Objective To investigate the effects of palmatine on migration,invasion and epithelial mesenchymal transformation(EMT)in human oral cancer KB cells.Methods KB cells were divided into control group and palmatine-L,-M,-H groups,cultured with 0,4,8 and 16 μmol·L-1 palmatine.After incubation for 48 h,scratch assay was used to detect migration;Traswell assay was used to detect invasion;matrix metalloproteinase 2(MMP-2),MMP-9 and fibronectin(FN)contents were detected by enzyme-linked immunosorbent assay;the expression of Vimentin,N-cadherin and E-cadherin mRNA were detected by real-time quantitative polymerase chain reaction;the expression of Vimentin,N-cadherin,E-cadherin,Wnt3 and β-catenin protein were detected by Western blot.Results Cell mobility in control group and palmatine-L,-M,-H groups were(69.27±8.62)％,(52.94±4.49)％,(45.22±5.05)％and(37.63±4.88)％;the number of transmembrane cells were 197.33±20.26,125.33±12.01,97.00±9.17 and 62.67±7.51;the content of MMP-2 were(2.93±0.21),(1.49±0.13),(1.16±0.15)and(0.95±0.09)ng·mL-1;the content of MMP-9 were(3.51±0.36),(2.37±0.23),(2.06±0.35)and(1.72±0.16)ng·mL-1;the content of FN were(41.28±4.02),(24.03±3.17),(20.67±2.63)and(13.82±2.19)ng·mL-1;the above indexes in palmatine-L,-M,-H groups were compared with the control group,and the differences were statistically significant(P＜0.05,P＜0.01).The mRNA and protein expressions of Vimentin,N-cadherin and E-cadherin,and the expressions of Wnt3 and β-catenin protein in palmatine-L,-M,-H groups were statistically significant compared with those in control group(P＜0.05,P＜0.01).Conclusion Palmatine can inhibit the migration,invasion and EMT of human oral cancer KB cells,and its mechanism is related to the regulation of Wnt/β-catenin signaling pathway.

ObjectiveHuman Ku70 protein mainly involves the non-homologous end joining (NHEJ) repair of double-stranded DNA breaks (DSB) through its DNA-binding properties, and it is recently reported having an RNA-binding ability. This paper is to explore whether Ku70 has RNA helicase activity and affects miRNA maturation. MethodsRNAs bound to Ku protein were analyzed by RNA immunoprecipitation sequencing (RIP-seq) and bioinfomatic anaylsis. The expression relationship between Ku protein and miRNAs was verified by Western blot (WB) and quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) assays. Binding ability of Ku protein to the RNAs was tested by biolayer interferometry (BLI) assay. RNA helicase activity of Ku protein was identified with EMSA assay. The effect of Ku70 regulated miR-124 on neuronal differentiation was performed by morphology analysis, WB and immunofluorescence assays with or without Zika virus (ZIKV) infection. ResultsWe revealed that the Ku70 protein had RNA helicase activity and affected miRNA maturation. Deficiency of Ku70 led to the up-regulation of a large number of mature miRNAs, especially neuronal specific miRNAs like miR-124. The knockdown of Ku70 promoted neuronal differentiation in human neural progenitor cells (hNPCs) and SH-SY5Y cells by boosting miR-124 maturation. Importantly, ZIKV infection reduced the expression of Ku70 whereas increased expression of miR-124 in hNPCs, and led to morphologically neuronal differentiation. ConclusionOur study revealed a novel function of Ku70 as an RNA helicase and regulating miRNA maturation. The reduced expression of Ku70 with ZIKV infection increased the expression of miR-124 and led to the premature differentiation of embryonic neural progenitor cells, which might be one of the causes of microcephaly.

