Objective To study the impact of ethanol in nucleic acid extraction products on quantitative real-time PCR(qPCR), and to provide ethanol concentration limits for the extraction quality of more sensitive nucleic acid detection methods.Methods A potassium dichromate oxidation method was developed to detect the ethanol residues in the products extracted by two commercialized magnetic bead nucleic acid extraction kits, testing whether the ethanol residues impact qPCR. Then different concentrations of ethanol solutions were mixed with qPCR reagents that had been dissolved in simulated extraction products and freeze-dried(hereinafter referred to as freeze-dried reagents) for amplification. The sensitivity changes were observed, and the Ct values, fluorescence gain intensity and amplification efficiency were statistically analyzed.Results The extraction products of the two types of reagent kits contained 3%-9% of residual ethanol, both of which caused an impact on the amplification, potentially leading to a decrease in sensitivity. For human cytomegalovirus(HCMV), when ethanol concentration reached 4. 38%, the fluorescence gain intensity showed significant difference(P < 0. 001), and the amplification efficiency showed significant difference. For respiratory syncytial virus(RSV), there was a significant difference in fluorescence gain intensity when ethanol concentration reached 2. 63%(P = 0. 004). When the concentration of ethanol reached 3. 50%, the difference of Ct values was significant(P = 0. 004), and the amplification efficiency was significantly different.Conclusion Ethanol can affect the sensitivity, Ct values, fluorescence gain, and the amplification efficiency of qPCR. In order to detect nucleic acid with higher sensitivity, it is suggested that the residue of ethanol in the extracted products should not exceed 3. 50%.

Objective:To observe the characteristics of multi-muscle surface electromyography (sEMG) network indices during static standing among hemiplegic stroke survivors, and to evaluate the value of the indices in assessing neuromuscular dysfunction.Methods:Ten male stroke survivors with hemiplegia were recruited into the hemiplegia group, and 10 age-matched healthy males were chosen as the control group. Both groups were required to perform 30s static standing tasks with their eyes open and closed. The sEMG signals from the bilateral gluteus maximus (GM), rectus femoris (RF) and biceps femoris (BF) muscles were synchronously collected. Linear time-frequency domain indices were then calculated from the sEMG signals, including the root mean square (RMS) and median frequency (MF). Network indices were extracted from the multiplex recurrence network and weighted networks were constructed from the sEMG signals, including the average interlayer mutual information (I), average edge overlap (ω), clustering coefficient (C), average shortest path length (L) and degree of centrality (DC).Results:With the eyes closed, the RMS values of the bilateral GMs of the hemiplegia group, as well as the values for the RF and BF on the unaffected side were significantly higher than the control group′s values. In the hemiplegia group, the RMS values of the RF and BF muscles on the unaffected side were significantly higher than on the affected side during standing with the eyes closed. For the RF muscles the RMS values on the unaffected side were, on average, significantly higher than with the eyes open. The MF of the GM muscles on the unaffected side in the hemiplegia group was significantly lower than the average MF values of the bilateral GM muscles in the control group with the eyes open or closed. With the eyes closed, the MF of the unaffected-side GM was significantly lower than that of the affected-side GM in the hemiplegia group. Compared with the control group, the hemiplegia group showed a significant increase in I and ω values, but a significant decrease in L values with the eyes open or closed. The DC values of the bilateral GM, RF and BF muscles in the hemiplegia group were significantly higher than among the control group with the eyes open, which was also true of the bilateral GM and RF muscles with the eyes closed. With the BF muscles it was true only of the unaffected side. In the hemiplegia group, the DC values of the unaffected-side GM with the eyes open or closed, and of the unaffected-side BF with the eyes closed.Conclusions:When standing still, hemiplegic stroke survivors exhibit increased overall synchronous muscle adjustment with involvement of unaffected-side muscles, especially the GM. sEMG network indices such as I, ω, L and DC can assess multi-muscle synchronous adaptability and the involvement of single muscles. sEMG network algorithms thus have potential as a new method for localizing and quantitatively assessing neuromuscular dysfunction among such patients.

