ObjectiveTo investigate the effects of Scutellariae Radix combined with epidermal growth factor receptor-tyrosine kinase inhibitors （EGFR-TKIs） on cell proliferation， apoptosis， cancer stem cell （CSC） marker expression， and metabolism in non-small cell lung cancer （NSCLC） cells. MethodsThe anti-tumor effects of Scutellariae Radix and EGFR-TKIs （gefitinib or osimertinib） in NSCLC cells were evaluated using the cell counting kit-8 （CCK-8） and Annexin V-FITC/propidium iodide （PI） double staining apoptosis assay. The activity of Scutellariae Radix and EGFR-TKIs in three-dimensional （3D） cultures of NSCLC cells was assessed using the CellTiter-Glo® 3D cell viability assay. The mRNA and protein expression levels of CSC markers， sex determining region y box protein 2 （SOX2） and aldehyde dehydrogenase 1 family member A1 （ALDH1A1）， were detected by quantitative real-time polymerase chain reaction （Real-time PCR） and Western blot， respectively. Changes in intracellular reactive oxygen species （ROS） levels were detected by ROS staining， and the redox ratio was detected by femtosecond laser labeling free imaging （FLI）. ResultsUnder both two-dimensional （2D） and 3D culture conditions， compared with the blank group and EGFR-TKI group， the combination group showed significantly reduced cell viability and increased apoptosis rate （P<0.05）. Compared with the EGFR-TKI group， the mRNA and protein levels of CSC markers were significantly downregulated in the combination group （P<0.05）. Additionally， the redox ratio was significantly elevated （P<0.05）， and ROS levels were also increased in the combination group compared with the EGFR-TKI group. ConclusionIn NSCLC cells， Scutellariae Radix enhances the redox ratio and increases ROS levels， thereby inhibiting the expression of CSC markers and strengthening the anti-tumor effects of EGFR-TKIs. This provides a novel molecular mechanism by which Scutellariae Radix may enhance the sensitivity of targeted therapies.

OBJECTIVE: To investigate the hepatotoxicity of alkaloids from Euodiae Fructus combined with berberine (BBR) in Zuojin Pill, and to preliminarily explore the possible detoxification mechanism of the combination components. METHODS: The combination ratio of components was determined by the maximum concentration (Cmax) of the chemical components in Zuojin Pill. HepG2 cell model was used to investigate the combined toxicity of the hepatotoxic components from Euodiae Fructus, such as evodiamine (EVO) or dehydroevodiamine (DHED), with BBR for 48 h. The experimental groups were set as follows: the vehicle control group, the EVO group, the DHED group, the BBR group, and the combination group of EVO or DHED with BBR. The cell counting kit-8 (CCK-8) method was used to determine the cell viability, and the combination index (CI) was used to determine the combined toxicity of the components. The alanine transaminase (ALT), aspartate aminotransferase (AST), lactate dehydroge-nase (LDH), and alkaline phosphatase (ALP) activities as well as total bilirubin (TBIL) content in the cell culture supernatant were detected. The protein expression levels of bile acid transporters, such as bile salt export pump (BSEP) and multidrug resistance-associated protein 2 (MRP2), were detected by Western blot. The intracellular malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in HepG2 cells were detected. RESULTS: Compared with EVO or DHED group, the combination of EVO 1 μmol/L with BBR 10 μmol/L or DHED 50 μmol/L with BBR 35 μmol/L significantly increased cell viability of HepG2 cells (P < 0.01), with CI values of 77.89 or 4.49, respectively, much greater than 1. Significant decreases in the activities of ALT, AST, LDH, ALP, and TBIL content in the cell culture supernatant were found in both combination groups (P < 0.05, P < 0.01). Compared with the EVO group, the combination of EVO with BBR upregulated the protein expression levels of BSEP and MRP2. Compared with the DHED group, the combination of DHED with BBR significantly downregulated the protein expression levels of BSEP and MRP2 (P < 0.01). Compared with EVO or DHED group, the combination of EVO or DHED with BBR significantly reduced the MDA content in HepG2 cells (P < 0.05, P < 0.01). CONCLUSION A certain ratio of BBR combined with EVO or DHED had an antagonistic effect on HepG2 cytotoxicity, which might be related to regulating the expression of bile acid transpor-ters, and reducing lipid peroxidation damage.
Humans ; Hep G2 Cells ; Berberine/pharmacology* ; Drugs, Chinese Herbal/toxicity* ; Evodia/chemistry* ; Alkaloids/pharmacology* ; Cell Survival/drug effects* ; Multidrug Resistance-Associated Proteins/metabolism* ; Multidrug Resistance-Associated Protein 2 ; Quinazolines

