Genetic transformation is a fundamental tool in molecular biology research of medicinal plants. Tailoring transgenic technologies to each distinct medicinal plant would necessitate a substantial investment of time and effort. Here, we present a simple hairy root transformation method that does not require sterile conditions, utilizing Agrobacterium rhizogenes strain K599 and the visible RUBY reporter system. Transgenic hairy roots were obtained for six tested medicinal plant species, roots or rhizomes of which have recognized medicinal value, spanning four botanical families and six genera (Platycodon grandiflorus, Atractylodes macrocephala, Scutellaria baicalensis, Codonopsis pilosula, Astragalus membranaceus, and Glycyrrhiza uralensis). Furthermore, two previously identified Glycyrrhiza uralensis UGTs that convert liquiritigenin into liquiritin in heterologous systems were studied in planta using the method. Our results indicate that overexpression of GuUGT1 but not GuUGT10 and Cas9-mediated knockout of GuUGT1 profoundly influenced the accumulation of liquiritin and isoliquiritin in licorice roots. Therefore, the method described here represents a simple, rapid and widely applicable hairy root transformation method that enables fast gene functional study in medicinal plants.

Objective To analyze the predictive value of spermatic vein diameter in diagnosing occult oblique inguinal hernia in children.Methods A total of 93 children of oblique inguinal hernia were admitted to Hangzhou Linping District Maternal &Child Health Care Hospital from August 2021 to August 2023 as research objects,including 15 with 30 sides of bilateral dominant hernia and 78 with 78 sides of unilateral dominant hernia.Among them,21 with 21 sides of contralateral recessive hernia were found by laparoscopic exploration in unilateral dominant hernia.The 108 sides of dominant hernia were used as dominant hernia group and 21 sides of occult hernia group as recessive hernia group.According to different ages,the patients were divided into 1 to 3 years old group 60 cases,4 to 6 years old group 33 cases;Children with oblique inguinal hernia were divided into＜2 years group(68 cases)and ≥2 years group(25 cases).The color ultrasound diagnostic instrument uses a 7.5MHz linear transducer to measure the diameter of the spermatic vein in the inguinal region of the child using blood flow imaging with a signal displayed at 2mm/s.Receiver operating characteristic(ROC)curve analysis of the predictive value of spermatic vein diameter for the diagnosis of occult inguinal hernia in children.Results Compared with occult hernia group,diameter of spermatic cord was increased in dominant hernia group(P＜0.05).Compared with overt hernia group,diameter of spermatic cord in＜2 year group and ≥2 year group was smaller,and difference was statistically significant(P＜0.05).In prediction of disease diagnosis with different age and course of disease,sensitivity of 1 to 3 years old was lower than that of 4 to 6 years old,and specificity was higher.The sensitivity of ≥2 years group was lower than that of＜2 years group.The specificity was higher and difference was statistically significant(P＜0.05).Conclusion diameter of spermatic vein can be used to predict the diagnosis of occult oblique inguinal hernia in children with high sensitivity and specificity.