Objective:To investigate the clinicopathological features,treatment and prognosis of endometrial carcinoma arising in endometrial polyps(EPs).Methods:Thirty-eight cases of endometrial carcinoma arising in EPs were collected in the First Affiliated Hospital of Nanjing Medical University from January 2010 to December 2021.The clinical data,pathological characteristics,and follow-up information were retrospectively analyzed from three dimensions:tumor histological type,the menopausal status of the patients and the scope of malignant le-sions.Results:①In terms of tumor histological type,uterine endometrioid carcinoma 33 cases(86.8%)and uter-ine serous carcinoma 50 cases(13.2%),the mean age of patients with endometrioid carcinoma arising from EPs was significantly lower than that of patients with serous carcinoma arising from EPs(53.3±10.0 years vs.63.4±10.3 years,P=0.043).②In terms of menopausal status,endometrioid carcinoma progressed from atypical poly-poid adenomyoma(APA)accounted for 55.6%(10/18)and 5.0%(1/20)in the pre-and post-menopausal group,respectively(P=0.001).The rate of adjuvant therapy was significantly lower in premenopausal patients than in postmenopausal patients(16.7%vs.50.0%,P=0.043).③There were no significant differences in pa-tients age,menopausal status,BMI,comorbidities,abnormal uterine bleeding,maximum diameter of EP,tumor stage,grade,treatment or recurrence rate between cases with tumors confined and unconfined to EPs.Conclu-sions:Endometrioid and serous carcinomas progressed from EPs were more common in perimenopausal and postmenopausal patients,respectively.Malignancies derived from APA contributed to almost half of EP-arising carcinomas in premenopausal women,with a higher degree of differentiation and a low rate of postoperative adju-vant therapy.

Objective:To summarize the clinical traits, diagnostic and therapeutic experiences of pancreatic tumors in children.Methods:This is a retrospective case series study. According to inclusion and exclusion criteria, clinical data of 35 children with primary pancreatic tumors who were treated at the Department of General Surgery, the Children′s Hospital Affiliated to Xiangya School of Medicine, Central South University were collected from January 2010 to June 2023. There were 17 male and 18 female patients, aging from 3 months to 15 years and 1 month, with a median age of 9 years and 2 months. The presenting symptoms included 16 patients who had abdominal mass, 13 had clinical symptoms of abdominal pain and vomiting, 4 cases were detected on the physical examination, and 7 cases were discovered on abdominal trauma examinations.Results:Among the 35 cases, 16 cases of tumors were located in the pancreatic head. Among them, 9 cases underwent open pancreaticoduodenal surgery, 2 cases received duodenum-preserving pancreatic cephalectomy, 4 cases underwent pancreatic tumor resection, and 1 case underwent laparotomy for pancreatic tumor biopsy. Seventeen cases had tumors located in the body and tail of the pancreas, including 13 patients who underwent laparoscopic splenic-sparing caudectomy and 4 patients who underwent pancreatic tumors resection. Two cases of von Hippel-Lindau(VHL) with diffuse cystic pancreatic degeneration were continuously observed and followed up. Postoperative pathological examinations revealed that there were 20 cases of solid pseudopapillary neoplasm(SPN), 6 cases of pancreaticoblastoma(PBL), 2 cases of insulinoma, 2 cases of serous cystadenoma, 2 cases of mature teratoma and 1 case of lipoma. Four patients with PBL were completely resected after 4 cycles of induction chemotherapy due to their large tumors. All PBL patients received standardized systematic treatment including surgery and chemotherapy. Postoperative complications included 4 cases(12.1%) of biochemical leakage, 1 case(3.0%) of grade B pancreatic fistula complicated with massive bleeding, 2 cases(6.1%) of elevated platelet count, and 2 cases(6.1%) of intestinal obstruction. One case of grade B pancreatic fistula complicated with massive bleeding underwent emergency surgery, and other cases with complications were cured through conservative treatment. Two patients with SPN recurred at 36 and 33 months and were treated with reoperation. One patient with obstructive jaundice with high-grade transformed SPN was abandoned after diagnosis, and one patient with PBL died. The follow-up period of the patients was 1 to 7 years. The tumor size of 2 VHL patients with diffuse cystic pancreatic degeneration did not change after regular visits, and tumor survival was not observed in other children.Conclusions:There are no specific clinical manifestations of pancreatic tumors in children and there are various pathological types, with the majority being SPN and PBL. Radical surgery for preservation of organ and functions is the preferred treatment for pediatric pancreatic tumors, and most have a favorable prognosis.