Objective To establish a nomogram model for efficiently predicting overall survival(OS)and cancer-specific survival(CSS)in patients with CRCHBM.Method 2239 patients from 2010 to 2019 were retrospectively analyzed from the Surveillance,Epidemiology,and End Results Program(SEER)databases and Wuhan Union Hospital Cancer Center.SEER is randomly assigned to the training and internal validation cohorts,and the Wuhan database serves as the external validation.Cox regression analyses were used to determine the independent clinicopathological prognosis factors affecting OS and CSS,and a nomogram was constructed to predict OS and CSS.The clinical utility of columnar plots was assessed using calibration curves,area under the curve(AUC),and decision curve analysis(DCA).Result OS column line graphs were constructed based on nine independent predictors:age,tumor location,degree of differentiation,tumor size,TNM stage,chemotherapy,primary focus surgery,number of lymph nodes sampled,and serum carcinoembryonic antigen(CEA)level.The C-index of the nomogram to predict the 1-,3-,and 5-year OS were 0.764,0.790,and 0.805 in the training group,0.754,0.760,and 0.801 in the internal validation group,and 0.822,0.874,and 0.906 in the external validation group.CSS column line graphs were constructed based on 3 independent predictors of TNM staging,radiotherapy and chemotherapy.The 1-,3-,and 5-year CSS AUROC values of the training group were 0.791,0.757,and 0.782,respectively.0.682,0.709,0.625 in the internal validation group and 0.759,0.702,0.755 in the external validation group,respectively.The results of receiver operating characteristic curve(ROC),ROC and DCA showed that the use of our model was more effective in predicting OS and CSS than other single clinicopathological features.Conclusion In summary,the nomogram based on significant clinicopathological features can be conveniently used to predict OS and CSS individually in patients with CRCHBM.

Objective: To investigate the predictive value of D-dimer for deep venous thrombosis (DVT) of lower extremity in adult burn patients. Methods: A retrospective case series study was conducted. The clinical data of 3 861 adult burn patients who met the inclusion criteria and were admitted to the Department of Burns of Zhengzhou First People's Hospital from January 1, 2015 to December 31, 2019 were collected. The patients were divided into DVT group (n=77) and non-DVT group (n=3 784) according to whether DVT of lower extremity occurred during hospitalization or not. Data of patients in the two groups were collected and compared, including the gender, age, total burn area, D-dimer level, with lower limb burn and inhalation injury or not on admission, with sepsis/septic shock, femoral vein indwelling central venous catheter (CVC), history of surgery, and infusion of concentrated red blood cells or not during hospitalization. Data were statistically analyzed with independent sample t test, Mann-Whitney U test, and chi-square test. The indicators with statistically significant differences between the two groups were analyzed with multivariate logistic regression analysis to screen the independent risk factors for DVT of lower extremity in 3 861 adult burn patients. The receiver operating characteristic (ROC) curve of the independent risk factors predicting DVT of lower extremity in 3 861 adult burn patients were drawn, and the area under the curve (AUC), the optimal threshold value, and the sensitivity and specificity under the optimal threshold value were calculated. The quality of the AUC was compared by Delong test, and the sensitivity and specificity under the optimal threshold value were compared using chi-square test. Results: There were no statistically significant differences in gender, occurrence of sepsis/septic shock or history of surgery during hospitalization between patients in the two groups (P>0.05), while there were statistically significant differences in age, total burn area, D-dimer level, lower limb burn and inhalation injury on admission, and femoral vein indwelling CVC and infusion of concentrated red blood cells during hospitalization between patients in the two groups (t=-8.17, with Z values of -5.04 and -10.83, respectively, χ² values of 21.83, 5.37, 7.75, and 4.52, respectively, P<0.05 or P<0.01). Multivariate logistic regression analysis showed that age, total burn area, and D-dimer level were the independent risk factors for DVT of lower extremity in 3 861 adult burn patients (with odds ratios of 1.05, 1.02, and 1.14, respectively, 95% confidence intervals of 1.04-1.06, 1.00-1.03, and 1.10-1.20, respectively, P<0.05 or P<0.01). The AUCs of ROC of age, total burn area, and D-dimer level for predicting DVT of lower extremity in 3 861 adult burn patients were 0.74, 0.67, and 0.86, respectively (with 95% confidence intervals of 0.68-0.80, 0.60-0.74, and 0.83-0.89, respectively, P values<0.01), the optimal threshold values were 50.5 years old, 10.5% total body surface area, and 1.845 mg/L, respectively, the sensitivity under the optimal threshold values were 71.4%, 70.1%, and 87.0%, respectively, and the specificity under the optimal threshold values were 66.8%, 67.2%, and 72.9%, respectively. The AUC quality and sensitivity and specificity under the optimal threshold value of D-dimer level were significantly better than those of age (z=3.29, with χ² values of 284.91 and 34.25, respectively, P<0.01) and total burn area (z=4.98, with χ² values of 326.79 and 29.88, respectively, P<0.01), while the AUC quality and sensitivity and specificity under the optimal threshold values were similar between age and total burn area (P>0.05). Conclusions: D-dimer level is an independent risk factor for DVT of lower extremity in adult burn patients, its AUC quality and sensitivity and specificity under the optimal threshold value are better than those of age and total burn area, and it has good predictive value for DVT of lower extremity in adult burn patients.
Adult ; Burns/complications* ; Fibrin Fibrinogen Degradation Products/analysis* ; Humans ; Lower Extremity/blood supply* ; Lung Injury/etiology* ; Middle Aged ; Prognosis ; Retrospective Studies ; Shock, Septic/etiology* ; Venous Thrombosis/etiology*