Objective:To construct a prediction model for postoperative poor in-hospital prognosis in patients with traumatic brain injury (TBI) and evaluate its predictive performance.Methods:A retrospective case control study was conducted to analyze the clinical data of 1 120 TBI patients admitted to Changde Hospital Affiliated to Xiangya Medical College of Central South University from May 2019 to December 2024. The patients were divided into the training set ( n=784) and verification set ( n=336) at a ratio of 7∶3. Based on the Glasgow outcome scale-extended (GOS-E) at discharge, the training set was stratified into favorable prognosis group ( n=335, GOS-E 5-8 points) and poor prognosis group ( n=449, GOS-E 1-4 points). The two groups in the training set were compared in terms of general baseline indicators, TBI-related clinical indicators, and admission laboratory blood test results. Univariate analysis and Lasso regression analysis were employed to screen risk factors associated with postoperative poor in-hospital prognosis in TBI patients. Multivariate Logistic regression analysis was used to determine independent risk factors and construct a regression equation. The regression equation was presented using R language to create a visual nomogram for predicting postoperative poor in-hospital prognosis in TBI patients. In both the training set and verification set, the predictive performance of the model was evaluated by calculating the area under the receiver operating characteristic (ROC) curve (AUC), plotting calibration curves, and performing decision curve analysis (DCA). Results:The results of the univariate analysis indicated that the age, Charlson complication index (CCI), time from trauma to admission, time from trauma to operation, cause of injury, abbreviated injury scale (AIS) (head and neck), injury severity score (ISS), admission Glasgow coma scale (GCS), admission pupil responsiveness, multiple craniocerebral injuries, subdural hematoma, intracerebral hematoma, intraventricular hemorrhage, subarachnoid hemorrhage, decompressive craniotomy, intraoperative blood loss, intraoperative blood transfusion, traumatic cerebral infarction, postoperative delayed bleeding, epilepsy seizures, as well as the following admission tested results including red blood cell count, white blood cell count, platelet count, neutrophil percentage, percentage of lymphocytes, albumin, total bilirubin, urea nitrogen, thrombin time (TT), prothrombin time (PT), international standardized ratio (INR), glutamic aminotransferase, alanine aminotransferase, creatinine, and blood glucose were statistically different between the two groups in the training set ( P<0.05). Lasso regression analysis suggested 14 risk factors of age, CCI, cause of injury, head and neck AIS, ISS, admission GCS, admission pupil responsiveness, multiple craniocerebral injuries, subdural hematoma, intracerebral hematoma, intraoperative blood loss, admission platelet count, admission albumin, admission blood glucose for postoperative poor in-hospital prognosis. The results of the multivariate Logistic regression analysis showed that age ( OR=1.02, 95% CI 1.00, 1.03, P<0.01), CCI ( OR=1.46, 95% CI 1.02, 2.09, P<0.05), head and neck AIS ( OR=1.43, 95% CI 1.11, 1.85, P<0.01), ISS ( OR=2.16, 95% CI 1.39, 3.35, P<0.01), admission GCS ( OR=1.59, 95% CI 1.19, 2.13, P<0.01), intracerebral hematoma ( OR=4.41, 95% CI 2.15, 9.44, P<0.01), intraoperative blood loss ( OR=1.05, 95% CI 1.00, 1.09, P<0.05), admission platelet count ( OR=0.98, 95% CI 0.97, 0.99, P<0.01), admission blood glucose ( OR=1.08, 95% CI 1.02, 1.15, P<0.05) could be the main risk factors to construct a prediction model for postoperative poor in-hospital prognosis in TBI patients. Meanwhile, a regression equation was constructed: Logit[ P/(1- P)]=-2.4+ 0.02×"age"+0.38×"CCI"+0.36×"head and neck AIS"+0.77×"ISS"+0.47×"admission GCS"+1.48×"intracerebral hematoma"+0.05×intraoperative blood loss-0.02×admission platelet count+0.08×admission blood glucose. In the training set, the predictive model for poor postoperative in-hospital prognosis in TBI patients achieved an AUC of 0.87 (95% CI 0.84, 0.89), with a Youden′s index of 0.57, sensitivity of 73.70%, and specificity of 83.00%. In the verification set, the model showed an AUC of 0.80 (95% CI 0.76, 0.85), with a Youden′s index of 0.63, sensitivity of 65.20%, and specificity of 77.90%. In the training set, the Brier score for the calibration curve was 0.14 (95% CI 0.13, 0.16). In the verification set, the Brier score for the calibration curve was 0.18 (95% CI 0.15, 0.20). The DCA diagram indicated that the nomogram prediction model provided high clinical net benefit for predicting postoperative poor in-hospital prognosis in TBI patients. Conclusion:The prediction model for postoperative poor in-hospital prognosis in TBI patients, constructed based on age, CCI, head and neck AIS, ISS, admission GCS, intracerebral hematoma, intraoperative blood loss, admission platelet count, and admission blood glucose, exhibits good predictive performance.

