1.Raman Spectroscopy Analysis of The Temporal Heterogeneity in Lung Cell Carcinogenesis Induced by Benzo(a)pyrene
Hai-Tao ZHOU ; Wei YAO ; Cao-Zhe CUI ; Xiao-Tong ZHOU ; Xi-Long LIANG ; Cheng-Bing QIN ; Lian-Tuan XIAO ; Zhi-Fang WU ; Si-Jin LI
Progress in Biochemistry and Biophysics 2024;51(6):1458-1470
ObjectiveTemporal heterogeneity in lung cancer presents as fluctuations in the biological characteristics, genomic mutations, proliferation rates, and chemotherapeutic responses of tumor cells over time, posing a significant barrier to effective treatment. The complexity of this temporal variance, coupled with the spatial diversity of lung cancer, presents formidable challenges for research. This article will pave the way for new avenues in lung cancer research, aiding in a deeper understanding of the temporal heterogeneity of lung cancer, thereby enhancing the cure rate for lung cancer. MethodsRaman spectroscopy emerges as a powerful tool for real-time surveillance of biomolecular composition changes in lung cancer at the cellular scale, thus shedding light on the disease’s temporal heterogeneity. In our investigation, we harnessed Raman spectroscopic microscopy alongside multivariate statistical analysis to scrutinize the biomolecular alterations in human lung epithelial cells across various timeframes after benzo(a)pyrene exposure. ResultsOur findings indicated a temporal reduction in nucleic acids, lipids, proteins, and carotenoids, coinciding with a rise in glucose concentration. These patterns suggest that benzo(a)pyrene induces structural damage to the genetic material, accelerates lipid peroxidation, disrupts protein metabolism, curtails carotenoid production, and alters glucose metabolic pathways. Employing Raman spectroscopy enabled us to monitor the biomolecular dynamics within lung cancer cells in a real-time, non-invasive, and non-destructive manner, facilitating the elucidation of pivotal molecular features. ConclusionThis research enhances the comprehension of lung cancer progression and supports the development of personalized therapeutic approaches, which may improve the clinical outcomes for patients.
2.A national questionnaire survey on endoscopic treatment for gastroesophageal varices in portal hypertension in China
Xing WANG ; Bing HU ; Yiling LI ; Zhijie FENG ; Yanjing GAO ; Zhining FAN ; Feng JI ; Bingrong LIU ; Jinhai WANG ; Wenhui ZHANG ; Tong DANG ; Hong XU ; Derun KONG ; Lili YUAN ; Liangbi XU ; Shengjuan HU ; Liangzhi WEN ; Ping YAO ; Yunxiao LIANG ; Xiaodong ZHOU ; Huiling XIANG ; Xiaowei LIU ; Xiaoquan HUANG ; Yinglei MIAO ; Xiaoliang ZHU ; De'an TIAN ; Feihu BAI ; Jitao SONG ; Ligang CHEN ; Yingcai MA ; Yifei HUANG ; Bin WU ; Xiaolong QI
Chinese Journal of Digestive Endoscopy 2024;41(1):43-51
Objective:To investigate the current status of endoscopic treatment for gastroesophageal varices in portal hypertension in China, and to provide supporting data and reference for the development of endoscopic treatment.Methods:In this study, initiated by the Liver Health Consortium in China (CHESS), a questionnaire was designed and distributed online to investigate the basic condition of endoscopic treatment for gastroesophageal varices in portal hypertension in 2022 in China. Questions included annual number and indication of endoscopic procedures, adherence to guideline for preventing esophagogastric variceal bleeding (EGVB), management and timing of emergent EGVB, management of gastric and isolated varices, and improvement of endoscopic treatment. Proportions of hospitals concerning therapeutic choices to all participant hospitals were calculated. Guideline adherence between secondary and tertiary hospitals were compared by using Chi-square test.Results:A total of 836 hospitals from 31 provinces (anotomous regions and municipalities) participated in the survey. According to the survey, the control of acute EGVB (49.3%, 412/836) and the prevention of recurrent bleeding (38.3%, 320/836) were major indications of endoscopic treatment. For primary [non-selective β-blocker (NSBB) or endoscopic therapies] and secondary prophylaxis (NSBB and endoscopic therapies) of EGVB, adherence to domestic guideline was 72.5% (606/836) and 39.2% (328/836), respectively. There