Harmaline and harmine are β-carboline alkaloids with effective pharmacological effects. Harmaline can be transformed into harmine after oral administration. However, enzymes involved in the metabolic pathway remain unclear. In this study, harmaline was incubated with rat liver microsomes (RLM), rat brain microsomes (RBM), blood, plasma, broken blood cells, and heme peroxidases including horseradish peroxidase (HRP), lactoperoxidase (LPO), and myeloperoxidase (MPO). The production of harmine was determined by a validated UPLC-ESI-MS/MS method. Results showed that heme peroxidases catalyzed the oxidative dehydrogenation of harmaline. All the reactions were in accordance with the Hill equation. The reaction was inhibited by ascorbic acid and excess H₂O₂. The transformation of harmaline to harmine was confirmed after incubation with blood, plasma, and broken blood cells, rather than RLM and RBM. Harmaline was incubated with blood, plasma, and broken cells liquid for 3 h, and the formation of harmine became stable. Results indicated an integrated metabolic pathway of harmaline, which will lay foundation for the oxidation reaction of dihydro-β-carboline. Moreover, the metabolic stability of harmaline in blood should not be ignored when the pharmacokinetics study of harmaline is carried out.
Animals ; Harmaline/metabolism* ; Harmine/metabolism* ; Heme ; Hydrogen Peroxide ; Rats ; Tandem Mass Spectrometry

Based on the clinical needs and examination requirements of standardized training students, the China national standardized training textbook Acupuncture and Moxibustion for residents of traditional Chinese medicine has made innovations in the textbook content and form. In the part of meridians and acupoints, the classic original text is introduced and the main indications and operation methods of 200 commonly-used acupoints are summarized in the form of tables. In the part of acupuncture and moxibustion technique, the operating procedures are standardized and the core technical points of 20 commonly-used acupuncture and moxibustion techniques are summarized in the form of flow chart. In the part of acupuncture and moxibustion treatment, 48 typical diseases are introduced in the form of case discussion, highlighting the problem orientation and demonstrating the diagnosis and treatment procedures.
Acupuncture/education* ; Acupuncture Therapy ; China ; Humans ; Medicine, Chinese Traditional ; Moxibustion