Background and aims:Early and accurate diagnosis of the coexistence of Wilson's disease(WD)and chronic hepatitis B(CHB)presents a significant challenge for clinicians.The objective of this study was to retrospectively analyse the characteristics of such patients to improve clinical practice and provide a reference for clinical management.Methods:From January 2011 to December 2022,35 patients with concurrent CHB and WD(CHB+WD group)were identified.A total of 127 patients with CHB(CHB group)and 168 patients with WD(WD group)were included in the control group between January 2016 and December 2021.Propensity score matching(PSM)was performed to balance the baseline values between groups.The Kaplan-Meier(K-M)survival analysis and log-rank test were performed to compare the prognoses.Results:In the cohort of 35 patients with concurrent CHB and WD,74.3％of patients(26 patients)faced a substantial delay of up to 10 years(range:0-40 years)in WD diagnosis following their CHB diagnosis.Twenty-three(65.7％)patients had cirrhosis at the time of WD diagnosis,and 26(74.3％)patients experienced liver failure.The levels of serum copper and uric acid were lower in patients in the CHB+WD group than in those in the CHB group.Patients in the CHB+WD group presented higher alanine transaminase and total bile acid levels compared to those in the WD group.K-M survival analysis indicated that patients with CHB and WD had poorer outcomes than those with CHB alone;however,the outcomes were similar to those of individuals with WD alone.The optimal cut-point of serum ceruloplasmin(CP)in identifying WD in CHB patients was 0.10 g/L before PSM and after PSM.Conclusions:The present study emphasizes the importance of clinicians being vigilant for concurrent CHB and WD diagnoses,as delays in WD diagnosis may adversely affect patient outcomes.CHB patients with serum CP below 0.10 g/L are highly recommended to screen for WD.

Objective To investigate the relationships of cerebrospinal fluid levels of collagen triple helix repeat containing-1(CTHRC1)and olfactomedin-3(OLFM3)with cognitive impairment and cere-brospinal fluid biomarker levels in patients with Alzheimer's disease(AD).Methods Ninety-six patients with AD were selected as study objects(AD group),and divided into mild group(n=34),moderate group(n=39)and severe group(n=33)according to Clinical Deentia Scale(CDR)score.Sixty patients without cognitive impairment who underwent lumbar puncture during the same period served as the control group.Enzyme-linked immunosorbent assays were used to measure cere-brospinal fluid levels of CTHRC1,OLFM3 and biomarkers[β-amyloid(Aβ)-40,Aβ-42,Aβ42/Aβ-40,total tau(T-tau)and phosphorylated tau(P-tau)].Cognitive impairment in AD patients was as-sessed using the Mini-Mental State Examination(MMSE)and Montreal Cognitive Assessment(MoCA).The relationships of CSF CTHRC1 and OLFM3 levels with cognitive impairment and cerebrospinal flu-id biomarkers were analyzed.Results The AD group showed significantly higher cerebrospinal fluid levels of CTHRC1,T-tau and P-tau,and significantly lower levels of OLFM3,Aβ-42 and Aβ42/Aβ-40 compared to the control group(P＜0.05).The area under the curve(AUC)for diagnosing AD with cerebrospinal fluid CTHRC1 and OLFM3 was 0.839 and 0.822,respectively,and the com-bined AUC was 0.923.Cerebrospinal fluid CTHRC1 of mild group,moderate group and severe group were increased successively,and OLFM3 was decreased successively,the difference was sta-tistically significant(P＜0.05).Cerebrospinal fluid Aβ-42 and Aβ-42/Aβ-40 in mild,moderate and severe groups were decreased successively,while T-tau and P-tau were increased successively,with statistical significance(P＜0.05).The MMSE and MoCA scores of mild group,moderate group and severe group decreased successively,and the difference was statistically significant(P＜0.05).Cerebrospinal fluid CTHRC1 levels were positively correlated with disease severity,while OLFM3 levels were negatively correlated(P＜0.05).Cerebrospinal fluid CTHRC1 was negatively correlated with Aβ42,Aβ42/Aβ-40,MMSE scores and MoCA scores,and positively correlated with T-tau and P-tau(P＜0.05).Cerebrospinal fluid OLFM3 was positively correlated with Aβ-42,Aβ42/Aβ-40,MMSE scores and MoCA scores,and negatively correlated with T-tau and P-tau(P＜0.05).Conclusion In the cerebrospinal fluid of patients with AD,CTHRC1 is elevated while OLFM3 is decreased.Both CTHRC1 and OLFM3 are associated with the severity of AD,cog-nitive impairment and levels of cerebrospinal fluid biomarkers.