were significant differences in the adherence between secondary and tertiary hospitals in primary prophylaxis of EGVB [71.0% (495/697) VS 79.9% (111/139), χ2=4.11, P=0.033] and secondary prophylaxis of EGVB [41.6% (290/697) VS 27.3% (38/139), χ2=9.31, P=0.002]. A total of 78.2% (654/836) hospitals preferred endoscopic therapies treating acute EGVB, and endoscopic therapy was more likely to be the first choice for treating acute EGVB in tertiary hospitals (82.6%, 576/697) than secondary hospitals [56.1% (78/139), χ2=46.33, P<0.001]. The optimal timing was usually within 12 hours (48.5%, 317/654) and 12-24 hours (36.9%, 241/654) after the bleeding. Regarding the management of gastroesophageal varices type 2 and isolated gastric varices type 1, most hospitals used cyanoacrylate injection in combination with sclerotherapy [48.2% (403/836) and 29.9% (250/836), respectively], but substantial proportions of hospitals preferred clip-assisted therapies [12.4% (104/836) and 26.4% (221/836), respectively]. Improving the skills of endoscopic doctors (84.2%, 704/836), and enhancing the precision of pre-procedure evaluation and quality of multidisciplinary team (78.9%, 660/836) were considered urgent needs in the development of endoscopic treatment. Conclusion:A variety of endoscopic treatments for gastroesophageal varices in portal hypertension are implemented nationwide. Participant hospitals are active to perform emergent endoscopy for acute EGVB, but are inadequate in following recommendations regarding primary and secondary prophylaxis of EGVB. Moreover, the selection of endoscopic procedures for gastric varices differs greatly among hospitals.
3.Endothelin-1 regulates SOCC/TGF-β and involved in atrial fibrosis in rats with atrial fibrillation
Zhuoran JIA ; Manyu DAI ; Shichu LIANG ; Jian WU ; Yangcheng XUE ; Dingxin ZHANG ; Bing SHEN ; Ren ZHAO
Acta Universitatis Medicinalis Anhui 2024;59(3):429-435
Objective To investigate the effect and mechanism of endothelin-1(ET-1)on atrial fibrosis in Atrial fibrillation(AF)rats.Methods Fourteen adult male SD rats were randomly divided into normal control(NC)group and Atrial fibrillation(AF)group.The rat model of Atrial fibrillation was established by injecting 0.1 ml/100g CaCl2-Ach mixture into the tail vein once a day for one week.The control group was injected with the same dose of normal saline.An electrocardiogram of normal or atrial fibrillation was recorded on the first day and the eighth day in each group,and echocardiography was used to monitor atrial size and cardiac function.The fibrosis of atrial was observed using Masson and HE staining.The expression of endothelin-1(ET-1),collagen-I(Col-I),transforming growth factor-β(TGF-β)and the store operated calcium channel(SOCC)protein Orai1,stromal in-teraction molecule 1(STIM1)in atrial tissue were detected by Western blot.HL-1 cells were cultured and treated with gradient concentration of ET-1 for 24 hours.Western blot was used to observe changes in the expression of TGF-β,Orai1 and STIM1 proteins in ET-1/SOCC/TGF-β signaling pathway of HL-1 cells.Small interfering RNA(siRNA)transfection method was used to knock down the expression of Orai1 in HL-1 cells,then the cells were treated with appropriate concentrations of ET-1 for 24 hours,and the expression of TGF-β protein in HL-1 cells was detected by Western blot.Results Compared with the control group,echocardiography showed a significant in-crease in left atrial diameter(LAD)of the heart in atrial fibrillation rats(P<0.05).The HE and Masson staining results showed significant fibrosis in the myocardial tissue of AF group rats(P<0.05),and the Western blot re-sults indicated the expression of ET-1,Orai1,STIM1,TGF-β and COL-Ⅰ in the myocardial tissue of AF group significantly increased compared to the NC group(P<0.05).After ET-1 treatment of HL-1 cells,the protein ex-pression of Orai1,STIM1and TGF-β increased(P<0.05),while knocking down Orai1 in HL-1 cells,ET-1 treat-ment no longer caused the expression of TGF-β a significant upregulation.Conclusion AF caused by atrial fibril-lation results in a significant increase in ET-1 expression in atrial tissue,and ET-1/SOCC/TGF-β signal pathway promotes atrial fibrillation and fibrosis.