Objective:To examine the dynamic interactive relationships between cyberbullying perpetration,anxiety,and depression in college students.Methods:A total of 4 148 college students from a comprehensive uni-versity in Guangxi Zhuang Autonomous Region completed self-report measures including Cyberbullying Scale(CS),Self-Rating Anxiety Scale(SAS),and Self-Rating Depression Scale(SDS)at two time points,18-month a-part.Results:Correlation analysis showed significant concurrent(r=0.11-0.17,Ps＜0.001)and lagged(r=0.06-0.44,Ps＜0.001)positive associations among CS,SAS,and SDS scores.Cross-lagged analysis,controlling for gender and age,indicated that T1 CS scores were positively associated with T2 SAS and SDS scores(β=0.13,0.14,Ps＜0.001).Additionally,SAS scores at T1 and SDS scores at T1 were positively associated CS scores at T2,respectively(β=0.07,0.05,Ps＜0.001).Conclusion:Cyberbullying perpetration,anxiety,and depression in-fluence each other over time among college students,suggesting a reinforcing negative cycle.

Objective To explore the current status of minor children parenting concerns among young and middle-aged breast cancer patients and investigate the influencing factors based on a random forest model so as to provide references for clinical interventions.Methods A convenience sampling method was used to select breast cancer patients undergoing treatment in our hospital between April and December 2023.A self-designed general information questionnaire,the Chinese version of parenting concerns questionnaire(PCQ),perceived social support scale(PSSS),concern about recurrence scale(CARS),and the brief illness perception questionnaire(BIPQ)were used for the study.A random forest model and the least absolute shrinkage and selection operator(LASSO)were employed to prioritise variables and filtered by significance.The selected variables were then incorporated into the multiple linear regression analysis.Results A total of 260 patients completed the study.The score of minor children parenting concerns of young and middle-aged breast cancer patients was 51.1±6.4.The multiple linear regression analysis,which included variables determined by random forest and LASSO regression(and sorted by the importance of influencing factors),showed that higher disease perception,lower perceived social support,greater concern about cancer recurrence,stage IV tumors,being divorced/widowed,and having more minor children were associated with higher parenting concerns among young and middle-aged breast cancer patients(all P<0.05),accounting for 57.0%of the total variance.Conclusion The minor children parenting concerns in young and middle-aged breast cancer patients are at a moderately high level and are influenced by a variety of factors.Healthcare professionals should develop targeted measures and interventions to reduce the parenting concerns among the patients.

Objective:To investigate the clinical characteristics, diagnosis and treatment of duodenal duplication in children.Methods:From Jan 2017 to Jun 2023, clinical data of 8 child patients complaining nonspecific intraabdominal symptoms at Department of General Surgery, Hu'nan Children's Hospital undering surgery were retrospectively analyzed.Results:Five patients had duodenal duplication resected, 3 patients underwent fenestration and mucous membraine removel.Postoperative pathology showed cyst type in 6 cases, tubular type in 2 cases,all with lining of intestinal mucosa, and smooth muscle, including 1 case with ectopic pancreas tissues, 2 cases with ectopic gastric mucosa tissues. Postoperative follow-up were 10-48 months. During the follow-up period, there was no abdominal pain, vomiting, abdominal distension, and other symptoms, and the children recovered well.Conclusions:The clinical manifestations of duodenal duplication are no specific. Definite diagnosis depends on pathology. Duodenal duplication resection or fenestration plus mucosal stripping is the treatment of choice. The prognosis is good.