Objective:To reveal the differences in the transcriptome maps of brain tissues in mice subjected to Flash irradiation and conventional dose rate irradiation with electron beams and to explain the biological effect and mechanisms of Flash irradiation from multiple perspectives.Methods:Following the principle of grouping based on approximate body weights, 36 female C57BL/6J mice were divided into three groups, i. e., the control, conventional dose rate irradiation (CONV), and Flash irradiation (Flash) groups, with 12 mice in each group. Both the CONV and Flash groups received a single 15 Gy whole-brain irradiation with 9 MeV electron beams. At 3 d post-irradiation, the whole-brain tissue specimens were collected for hematoxylin-eosin (HE) staining to observe pathological changes. At 1, 3, and 10 weeks post-irradiation, the motion function, cognitive ability, depression level, and spatial memory capacity of the mice were assessed using ethology. At 1 and 10 weeks after behavioral experiments, brain tissue samples were collected and snap-frozen in liquid nitrogen for reference-based transcriptome sequencing. Accordingly, the differences in the transcriptome maps of radiation-induced brain injury between CONV and Flash groups were analyzed.Results:The HE staining-based pathological result revealed that compared to the CONV group, the Flash group exhibited reduced glial cell hyperplasia and inflammatory cell infiltration in brain tissues. Ethological research result at 1 week post-irradiation showed that the CONV group manifested a significantly decreased total traveled distance compared to the control and Flash groups ( t = 5.51, 2.38, P < 0.05) and a significantly increased immobility time compared to the control group ( t = 3.60, P < 0.05). Ethological research result at 3 weeks post-irradiation indicated that compared to the CONV group, the Flash group displayed significantly alleviated cognitive impairment ( t = 3.35, P < 0.05) and reduced depression levels ( t = 2.39, P < 0.05). Ethological research result at 10 weeks post-irradiation demonstrated that the CONV group showed the worst cognitive performance, significantly differing from the control group ( t = 4.53, P < 0.05). Transcriptome sequencing result revealed that besides immune-related pathways, the Flash group also exhibited multiple upregulated metabolic pathways and fibroblast growth factor (FGF)-related pathways compared to the CONV group. Conclusions:Compared to conventional dose rate irradiation, Flash irradiation can effectively alleviate radiation-induced brain injury in mice. This effect is associated with various metabolic pathways (including amino acid metabolism) and FGF-related pathways besides immune pathways.