4.Analysis of the correlation between the clinical features of 1 865 children and adolescents with supernumerary teeth and the selection of anesthesia methods
ZHANG Yaqiu ; FENG Caihua ; LIANG Lirong ; LIU Fei ; WU Long ; WANG Peijuan ; GAO Zhenzhen ; LIU Bing
Journal of Prevention and Treatment for Stomatological Diseases 2024;32(12):954-962
Objective:
To retrospectively analyze the epidemiological characteristics of supernumerary teeth in patients aged 4-18 years old and the influencing factors on the selection of anesthesia methods, and to provide a reference for the selection of anesthesia plans for children and adolescents with supernumerary teeth.
Methods:
This study is a retrospective study approved by the Institutional Ethics Committee. Based on clinical electronic medical record system and cone beam CT (CBCT) data, a retrospective analysis was conducted on 2 210 patients 4-18 years of age who underwent supernumerary tooth extraction at the School of Stomatology, Fourth Military Medical University from August 2019 to July 2021. Inclusion criteria: ① Age 4-18 years old; and ② The American Society of Anesthesiologists (ASA) classifies anesthesia into grades I-II; and ③ Have complete oral and anesthesia case records and relevant imaging data. Exclusion criteria: ① Incomplete medical records or unclear imaging data; and ② Patients with ASA grade II or above. Patients’ gender and age, the number of supernumerary teeth, arch, location, orientation, eruption status, reason for appointment, anxiety level, degree of cooperation, anesthesia method, and other relevant information were collected and statistically analyzed.
Results:
A total of 1 865 eligible patients were included, with an average age of (8.9±3.2) years old. There were more male patients (71.37%, 1 331 cases) than female patients (28.63%, 534 cases) (P < 0.001), with a gender ratio of 2.49:1. The majority of supernumerary teeth were single (75.97%, 1 417/1 865), primarily located in the maxilla (97.2%, 1 812/1 865) and the anterior dental region (94.2%, 1 757/1 865), and in a centric position (77.3%, 1 442/1 865). The majority of patients with erupted supernumerary teeth were active in seeking treatment (97.67%, 335/343). Patients with supernumerary teeth located in the maxilla and mandible bones, as well as in the nasal floor, were mainly referred to the department for diagnosis, accounting for 94.38%(1 361/1 442) and 90.00% (72/80) (χ2=1 363.24, P < 0.001), respectively. Regarding anesthesia methods, the largest proportion of patients received nitrous oxide sedation-assisted local anesthesia or nerve block anesthesia, accounting for 38.07% (710/1 865), followed by local anesthesia, accounting for 35.23% (657/1 865). The proportion of patients receiving midazolam intravenous sedation with local anesthesia or nerve block anesthesia and general anesthesia was relatively small, accounting for 20.86% (389/1 865) and 5.84% (109/1 865). Patients 13-18 years of age with supernumerary teeth in the mandibular and posterior regions and without anxiety had the highest proportion of local anesthesia use (P < 0.001). In contrast, patients who had supernumerary teeth located at the base of the nose (50%), severe anxiety (94.12%), and poor cooperation (98.18%) had the highest proportion of general anesthesia use (P < 0.001). There was no significant difference (P = 0.35) in the incidence of intraoperative and postoperative complications after the extraction of supernumerary teeth. However, the proportion of anesthesia-related complications, such as dizziness, coughing, and respiratory depression, occurring in patients who received general anesthesia was higher than local anesthesia, accounting for 3.81% (P = 0.006).