Objective:To screen differentially expressed genes (DEGs) associated with liver cancer by bioinformatics analysis method, and to investigate the mechanism of the minichromosome maintenance protein 2 ( MCM2) and replication factor C subunit 4 ( RFC4) genes in liver cancer in vitro. Methods:Gene expression profiling data of 80 and 36 hepatocellular carcinoma tissues and 307 and 13 cirrhotic tissues were obtained from the GSE25097 and GSE98620 datasets of the gene expression analysis (GEO) database, respectively. Gene expression profiling data of 374 liver cancer tissues and 50 normal liver tissues were downloaded from the cancer genome atlas (TCGA) database. Limma and DESeqs R software were used to process the gene expression profiling data, construct protein-protein interaction networks, and analysis the relevance of these genes to survival. Weighted gene co-expression network analysis was performed to screen out the core genes. Liver cancer SMMC7721 cells were transfected with MCM2 blank plasmid (MCM2 control group), MCM2 overexpression plasmid [MCM2 WT1 group, MCM2 WT2 (2-fold WT1) group], RFC4 blank plasmid (RFC4 control group), and RFC4 overexpression plasmid [RFC4 WT1 group, RFC4 WT2 (2-fold WT1) group], respectively. The expression of MCM2 and RFC4 in liver cancer cell lines and their transfection levels were detected by real-time fluorescence quantitative reverse transcription PCR and Western blotting. The effects of MCM2 and RFC4 on the proliferation of hepatocellular carcinoma cells were detected by MTT assay and cell cloning assay, respectively. The effects of MCM2 and RFC4 on the migration of liver cancer cells were determined by the scratch assay. The effects of MCM2 and RFC4 on liver cancer cell invasion were detected by Transwell assay.Results:By bioinformatic analysis, 9 HCC DEGs were selected, including ubiquitin conjugating enzyme E2 T ( UBE2T), aurora kinase A ( AURKA), targeting protein for Xklp2 ( TPX2), MCM2, RFC4, ribonucleoside-diphosphate reductase subunit M2 ( RRM2), serine peptidase inhibitory factor Kazal type 1 ( SPINK1), collagen type XV alpha 1 chain ( COL15A1) and C-C motif chemokine 25 ( CCL25). Among the six genes associated with clinical stages, the MCM2 and RFC4 genes were found to be strongly associated with prognosis in liver cancer. The relative protein expression of MCM2 and RFC4 in HepG2 cells (1.83±0.07, 1.44±0.09) and SMMC7721 cells (1.74±0.05, 1.43±0.08) was higher than that in MIHA cells (1.00±0.02, 1.00±0.03), and all the differences were statistically significant (all P<0.05). The relative gene expression of MCM2 and RFC4 in HepG2 cells (14.30±0.12, 5.10±0.18) and SMMC7721 cells (10.60±0.11, 7.60±0.07) was higher than that in MIHA cells (1.00±0.05, 1.00±0.03), and all the differences were statistically significant (all P<0.05). Compared with the MCM2 control group, the absorbance values [(0.28±0.01 and 0.21±0.01) vs 0.18±0.03], the number of clonal cells [(717±12 and 782±29) cells vs (389±17) cells], the percentage migration [(0.43±0.02 and 0.68±0.01) vs 0.15±0.06], and the number of cellular invasions [(933±21 and 821±11) cells vs (409±16) cells] were higher in the MCM2 WT1 and MCM2 WT2 groups, and the differences were all statistically significant (all P<0.05). Compared with the RFC4 control group, the absorbance values [(0.30±0.02 and 0.21±0.01) vs 0.17±0.02], the number of cloned cells [(571±11 and 728±9) cells vs (373±23) cells], the percentage migration [(0.75±0.11 and 0.67±0.04) vs 0.34±0.07], and the number of cell invasion [(835±26 and 818±18) cells vs (629±12) cells] were higher in the RFC4 WT1 and the RFC4 WT2 groups, and the differences were statistically significant (all P<0.05). Conclusions:MCM2 and R FC4 genes play a role in promoting tumorigenesis and growth in hepatocellular carcinoma.

Objective:To explore the clinicopathological characteristics and prognosis of fumarate hydratase-deficient uterine leiomyoma (FH-dUL).Methods:Clinical data and follow-up information for 117 patients with FH-dUL diagnosed through surgical pathology and immunohistochemistry in the First Affiliated Hospital of Nanjing Medical University from January 2020 to December 2024, were collected. A control group of 130 patients with common uterine leiomyomas was also included. The differences between the two groups in clinical, imaging, and pathological characteristics were compared. Additionally, recurrence rates, fertility outcomes for FH-dUL patients, and the incidence of renal cancer in FH germline mutation carriers were monitored.Results:(1) Comparison of clinicopathological characteristics: the median age of 117 FH-dUL patients was 35 years, and the median age at first diagnosis of uterine leiomyomas was 29 years, both significantly younger than the control group (41 and 36 years; both P<0.01). The FH-dUL group showed significantly higher incidences of uterine myomectomy, multiple leiomyomas, diffusion restriction on pelvic magnetic resonance imaging diffusion weighted imaging, and typical pathological features (candelabra-like vessels, bizarre nuclei, cytoplasmic eosinophilic globules, perinuclear halo, cellular atypia) and higher ultrasound blood flow score (all P<0.05). Of the 30 FH-dUL patients who underwent genetic testing, 9 had germline mutations, 3 had somatic mutations, and 6 had mutations of unclear origin. Among the 9 FH gene germline mutation patients, 2 had already developed renal cell carcinoma. (2) Recurrence analysis: among the 56 patients who underwent uterine myomectomy, 22 (39.3%, 22/56) experienced recurrence during follow-up, compared to 12 (21.8%, 12/55) of the 55 patients in the control group, the difference between the two groups was statistically significant ( P=0.046). Multivariate binary logistic regression analysis showed that cellular leiomyomas ( OR=9.489, 95% CI: 1.740-51.755; P=0.009) and multiple uterine leiomyomas ( OR=10.709, 95% CI: 1.354-84.683; P=0.025) were significant risk factors for recurrence in FH-dUL. (3) Fertility analysis: among the 66 FH-dUL patients who underwent fertility-preserving surgery, 16 had the intention to have fertility desire, only 2 (2/16) completed their fertility plans during follow-up. Conclusions:Clinicopathological features and imaging features help to differentiate FH-dUL from common type uterine fibroids, but lack specificity, and the diagnosis of FH-dUL is based on immunohistochemistry. The recurrence rate after resection of FH-dUL is high, and cellular and multiple leiomyomas are important predictors of recurrence. It is crucial to perform genetic testing, genetic counseling, drug treatment to prevent recurrence, fertility guidance, and long-term comprehensive management after surgery for FH-dUL management.