OBJECTIVE: To investigate the clinicopathological features and prognosis of hormone receptor-positive (HR+), human epidermal growth factor receptor 2-negative (HER2 -) breast cancer. METHODS: In the study, 3 035 consecutive breast cancer patients diagnosed in Breast Disease Center, Peking University First Hospital from January 2008 to December 2017 were collected. The prognostic signi-ficance of important pathological factors in HR +/HER2 - patients with complete clinicopathological information was analyzed. RESULTS: Within the 1 920 (63.26%) cases of HR +/HER2 - breast cancer, there were 1 624 cases with complete clinicopathological data, of which, 124 cases (7.64%) recurred and/or metastasized and 63 cases died of the disease, and the 5-year overall survival (OS) rate and disease-free survival (DFS) rate was 93.0% and 92.6% respectively. The stage of pT1-2 was 92.80%, while pN0 was 69.03%. 89.66% cases belonged to histologically non-specific type and 30.11%, 55.60%, 14.29% were credited to Grade 1, 2 and 3 respectively. The distribution of ER negative, low or high expression groups were 1.60%, 2.09% and 96.31%, while PR were 6.83%, 10.47%, 82.70%, respectively. The group of Ki67 index < 10% was 19.52%, ≥10% & < 20% for 32.02%, ≥20% for 48.46%. Survival analysis showed that cases with pT1 stage had lower risk of recurrence than those with pT3, while cases with pT2 and pT3 had shorter DFS than those with pT1, with higher risk of recurrence and metastasis. Analysis proved that both pN stage and histological grade were negatively correlated with DFS. The cases with pN0, pN1 and pN2 were lower risk of recurrence than those with pN3, while cases with Grade 1 and 2 had lower risk of recurrence than cases with Grade 3. And the group of Ki67 index ≥20% showed higher risk of recurrence and metastasis. The prognostic significance of ER expression in HR+/HER2- breast cancer was not significant. However, the negative/low PR expression groups showed higher risk of recurrence and metastasis, of which PR < 10% group had shortest DFS and OS, followed by 10%-60% group and then > 60% group. The most common site of metastasis was bone (55 cases, 44.35%), while cases with liver metastasis (30 cases, 24.20%) had the worst outcome. CONCLUSION Our study revealed that pT, pN, Grade, HR expression level and Ki67 index were important prognostic factors for HR +/HER2 - breast cancer, although there are variables in prognostic value. Factors of pN and Grade showed independent prognostic significance. PR expression level had prognostic significance for the risk of recurrence and metastasis. The stratified level of PR expression (< 10%, 10%-60%, >60%) had independent prognostic value, showing successively longer DFS and OS, lower risk of recurrence. PR>60% group had the longest DFS and OS as well as the lowest risk of recurrence.
Breast Neoplasms ; Disease-Free Survival ; Female ; Humans ; Ki-67 Antigen/metabolism* ; Prognosis ; Receptor, ErbB-2/metabolism* ; Receptors, Progesterone/analysis* ; Triple Negative Breast Neoplasms/metabolism*

BACKGROUND: Methylene blue is the most commonly used tracer for sentinel lymph node (SLN) biopsy (SLNB) in China. This study aimed to investigate the feasibility of clinical application of SLNB using methylene blue dye (MBD) for early breast cancer and the prognosis of patients with different SLN and non-SLN statuses. METHODS: We retrospectively analyzed the clinicopathological data of patients with early breast cancer treated at the Peking University First Hospital between 2013 and 2018. We calculated the SLN identification rate (IR) in SLNB with MBD and the false-negative rate (FNR), and analyzed the prognosis of patients with different SLN and non-SLN statuses using Kaplan-Meier curves. RESULTS: Between January 2013 and December 2018, 1603 patients with early breast cancer underwent SLNB with MBD. The SLN IR was 95.8% (1536/1603). Two SLNs (median) were detected per patient. There were significant differences in FNR between patients with SLN micrometastasis and macrometastasis (19.0% vs. 4.5%, χ2 = 12.771, P < 0.001). Chi-square test showed that there were significant differences in SLN successful detection rates among patients with different vascular tumor embolism status (96.3% vs. 90.8%, χ2 = 9.013, P = 0.003) and tumor (T) stages (96.6% vs. 94.1%, χ2 = 5.189, P = 0.023). Multivariate analysis showed that vascular tumor embolism was the only independent factor for SLN successful detection (odds ratio: 0.440, 95% confidence interval: 0.224-0.862, P = 0.017). Survival analysis showed a significant difference in disease-free survival (DFS) between patients with non-SLN metastasis and patients without non-SLN metastasis (P = 0.006). CONCLUSION Our single-center data show that, as a commonly used tracer in SLNB in China, MBD has an acceptable SLN IR and a low FNR in frozen sections. This finding is consistent with reports of dual tracer-guided SLNB. Positive SLNs with non-SLN metastasis are associated with DFS.
Breast Neoplasms/surgery* ; China ; Humans ; Lymph Nodes ; Methylene Blue ; Retrospective Studies ; Sentinel Lymph Node Biopsy