Conclusion
There is a gender difference in the incidence of supernumerary teeth in patients 4-18 years of age, with a higher prevalence in males. The majority of supernumerary teeth are single and located in the maxillary anterior region, predominantly in a centric position. Patients whose teeth had erupted were more likely to seek medical treatment voluntarily, while patients with supernumerary teeth located in the maxilla and mandible bones, as well as in the nasal floor were more likely to be referred to the department. Patients with high levels of anxiety, poor cooperation, young age, multiple teeth, and high surgical difficulty were more inclined to choose general anesthesia.
5.Efficacy of posterior reduction and fixation combined with pedicle subtraction osteotomy in the treatment of ankylosing spondylitis kyphotic deformity complicated by upper cervical spine injury
Jian ZHANG ; Fangsheng HE ; Jun SHENG ; Rui CAO ; Tao XU ; Weidong LIANG ; Bing WU ; Weibin SHENG
Chinese Journal of Trauma 2024;40(6):522-530
Objective:To investigate the efficacy of posterior reduction and fixation combined with pedicle subtraction osteotomy (PSO) in the treatment of ankylosing spondylitis (AS) kyphotic deformity complicated by upper cervical spine injury.Methods:A retrospective case series study was conducted to analyze the clinical data of 8 patients with AS kyphotic deformity complicated by upper cervical spine injury, who were admitted to First Affiliated Hospital of Xinjiang Medical University from October 2010 to December 2022. All were males, aged 32-58 years [(46.9±8.7)years]. Acute injury was found in 1 patient and chronic injuries in 7 patients, including 3 with cervicothoracic kyphotic deformity and 5 with thoracolumbar kyphotic deformity. Five patients were complicated by odontoid fractures, 1 by C 2-C 3 fracture and 2 by atlantoaxial dislocation. According to American Spinal Injury Association (ASIA) scale, 2 patients were with grade C, 3 with grade D, and 3 with grade E. The patients were treated with posterior upper cervical reduction and fixation combined with cervicothoracic PSO or thoracolumbar PSO respectively according to the location of the deformity. The operation time and intraoperative blood loss were recorded. Neck disability index (NDI), visual analogue scale (VAS), C 0-C 2 angle, C 1-C 2 angle, cervical lordosis angle (CL), head tilt angle (HT), chin-brow vertical angle (CBVA), cervical sagittal vertical axis (CSVA), and sagittal vertical axis (SVA) were compared before surgery, at 1 week and 3, 6, 12 months after surgery, and at the last follow-up. The healing of fractures and fusion of osteotomy site were recorded at 12 months after surgery. Recovery of neurological function was observed at the last follow-up. Incidence of complications was observed. Results:Patients were all followed up for 12-24 months [(17.0±5.4)months]. The operation time was 5.5-7.2 hours [(6.2±0.6)hours] and the intraoperative blood loss was 480-800 ml [(629.4±124.0)ml]. The NDI scores at 1 week and 3, 