Objective:To discuss the feasibility and safety of stage I anastomosis and T-tube fistulation in laparoscopic local resection of duodenal tumors.Methods:A descriptive case series study was used to retrospectively analyze the clinical diagnosis and treatment data of 14 patients with duodenal tumors who successfully underwent laparoscopic local resection of duodenal tumors + phase I anastomosis + T-tube ostomy in the Guangdong Provincial Hospital of Chinese Medicine and the First Affiliated Hospital of Guangzhou University of Chinese Medicine from October 2021 to March 2024. The resection and reconstruction steps of laparoscopic local resection of duodenal tumor + phase I anastomosis + T-tube ostomy are as follows: (1) after the safe margin is clear, the duodenal tumor is completely removed in full thickness, and the specimen bag is taken out and sent to frozen section to determine the nature of the tumor and the negative margin; (2) Perforate the anterior duodenal wall below the tumor plane, place a 16# T tube, and fix it with laparoscopic purse string suture. The abdominal wall is led out through the duodenum, and the duodenal T tube fistulation is performed; (3) The duodenum was continuously sutured in a full-thickness transverse shape, and the seromuscular layer was strengthened to form a phase I anastomosis. The nutritional improvement of patients after operation was mainly observed, and the intraoperative situation and postoperative complications were recorded.Results:No conversion to laparotomy, postoperative emergency reoperation, intraoperative and postoperative complications occurred in 14 patients with duodenal tumors who completed laparoscopic local resection of duodenal tumors + phase I anastomosis + T-tube ostomy. The operation time was (225.43 ± 56.54) min, and the intraoperative blood loss was (72.14 ± 74.65) ml. The patient recovered well after operation, and no severe postoperative abdominal bleeding occurred. Postoperative gastrointestinal angiography showed that the anastomotic stoma was unobstructed, and there were no stenosis, anastomotic leakage and other related complications. There was no significant difference in serum albumin [(37.09 ± 3.53) g/L vs. (37.52 ± 4) g/L] and hemoglobin [(100.79 ± 31.93) g/L vs. (103.07 ± 19.6) g/L] between before and 1 week after operation ( P > 0.05). Conclusion:Laparoscopic local resection of duodenal tumor + phase I anastomosis + T-tube fistulation can be used as one of the safe and feasible improved methods for local resection of duodenal tumor to effectively reduce the occurrence of related complications.

Objective:To reveal the differences in the transcriptome maps of brain tissues in mice subjected to Flash irradiation and conventional dose rate irradiation with electron beams and to explain the biological effect and mechanisms of Flash irradiation from multiple perspectives.Methods:Following the principle of grouping based on approximate body weights, 36 female C57BL/6J mice were divided into three groups, i. e., the control, conventional dose rate irradiation (CONV), and Flash irradiation (Flash) groups, with 12 mice in each group. Both the CONV and Flash groups received a single 15 Gy whole-brain irradiation with 9 MeV electron beams. At 3 d post-irradiation, the whole-brain tissue specimens were collected for hematoxylin-eosin (HE) staining to observe pathological changes. At 1, 3, and 10 weeks post-irradiation, the motion function, cognitive ability, depression level, and spatial memory capacity of the mice were assessed using ethology. At 1 and 10 weeks after behavioral experiments, brain tissue samples were collected and snap-frozen in liquid nitrogen for reference-based transcriptome sequencing. Accordingly, the differences in the transcriptome maps of radiation-induced brain injury between CONV and Flash groups were analyzed.Results:The HE staining-based pathological result revealed that compared to the CONV group, the Flash group exhibited reduced glial cell hyperplasia and inflammatory cell infiltration in brain tissues. Ethological research result at 1 week post-irradiation showed that the CONV group manifested a significantly decreased total traveled distance compared to the control and Flash groups ( t = 5.51, 2.38, P < 0.05) and a significantly increased immobility time compared to the control group ( t = 3.60, P < 0.05). Ethological research result at 3 weeks post-irradiation indicated that compared to the CONV group, the Flash group displayed significantly alleviated cognitive impairment ( t = 3.35, P < 0.05) and reduced depression levels ( t = 2.39, P < 0.05). Ethological research result at 10 weeks post-irradiation demonstrated that the CONV group showed the worst cognitive performance, significantly differing from the control group ( t = 4.53, P < 0.05). Transcriptome sequencing result revealed that besides immune-related pathways, the Flash group also exhibited multiple upregulated metabolic pathways and fibroblast growth factor (FGF)-related pathways compared to the CONV group. Conclusions:Compared to conventional dose rate irradiation, Flash irradiation can effectively alleviate radiation-induced brain injury in mice. This effect is associated with various metabolic pathways (including amino acid metabolism) and FGF-related pathways besides immune pathways.