The biological clock(also known as the circadian rhythm)is the fundamental reliance for all organisms on Earth to adapt and survive in the Earth's rotation environment.Circadian rhythm is the most basic regulatory mechanism of life activities,and plays a key role in maintaining normal physiological and biochemical homeostasis,disease occurrence and treatment.Recent studies have shown that the biologi-cal clock plays an important role in the development of oral tissues and in the occurrence and treatment of oral diseases.Since there is cur-rently no guiding literature on the research methods of biological clock in stomatology,researchers mainly conduct research based on pub-lished references,which has led to controversy about the research methods of biological clock in stomatology,and there are many confusions about how to rationally apply the research methods of circadia rhythms.In view of this,this expert consensus summarizes the characteristics of the biological clock and analyzes the shortcomings of the current biological clock research in stomatology,and organizes relevant experts to summarize and recommend 10 principles as a reference for the rational implementation of the biological clock in stomatology research.

Purpose: This study aimed to compare and analyze the feasibility and long-term efficacy of prostatic capsule-sparing (PCS) and nerve-sparing (NS) radical cystectomy in the treatment of bladder cancer. Methods: From June 2004 to December 2021, our institution treated and followed 145 patients who underwent radical cystectomy with neobladder reconstruction for over a year. These patients were divided into 2 groups: PCS (n=74) and NS (n=71). To minimize potential biases, 1:1 propensity score matching was utilized to compare oncological outcomes, functional outcomes, and complications between the groups. Additionally, Kaplan-Meier analysis and the log-rank test were used to evaluate survival differences between the PCS and NS groups. Results: The median follow-up durations for PCS and NS were 155 and 122 months, respectively. After adjusting for propensity scores, a total of 96 patients (48 in each group) were included for further analysis. Kaplan-Meier curves showed no statistically significant differences in metastasis-free probability (P=0.206), cancer-specific survival (P=0.091), and overall survival (P=0.208). The daytime urinary control (UC) rate at 3, 6, and 12 months postoperatively was 72.9%, 91.7%, and 97.9% in the PCS group and 47.9%, 79.2%, and 91.7% in the NS group, respectively (P=0.012, P=0.083, and P=0.362). The nocturnal UC rate was 54.2%, 85.4%, and 95.8% in the PCS group, and 31.3%, 60.4%, and 83.3% in the NS group, respectively (P=0.023, P=0.006, and P=0.091). Regarding erectile function recovery, 62.5% of patients in the PCS group and 22.9% in the NS group returned to preoperative levels (P<0.001). Conclusions PCS outperformed NS in restoring UC and sexual function and did not affect oncological outcomes. However, PCS was associated with a higher risk of complications linked to bladder-neck obstruction.

Purpose: This study aimed to compare and analyze the feasibility and long-term efficacy of prostatic capsule-sparing (PCS) and nerve-sparing (NS) radical cystectomy in the treatment of bladder cancer. Methods: From June 2004 to December 2021, our institution treated and followed 145 patients who underwent radical cystectomy with neobladder reconstruction for over a year. These patients were divided into 2 groups: PCS (n=74) and NS (n=71). To minimize potential biases, 1:1 propensity score matching was utilized to compare oncological outcomes, functional outcomes, and complications between the groups. Additionally, Kaplan-Meier analysis and the log-rank test were used to evaluate survival differences between the PCS and NS groups. Results: The median follow-up durations for PCS and NS were 155 and 122 months, respectively. After adjusting for propensity scores, a total of 96 patients (48 in each group) were included for further analysis. Kaplan-Meier curves showed no statistically significant differences in metastasis-free probability (P=0.206), cancer-specific survival (P=0.091), and overall survival (P=0.208). The daytime urinary control (UC) rate at 3, 6, and 12 months postoperatively was 72.9%, 91.7%, and 97.9% in the PCS group and 47.9%, 79.2%, and 91.7% in the NS group, respectively (P=0.012, P=0.083, and P=0.362). The nocturnal UC rate was 54.2%, 85.4%, and 95.8% in the PCS group, and 31.3%, 60.4%, and 83.3% in the NS group, respectively (P=0.023, P=0.006, and P=0.091). Regarding erectile function recovery, 62.5% of patients in the PCS group and 22.9% in the NS group returned to preoperative levels (P<0.001). Conclusions PCS outperformed NS in restoring UC and sexual function and did not affect oncological outcomes. However, PCS was associated with a higher risk of complications linked to bladder-neck obstruction.