6, 12 months after surgery, and at the last follow-up were (14.6±2.6)points, (13.6±2.8)points, (12.8±2.4)points, (12.8±2.7)points, and (12.8±2.6)points respectively, significantly lower than (29.6±8.5)points preoperatively ( P<0.01). There were no significant differences in NDI scores at various time points after surgery ( P>0.05). The VAS scores were 2.0(1.0, 3.0)points, 1.5(1.0, 2.0)points, 0.5(0.0, 1.8)points, 0.5(0.0, 1.7)points, and 0.5(0.0, 1.8)points respectively, significantly lower than 3.5(3.0, 4.8)points preoperatively ( P<0.01). The VAS score at 3 months postoperatively was lower than that at 1 week postoperatively ( P<0.05), and the VAS score at 6 months postoperatively was lower than that at 3 months postoperatively ( P<0.05). There were no statistically significant differences in VAS scores at 6, 12 months postoperatively, and at the last follow-up ( P>0.05). There were no statistically significant differences between C 0-C 2 angle and C 1-C 2 angle preoperatively and at different time points postoperatively ( P>0.05). At 1 week and 3, 6, 12 months postoperatively and at the last follow-up, the CL values were -8.5(-5.3, -11.9)°, -8.6(-5.5, -11.9)°, -8.4(5.2, -12.1)°, -8.8(-5.6, -12.4)°, and -8.7(-5.3, -12.5)° respectively, significantly higher than 1.2(9.5, -4.8)° preoperatively ( P<0.01); the HT values were 6.1(4.5, 9.6)°, 6.1(4.3, 9.4)°, 6.0(4.2, 8.9)°, 6.0(4.2, 9.2)°, and 6.1(4.3, 9.2)° respectively, significantly lower than 17.0(10.3, 22.0)° preoperatively ( P<0.01); the CBVA values were (23.2±5.0)°, (23.1±4.8)°, (23.0±4.7)°, (23.1±4.7)°, and (23.1±4.9)° respectively, significantly lower than (44.1±9.8)° preoperatively ( P<0.01); the CSVA values were 5.2(4.2, 7.5)cm, 5.4(4.1, 7.1)cm, 4.7(4.0, 7.4)cm, 5.4(4.1, 7.0)cm, and 5.1(4.3, 6.5)cm respectively, significantly shorter than 9.0(7.8, 9.3)cm preoperatively ( P<0.01); the SVA values were 7.7(6.2, 13.7)cm, 7.5(6.0, 13.4)cm, 7.6(6.2, 13.2)cm, 7.4(6.3, 13.1)cm, and 7.5(6.2, 13.2)cm respectively, significantly shorter than 16.8(8.2, 27.2)cm preoperatively ( P<0.05). There were no statistically significant differences among CL, HT, CBVA, CSVA, and SVA values at different time points after surgery ( P>0.05). All the fractures healed at 12 months after surgery and Bridwell grade I healing was achieved at all the osteotomy sites. At the last follow-up, all the patients were classified as ASIA grade E, significantly improved compared with preoperatively ( P<0.01). One patient had transient C 8 nerve paralysis after surgery and recovered after 4 weeks′ treatment. The remaining patients did not develop any infection or internal fixation-related complications, such as broken rods, broken screws, or loose screws. Conclusion:For AS kyphosis deformity complicated with upper cervical injury, posterior reduction and fixation combined with cervicothoracic or thoracolumbar PSO can effectively promote functional recovery, relieve pain, maintain overall trunk balance, improve neurological symptoms, and reduce the incidence of complications.