Objective:To study the strategies in the diagnosis and treatment of solid pseudopapillary neoplasm (SPN) of the pancreas in children.Methods:The clinical data of 16 children with SPN managed at the Affiliated Children's Hospital of Xiangya School of Medicine, Central South University (Hunan Children's Hospital) from January 2015 to December 2022 were retrospectively studied. There were 9 males and 7 females, ranged from 7 years and 5 months to 13 years and 3 months, with a median age of 10 years and 7 months. The clinical performance and types of the tumor (type Ⅰ, tumor located in the tail of pancreas; Type Ⅱ, the tumor located in the pancreatic body; Type Ⅲ, tumor located in the head of the pancreas), surgical methods, complications (pancreatic fistula, intestinal obstruction, hemorrhage) were analyzed. The recurrence was followed up by outpatient review or telephone after surgery.Results:The main clinical characteristics included abdominal pain (12 cases), abdominal trauma examination findings (2 cases), and physical examination findings (2 cases). The results of abdominal ultrasound, enhanced CT and/or MRI examinations showed that 4 cases of cystic tumors, 7 cases of solid tumors, and 5 cases of cystic solid tumors. Among of all patients, 2 cases were clinical classification type Ⅰ, 8 cases were type Ⅱ, and 6 cases were type Ⅲ. Among 6 cases with tumor located in the head of the pancreas, 3 patients underwent pancreaticoduodenectomy, 1 patient underwent resection of the head of the pancreas with preservation of the duodenum, 1 patient underwent laparoscopic resection of pancreatic head tumor, 1 patient underwent biopsy of pancreatic head tumor. The other 10 cases with tumor located in the body and tail of the pancreas, 6 patients underwent laparoscopic splenic preservation pancreatic body-tail resection, 1 patient underwent laparoscopic resection of pancreatic body-tail and splenectomy, combined with autologous splenic transplantation, 1 patient underwent laparoscopic removal of the splenic venous thrombus, splenic preservation pancreatic body-tail, 1 patient underwent laparoscopic enucleation of a dorsal tumor in the middle part of the pancreas, and 1 patient underwent open resection of spleen-preserving pancreatic body-caudal. Postoperative complications included pancreatic fistula (grade B) with massive hemorrhage (1 case), biochemical leakage (1 case), post-operative pancreatitis (1 case), intestinal obstruction (1 case), and reactive thrombocytosis (1 case). The patient pancreatic fistula (grade B) with massive hemorrhage and pancreatic leakage required emergency surgery, while the remaining patients recovered with conservative treatment. All patients were followed up for 22 to 90 months (median follow-up period of 70 months), two cases relapsed and were successfully re-treated surgically at 36 months and 33 months post-surgery, respectively. One patient with high grade transformation SPN and obstructive jaundice withdrew treatment (missed follow-up), while the remaining patients without recurrence after surgery.Conclusion:Children SPN has no specific clinical features, and preoperative abdominal ultrasound, CT and/or MRI can be located and diagnosed. Surgery is the preferred treatment, and the prognosis is well.