Purpose: This study aimed to compare and analyze the feasibility and long-term efficacy of prostatic capsule-sparing (PCS) and nerve-sparing (NS) radical cystectomy in the treatment of bladder cancer. Methods: From June 2004 to December 2021, our institution treated and followed 145 patients who underwent radical cystectomy with neobladder reconstruction for over a year. These patients were divided into 2 groups: PCS (n=74) and NS (n=71). To minimize potential biases, 1:1 propensity score matching was utilized to compare oncological outcomes, functional outcomes, and complications between the groups. Additionally, Kaplan-Meier analysis and the log-rank test were used to evaluate survival differences between the PCS and NS groups. Results: The median follow-up durations for PCS and NS were 155 and 122 months, respectively. After adjusting for propensity scores, a total of 96 patients (48 in each group) were included for further analysis. Kaplan-Meier curves showed no statistically significant differences in metastasis-free probability (P=0.206), cancer-specific survival (P=0.091), and overall survival (P=0.208). The daytime urinary control (UC) rate at 3, 6, and 12 months postoperatively was 72.9%, 91.7%, and 97.9% in the PCS group and 47.9%, 79.2%, and 91.7% in the NS group, respectively (P=0.012, P=0.083, and P=0.362). The nocturnal UC rate was 54.2%, 85.4%, and 95.8% in the PCS group, and 31.3%, 60.4%, and 83.3% in the NS group, respectively (P=0.023, P=0.006, and P=0.091). Regarding erectile function recovery, 62.5% of patients in the PCS group and 22.9% in the NS group returned to preoperative levels (P<0.001). Conclusions PCS outperformed NS in restoring UC and sexual function and did not affect oncological outcomes. However, PCS was associated with a higher risk of complications linked to bladder-neck obstruction.

Objective:To analyze and predict the transmission risk of visceral leishmaniasis in Gansu Province based on an ecological niche model, providing a basis for the development of precise prevention and control measures and epidemic surveillance.Methods:The information of reported cases of visceral leishmaniasis in Gansu Province from 2015 to 2021 were collected from the National Infectious Disease Reporting Information Management System, and the longitude and latitude coordinates of the distribution points of cases and the data of 19 climate variables, 5 geographical variables and 2 socio-economic variables within the region were obtained. Based on an ecological niche model, a model for predicting the transmission risk of visceral leishmaniasis was constructed using the maximum entropy algorithm (MaxEnt), and its performance was evaluated by the area under the receiver operating characteristic curve (AUC). Then the importance of each environmental variable of the model was evaluated, and the distribution area of visceral leishmaniasis transmission risk in Gansu Province was predicted.Results:A total of 368 cases of visceral leishmaniasis were reported in Gansu Province from 2015 to 2021, of which 89.13% (328/368) were from Longnan City and Gannan Tibetan Autonomous Prefecture (Gannan Prefecture). The number of cases peaked in 2017 (79 cases, 21.47%). The model had high prediction accuracy (AUC = 0.985). The results of model analysis showed that the important climate variable affecting the distribution of visceral leishmaniasis was the average temperature in the coldest quarter (contribution value of 3.1), the geographical variables were land use type (contribution value of 52.6) and vegetation cover type (contribution value of 8.5), and the socio-economic variable was population size (contribution value of 14.3). The distribution results of transmission risk showed that high, medium and low risk areas exhibited a gradual transition from the southern part to the northwest part of Gansu Province. The high risk areas were mainly located in the central and southern parts of Longnan City and the southern part of Gannan Prefecture, accounting for 0.18% of the total area of the province. Medium and low risk areas accounted for 0.48% and 2.47% of the total area of the province, respectively; and areas with no risk accounted for 96.87%.Conclusions:The ecological niche model predicts that the spread of visceral leishmaniasis in Gansu Province is characterized by point like dispersion and local high aggregation distribution. It is necessary to strengthen monitoring and prevention and control of high-risk areas such as Longnan City and Gannan Prefecture.

