoma cell line SW480. Conclusion The expression level of lncRNA HOTAIR is correlated with the cellular radiosensitivity, which is probably a parameter for predicting the radiosensitivity of rectal adenocarcinoma cells. Radiotherapy combined with HOTAIR-siRNA can significantly inhibit the cell proliferation,induce cell apoptosis and enhance the radiosensitivity.

Objective To evaluate the changes of CEA,CA19-9,CA72-4 of patients with advanced adenocarcinoma of the esophagogastric junction (AEGJ) who received ultra-early precise hyperthermic intraperitoneal perfusion chemotherapy (HIPEC) with paclitaxel (PTX) after radical surgery.Methods From Jul 2014 to Dec 2015 postoperative advanced AEGJ patients were divided to receive ultraearly precise HIPEC (87 cases,HIPEC group) and conventional therapy group (85 cases) in early stage after radical surgery.Serum levels of CEA,CA19-9 and CA72-4 were examined on preoperative day 1 and postoperative day 9.Results On post-op day 9,the level of CEA,CA19-9 and CA72-4 was (2.8 ± 1.3)ng/ml,(22 ±8)IU/ml,(4.1 ± 1.9)IU/ml in HIPEC group,while that was (3.4 ± 1.2)ng/ml,(25 ±11) IU/ml,(4.8 ± 2.1) IU/ml in control group respectively (t =2.453,P =0.015;t =2.241,P =0.026;t =2.154,P =0.033).Conclusion Ultra-early PTX-based HIPEC with advanced AEGJ after radical surgery lowers serum tumor makers of CEA,CA19-9,CA72-4,reducing body's tumor burden.

Spinal cord injury is a catastrophic injury causing lifelong severe disabilities, and poses a great burden to the individuals, families and society. In order to promote the standardization in treatment of traumatic spinal cord injury, the consensus on the evaluation, treatment and rehabilitation of traumatic spinal cord injury was suggested by experts, who came from authoritative multicenter in China. The expert consensus, which formed a standardization process from the first aid clinical treatment to rehabilitation of spinal cord injury, shall give a better practical guide for clinic and rehabilitation physicians.