Objective:To discuss the feasibility and safety of stage I anastomosis and T-tube fistulation in laparoscopic local resection of duodenal tumors.Methods:A descriptive case series study was used to retrospectively analyze the clinical diagnosis and treatment data of 14 patients with duodenal tumors who successfully underwent laparoscopic local resection of duodenal tumors + phase I anastomosis + T-tube ostomy in the Guangdong Provincial Hospital of Chinese Medicine and the First Affiliated Hospital of Guangzhou University of Chinese Medicine from October 2021 to March 2024. The resection and reconstruction steps of laparoscopic local resection of duodenal tumor + phase I anastomosis + T-tube ostomy are as follows: (1) after the safe margin is clear, the duodenal tumor is completely removed in full thickness, and the specimen bag is taken out and sent to frozen section to determine the nature of the tumor and the negative margin; (2) Perforate the anterior duodenal wall below the tumor plane, place a 16# T tube, and fix it with laparoscopic purse string suture. The abdominal wall is led out through the duodenum, and the duodenal T tube fistulation is performed; (3) The duodenum was continuously sutured in a full-thickness transverse shape, and the seromuscular layer was strengthened to form a phase I anastomosis. The nutritional improvement of patients after operation was mainly observed, and the intraoperative situation and postoperative complications were recorded.Results:No conversion to laparotomy, postoperative emergency reoperation, intraoperative and postoperative complications occurred in 14 patients with duodenal tumors who completed laparoscopic local resection of duodenal tumors + phase I anastomosis + T-tube ostomy. The operation time was (225.43 ± 56.54) min, and the intraoperative blood loss was (72.14 ± 74.65) ml. The patient recovered well after operation, and no severe postoperative abdominal bleeding occurred. Postoperative gastrointestinal angiography showed that the anastomotic stoma was unobstructed, and there were no stenosis, anastomotic leakage and other related complications. There was no significant difference in serum albumin [(37.09 ± 3.53) g/L vs. (37.52 ± 4) g/L] and hemoglobin [(100.79 ± 31.93) g/L vs. (103.07 ± 19.6) g/L] between before and 1 week after operation ( P > 0.05). Conclusion:Laparoscopic local resection of duodenal tumor + phase I anastomosis + T-tube fistulation can be used as one of the safe and feasible improved methods for local resection of duodenal tumor to effectively reduce the occurrence of related complications.

Objective To analyze the predictive value of spermatic vein diameter in diagnosing occult oblique inguinal hernia in children.Methods A total of 93 children of oblique inguinal hernia were admitted to Hangzhou Linping District Maternal &Child Health Care Hospital from August 2021 to August 2023 as research objects,including 15 with 30 sides of bilateral dominant hernia and 78 with 78 sides of unilateral dominant hernia.Among them,21 with 21 sides of contralateral recessive hernia were found by laparoscopic exploration in unilateral dominant hernia.The 108 sides of dominant hernia were used as dominant hernia group and 21 sides of occult hernia group as recessive hernia group.According to different ages,the patients were divided into 1 to 3 years old group 60 cases,4 to 6 years old group 33 cases;Children with oblique inguinal hernia were divided into＜2 years group(68 cases)and ≥2 years group(25 cases).The color ultrasound diagnostic instrument uses a 7.5MHz linear transducer to measure the diameter of the spermatic vein in the inguinal region of the child using blood flow imaging with a signal displayed at 2mm/s.Receiver operating characteristic(ROC)curve analysis of the predictive value of spermatic vein diameter for the diagnosis of occult inguinal hernia in children.Results Compared with occult hernia group,diameter of spermatic cord was increased in dominant hernia group(P＜0.05).Compared with overt hernia group,diameter of spermatic cord in＜2 year group and ≥2 year group was smaller,and difference was statistically significant(P＜0.05).In prediction of disease diagnosis with different age and course of disease,sensitivity of 1 to 3 years old was lower than that of 4 to 6 years old,and specificity was higher.The sensitivity of ≥2 years group was lower than that of＜2 years group.The specificity was higher and difference was statistically significant(P＜0.05).Conclusion diameter of spermatic vein can be used to predict the diagnosis of occult oblique inguinal hernia in children with high sensitivity and specificity.