This article reported a male neonate with lethal mitochondrial trifunctional protein deficiency (MTPD) caused by compound heterozygous variations in the HADHB gene. The patient presented with poor milk intake complicated by abnormal myocardial enzymes within 24 h after birth and was transferred to the Children's Hospital of Nanjing Medical University on day 4. Physical examination revealed no obvious abnormalities on admission. Laboratory examination showed increased creatine kinase isoenzyme and cardiac troponin levels, and electrocardiogram suggested sinus tachycardia and low QRS voltage in limb leads. Blood screening for metabolic abnormalities showed high levels of tetradecenyl carnitine and various 3-hydroxycarnitines. Heterozygous mutations of c.739C>T(p.Arg247Cys) and c.607C>T(p.Arg203Ter,272) were detected in the HADHB gene in the boy, which were pathogenic variants included in the Human Gene Mutation Database. Followed up to three months of age, the boy was readmitted to hospital due to poor milk intake for one week and poor response for 2 d after catching a cold. After admission, he quickly developed multiple organs dysfunction such as heart failure and respiratory failure, and then died. Lethal MTPD is rare with no effective treatment and poor prognosis. Lethal MTPD should be highly suspected when unexplained cardiomyopathy, hypoglycemia, acidosis and other metabolic abnormalities appear in the neonatal period, and an early diagnosis could be confirmed with genetic testing in the neonatal period.

Objective:To investigate the preventive and therapeutic effects of resveratrol on lens opacification in diabetic rats and its biological mechanism.Methods:Fifty 8-week-old healthy male SPF grade SD rats were selected and randomly divided into blank control group, model group, gliclazide group, low-dose resveratrol group and high-dose resveratrol group according to their body weight, with 10 rats in each group.The diabetes model was established by intraperitoneal injection of streptozotocin in model group, gliclazide group, low-dose resveratrol group and high-dose resveratrol group.On the third day after modeling, rats in gliclazide group was gavaged with 2 mg/(kg·d) gliclazide suspension, and rats in low-dose and high-dose resveratrol groups were gavaged with 20 and 40 mg/(kg·d) resveratrol, respectively, for four weeks.Rats in blank control group and model group were gavaged with the same volume of normal saline once a day, also for four weeks.After the diabetes model was established, there were 10 rats in blank control group and 9 rats in the other four groups.The fasting blood glucose concentration of the rats was measured with a blood glucose meter.The concentrations of fasting insulin, superoxide dismutase (SOD) 1, SOD2, SOD3, and glutathione peroxidase (GPX1) were determined by enzyme-linked immunosorbent assay.Lens opacification after treatment was observed by slit lamp microscopy.Morphologic changes in lens cells were examined by hematoxylin-eosin staining.Apoptosis of lens epithelial cells (LECs) was detected using TUNEL.The relative expressions of nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins in lens tissues were determined by Western blot.The study protocol was approved by the Welfare Ethics Committee of Experimental Animal of Zhengzhou University (No.IACYC2019-02).Results:Fasting blood glucose concentration, fasting insulin level, and apoptosis rate of LECs were increased and the concentrations of SOD1, SOD2, SOD3, and GPX1 were decreased in model group in comparison with blank control group, and the differences were statistically significant (all at P<0.05). Fasting blood glucose concentration, fasting insulin level, and apoptosis rate of LECs were decreased and the concentrations of SOD1, SOD2, SOD3, and GPX1 were increased in gliclazide group, low-dose resveratrol group, and high-dose resveratrol group compared with model group, and the differences were statistically significant (all at P<0.05). Fasting blood glucose concentration, fasting insulin level, and apoptosis rate of LECs were decreased and the concentrations of SOD1, SOD2, SOD3, and GPX1 were increased in gliclazide group and high-dose resveratrol group compared with low-dose resveratrol group, and the differences were statistically significant (all at P<0.05). The proportions of grade 0, 1 and 2 lens opacities after treatment were 100.00%, 0.00% and 0.00% in blank control group, 0.00%, 66.67% and 33.33% in model group, 77.78%, 22.22% and 0.00% in gliclazide group, 22.22%, 44.44% and 33.33% in low-dose resveratrol group, and 66.67%, 33.33% and 0.00% in high-dose resveratrol group, respectively, with a statistically significant difference ( H=7.514, P<0.001). Compared with model group, lens opacification was less severe in blank control group, gliclazide group, low-dose resveratrol group, and high-dose resveratrol group, with statistically significant differences (all at P<0.05). Lens opacification was less severe in gliclazide group and high-dose resveratrol group compared with low-dose resveratrol group, showing statistically significant differences (both at P<0.05). Compared with model group, there were fewer abnormal changes of lens cells and sub-organelles in gliclazide group, low-dose resveratrol group and high-dose resveratrol group, and the abnormalities in gliclazide group and high-dose resveratrol group were slighter.Compared with model group, the relative expression levels of Nrf2 and HO-1 were higher in blank control group, gliclazide group, low-dose resveratrol group, and high-dose resveratrol group, with statistically significant differences (all at P<0.05). The relative expression levels of Nrf2 and HO-1 were higher in gliclazide group and high-dose resveratrol group compared with low-dose resveratrol group, showing statistically significant differences (both at P<0.05). Conclusions:Resveratrol can reduce lens opacification in diabetic rats and its mechanism may be related to the regulation of the Nrf2/HO-1 signaling pathway by exerting antioxidative stress effects.