Objective:To evaluate arthrography-assisted reduction in minimally invasive treatment of tibial plateau fractures.Methods:From January to May 2019, 9 patients with tibial plateau fracture were treated by arthrography-assisted minimally invasive surgery at Department of Orthopaedics and Traumatology, Hong-Hui Hospital. They were 6 males and 3 females with an average age of 45.5 years (from 27 to 63 years). According to the Schatzker classification, there were 6 cases of type Ⅱ, one of type Ⅲ and 2 cases of type Ⅳ. Arthrography was used to determine the lowest filling point of contrast medium after intra-articular injection and to ensure satisfactory reduction of articular surface after fracture reduction and bone grafting, followed by minimally invasive percutaneous plate implantation. Comparisons were made between preoperation and 6 months postoperation in the range of knee motion and knee scores of The Hospital for Special Surgery (HSS). Knee X-ray films were evaluated by the Rasmussen imaging system at 6 months postoperation. Collapse of articular surface was recorded pre- and post-operation as well as postoperative complications.Results:All the 9 patients were followed up for 6 to 9 months (average, 7 months). All patients achieved bony union within 12 weeks postoperation. The preoperative knee flexion (60.7°±13.1°) and HSS (51.9±5.7) were significantly improved to 122.4°±10.8° and 84.9±5.3 at 6 months postoperation ( P<0.05), but there was no significant difference between preoperation and 6 months postoperation in knee extension (4.4°±2.5° versus 4.6°±2.9°) ( P>0.05). The collapse of articular surface was improved from preoperative 9.5 mm (from 5 to 15 mm) to postoperative 1.3 mm (from 0 to 3 mm). The Rasmussen imaging scores at 6 months postoperation showed 6 excellent and 3 good. No such postoperative complications occurred as incision infection, joint stiffness, loosening or breakage of implants. Conclusion:Arthrography helps minimally invasive treatment of tibial plateau fracture because it can better display fracture collapse during operation and accurately judge indirect reduction of articular surface.

Objective:To establish a comet test method for detection of genotoxicity of three reference chemicals in rat liver cells. Methods:6-10 week old Sprague Dawley rats were randomly divided into 4 groups， with normal saline （0.9% NaCl solution） as negative control group. Animals in three test groups were treated， respectively， with ethyl methanesulfonate （EMS） 200 mg/kg， N-methyl-N-nitrosourea （MNU） 50 mg/kg， and D-mannitol 2 000 mg/kg. There were 10 animals in each group， 5 males and 5 females. The animals received two times （21 h interval） of test compounds through intragastric administration， and their clinical symptoms and body weight changes were recorded during the experiment. The rats were sacrificed 3 h after the last exposure. The liver was weighed， then used to prepare single-cell suspensions for the alkaline comet test which determines the average tail DNA content percentage （DNA%） of hepatocytes and other comet indicators. Results:（1） D-mannitol， EMS and MNU did not show significant toxicity in the whole animal. （2） The mean values of tail DNA content percentage （DNA%） of rat hepatocytes in EMS ［（60.07±24.69）%］ and MNU ［（41.66±22.35）%］ groups were higher than that in the negative control group ［（2.32±1.39）%］ and the difference was statistically significant （P<0.05）. The difference between D-mannitol group ［（3.06±3.30）%］ and the negative control group was not significant （P>0.05）. Conclusion:This laboratory has established a comet test method using hepatocytes from treated rats. Among three testing chemicals， EMS and MNU have displayed genotoxicity by this assay， but no genotoxicity was observed in D-mannitol treated animals.