6.Risk factors and predictive model of cerebral edema after road traffic accidents-related traumatic brain injury
Di-You CHEN ; Peng-Fei WU ; Xi-Yan ZHU ; Wen-Bing ZHAO ; Shi-Feng SHAO ; Jing-Ru XIE ; Dan-Feng YUAN ; Liang ZHANG ; Kui LI ; Shu-Nan WANG ; Hui ZHAO
Chinese Journal of Traumatology 2024;27(3):153-162
Purpose::Cerebral edema (CE) is the main secondary injury following traumatic brain injury (TBI) caused by road traffic accidents (RTAs). It is challenging to be predicted timely. In this study, we aimed to develop a prediction model for CE by identifying its risk factors and comparing the timing of edema occurrence in TBI patients with varying levels of injuries.Methods::This case-control study included 218 patients with TBI caused by RTAs. The cohort was divided into CE and non-CE groups, according to CT results within 7 days. Demographic data, imaging data, and clinical data were collected and analyzed. Quantitative variables that follow normal distribution were presented as mean ± standard deviation, those that do not follow normal distribution were presented as median (Q 1, Q 3). Categorical variables were expressed as percentages. The Chi-square test and logistic regression analysis were used to identify risk factors for CE. Logistic curve fitting was performed to predict the time to secondary CE in TBI patients with different levels of injuries. The efficacy of the model was evaluated using the receiver operator characteristic curve. Results::According to the study, almost half (47.3%) of the patients were found to have CE. The risk factors associated with CE were bilateral frontal lobe contusion, unilateral frontal lobe contusion, cerebral contusion, subarachnoid hemorrhage, and abbreviated injury scale (AIS). The odds ratio values for these factors were 7.27 (95% confidence interval ( CI): 2.08 -25.42, p = 0.002), 2.85 (95% CI: 1.11 -7.31, p = 0.030), 2.62 (95% CI: 1.12 -6.13, p = 0.027), 2.44 (95% CI: 1.25 -4.76, p = 0.009), and 1.5 (95% CI: 1.10 -2.04, p = 0.009), respectively. We also observed that patients with mild/moderate TBI (AIS ≤ 3) had a 50% probability of developing CE 19.7 h after injury (χ 2= 13.82, adjusted R2 = 0.51), while patients with severe TBI (AIS > 3) developed CE after 12.5 h (χ 2= 18.48, adjusted R2 = 0.54). Finally, we conducted a receiver operator characteristic curve analysis of CE time, which showed an area under the curve of 0.744 and 0.672 for severe and mild/moderate TBI, respectively. Conclusion::Our study found that the onset of CE in individuals with TBI resulting from RTAs was correlated with the severity of the injury. Specifically, those with more severe injuries experienced an earlier onset of CE. These findings suggest that there is a critical time window for clinical intervention in cases of CE secondary to TBI.
7.Study on the Regulatory Effect of Xinkang Granules on Inflammatory Factors in Rats with Chronic Heart Failure Based on the cGAS/STING Signaling Pathway
Siqin TANG ; Bing GUO ; Liang LI ; Qingqi YIN ; Qinliang WU ; Yilin MAO
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(5):674-680
Objective To explore the intervention effect and molecular mechanism of Xinkang Granules on inflammatory factors in rats with chronic heart failure based on cGAS/STING signaling pathway.Methods SD rats were randomly divided into normal group and modeling group.The chronic heart failure model was established by intraperitoneal injection of Doxorubicin Hydrochloride.After successfully modeling,the rats were further divided into model group,Valsartan group and Xinkang Granules group.The model group was treated with distilled water every day,the Valsartan group was treated with Valsartan solution every day,and the Xinkang Granules group was treated with Xinkang Granules every day,all given for 4 consecutive weeks.Echocardiography was used to detect cardiac function,the pathological changes of myocardium were detected by hematoxylin-eosin staining(HE),the ultrastructural changes of myocardium in each group were observed by transmission electron microscope,and the contents of interleukin-1β(IL-1β)and interleukin-6(IL-6)in serum were detected by enzyme-linked immunosorbent assay(ELISA).The mRNA expression levels of mitochondrial transcription factor A(TFAM),cyclic guanosine monophosphate-adenylate synthase(cGAS),interferon-stimulated gene(STING)and IL-6 in myocardial tissue of rats in each group were detected by real-time fluorescence quantitative method(qPCR).The protein expressions of cGAS and STING in rat myocardial tissue were detected by immunohistochemical method.Results Compared with the blank group,the rats in the model group had significant inflammatory cell infiltration and inflammatory edema in myocardial tissue,their cardiac function was significantly reduced(P<0.05,P<0.01),and serum inflammatory factors were significantly increased(P<0.01).The mRNA expression of TFAM in myocardial tissue was significantly reduced(P<0.01),the mRNA expressions of IL-6,cGAS,and STING were significantly increased(P<0.01),and the protein expressions of cGAS and STING in the myocardial tissue were significantly increased(P<0.01).Compared with the model group,the cardiac function of the rats in the Xinkang Granules group was significantly improved(P<0.05,P<0.01),the inflammatory infiltration of myocardial cells was reduced,the expression of serum inflammatory factors was significantly reduced(P<0.01),the mRNA expression of TFAM in myocardial tissue was significantly increased(P<0.05),and the mRNA expressions of IL-6,cGAS,and STING were significantly decreased(P<0.01),the protein expressions of cGAS and STING in myocardial tissue were significantly decreased(P<0.01).Conclusion Xinkang Granules can reduce the expression of inflammatory factors and improve cardiac function in rats with chronic heart failure.Its mechanism may be related to inhibiting the cGAS/STING signaling pathway.