Objective:To study the main blood parameters of Leishmania-infected dogs with different clinical symptoms in the high incidence area of visceral leishmaniasis in Gansu Province, and to provide scientific basis for early diagnosis and monitoring of canine leishmaniasis. Methods:In May 2019, Wenxian County of Longnan City and Diebu County of Gannan Tibetan Autonomous Prefecture, the historical endemic areas of visceral leishmaniasis in Gansu Province, were selected as the investigation sites, and 40 dogs with positive Leishmania DNA were investigated. According to their clinical symptoms, they were divided into three groups: asymptomatic ( n = 17), minimally symptomatic ( n = 12) and symptomatic ( n = 11). At the same time, 10 healthy dogs were selected as controls. Major blood indexes were detected by an automatic animal blood five-classification cell analyzer. Results:Compared with the control group, the number of red blood cells, hemoglobin content, hematocrit, and the number of lymphocytes, monocytes and eosinophils in the asymptomatic, minimally symptomatic and symptomatic groups were decreased ( P < 0.05), while the number of white blood cells and neutrophils in minimally symptomatic and symptomatic groups were decreased ( P < 0.05). The above blood indexes were lower in the symptomatic group than those in the asymptomatic group. The number of red blood cells (control group vs symptomatic group) decreased from (6.82 ± 0.05) × 10 12/L to (4.19 ± 0.08) × 10 12/L, hemoglobin content decreased from (15.84 ± 0.74) g/L to (9.65 ± 0.18) g/L, hematocrit decreased from (46.41 ± 0.95)% to (27.86 ± 0.42)%, the number of white blood cells decreased from (14.90 ± 0.22) × 10 9/L to (10.23 ± 0.24) × 10 9/L, the number of neutrophils decreased from (7.25 ± 0.18) × 10 9/L to (6.18 ± 0.11) × 10 9/L, the number of lymphocytes decreased from (2.80 ± 0.07) × 10 9/L to (1.64 ± 0.06) × 10 9/L, the number of monocytes decreased from (1.13 ± 0.01) × 10 9/L to (0.53 ± 0.02) × 10 9/L, and the number of eosinophils decreased from (1.70 ± 0.06) × 10 9/L to (0.71 ± 0.03) × 10 9/L. Conclusion:Different clinical symptoms of Leishmania-infected dogs have different blood test results.