8.Advances and challenges in using nirmatrelvir and its derivatives against SARS-CoV-2 infection
Wujun CHEN ; Bing LIANG ; Xiaolin WU ; Ling LI ; Chao WANG ; Dongming XING
Journal of Pharmaceutical Analysis 2023;13(3):255-261
On December 22,2021,the United States Food and Drug Administration approved the first main protease inhibitor,i.e.,oral antiviral nirmatrelvir(PF-07321332)/ritonavir(Paxlovid),for the treatment of early severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection.Nirmatrelvir inhibits SARS-CoV-2 infection,but high doses or long-term treatment may cause embryonic developmental toxicity and changes in host gene expression.The chiral structure of nirmatrelvir plays a key role in its antiviral activity.Ritonavir boosts the efficacy of nirmatrelvir by inactivating cytochrome P450 3A4 expression and occupying the plasma protein binding sites.Multidrug resistance protein 1 inhibitors may increase the efficacy of nirmatrelvir.However,Paxlovid has many contraindications.Some patients treated with Paxlovid experience a second round of coronavirus disease 2019(COVID-19)symptoms soon after re-covery.Interestingly,the antiviral activity of nirmatrelvir metabolites,such as compounds 12-18,is similar to or higher than that of nirmatrelvir.Herein,we review the advances and challenges in using nirmatrelvir and its derivatives with the aim of providing knowledge for drug developers and physicians in the fight against COVID-19.
9.Cytotoxicity of 4 Wild Mushrooms in a Case of Yunnan Sudden Unexplained Death.
Wu LONG ; Peng-Fei QU ; Lin MA ; Rui WANG ; Yan-Mei XI ; Yu-Hua LI ; Sheng-Jie NIE ; Ting DUAN ; Jin-Liang DU ; Xue TANG ; Jing-Feng ZHAO ; Pu-Ping LEI ; Yue-Bing WANG
Journal of Forensic Medicine 2023;39(2):121-128
OBJECTIVES:
To explore the cytotoxicity of four wild mushrooms involved in a case of Yunnan sudden unexplained death (YNSUD), to provide the experimental basis for prevention and treatment of YNSUD.
METHODS:
Four kinds of wild mushrooms that were eaten by family members in this YNSUD incident were collected and identified by expert identification and gene sequencing. Raw extracts from four wild mushrooms were extracted by ultrasonic extraction to intervene HEK293 cells, and the mushrooms with obvious cytotoxicity were screened by Cell Counting Kit-8 (CCK-8). The selected wild mushrooms were prepared into three kinds of extracts, which were raw, boiled, and boiled followed by enzymolysis. HEK293 cells were intervened with these three extracts at different concentrations. The cytotoxicity was detected by CCK-8 combined with lactate dehydrogenase (LDH) Assay Kit, and the morphological changes of HEK293 cells were observed under an inverted phase contrast microscope.
RESULTS:
Species identification indicated that the four wild mushrooms were Butyriboletus roseoflavus, Boletus edulis, Russula virescens and Amanita manginiana. Cytotoxicity was found only in Amanita manginiana. The raw extracts showed cytotoxicity at the mass concentration of 0.1 mg/mL, while the boiled extracts and the boiled followed by enzymolysis extracts showed obvious cytotoxicity at the mass concentration of 0.4 mg/mL and 0.7 mg/mL, respectively. In addition to the obvious decrease in the number of HEK293 cells, the number of synapses increased and the refraction of HEK293 cells was poor after the intervention of Amanita manginiana extracts.
CONCLUSIONS
The extracts of Amanita manginiana involved in this YNSUD case has obvious cytotoxicity, and some of its toxicity can be reduced by boiled and enzymolysis, but cannot be completely detoxicated. Therefore, the consumption of Amanita manginiana is potentially dangerous, and it may be one of the causes of the YNSUD.
Humans
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HEK293 Cells
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Sincalide
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China
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Amanita
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Death, Sudden
10.Regulation of Baicalin on Growth of Extranodal NK/T Cell Lymphoma Cells through FOXO3/CCL22 Signaling Pathway.
Xiao-Hui DUAN ; Hong LI ; Yao LYU ; Jing LIU ; Shi-Xiong WANG ; Zhen-Tian WU ; Bing-Xuan WANG ; Ming LU ; Jian-Hong WANG ; Rong LIANG
Journal of Experimental Hematology 2023;31(3):730-738
OBJECTIVE:
To investigate the effect of baicalin on the growth of extranodal NK/T cell lymphoma (ENKTCL) cells and its related mechanism.
METHODS:
Normal NK cells and human ENKTCL cells lines SNK-6 and YTS were cultured, then SNK-6 and YTS cells were treated with 5, 10, 20 μmol/L baicalin and set control. Cell proliferation and apoptosis was detected by Edu method and FCM method, respectively, and expressions of BCL-2, Bax, FOXO3 and CCL22 proteins were detected by Western blot. Interference plasmids were designed and synthesized. FOXO3 siRNA interference plasmids and CCL22 pcDNA overexpression plasmids were transfected with PEI transfection reagent. Furthermore, animal models were established for validation.
RESULTS:
In control group and 5, 10, 20 μmol/L baicalin group, the proliferation rate of SNK-6 cells was (56.17±2.96)%, (51.92±4.63)%, (36.42±1.58)%, and (14.60±2.81)%, respectively, while that of YTS cells was (58.85±2.98)%, (51.38±1.32)%, (34.75±1.09)%, and (15.45±1.10)%, respectively. In control group and 5, 10, 20 μmol/L baicalin group, the apoptosis rate of SNK-6 cells was (5.93±0.74)%, (11.78±0.34)%, (28.46±0.44)%, and (32.40±0.37)%, respectively, while that of YTS cells was (7.93±0.69)%, (16.29±1.35)%, (33.91±1.56)%, and (36.27±1.06)%, respectively. Compared with control group, the expression of BCL-2 protein both in SNK-6 and YTS cells decreased significantly (P<0.001), and the expression of Bax protein increased in SNK-6 cells only when the concentration of baicalin was 20 μmol/L (P<0.001), while that in YTS cells increased in all three concentrations(5, 10, 20 μmol/L) of baicalin (P<0.001). The expression of FOXO3 protein decreased while CCL22 protein increased in ENKTCL cell lines compared with human NK cells (P<0.001), but the expression of FOXO3 protein increased (P<0.01) and CCL22 protein decreased after baicalin treatment (P<0.001). Animal experiments showed that baicalin treatment could inhibit tumor growth. The expression of CCL22 protein in ENKTCL tissue of nude mice treated with baicalin decreased compared with control group (P<0.01), while the FOXO3 protein increased (P<0.05). In addition, FOXO3 silencing resulted in the decrease of FOXO3 protein expression and increase of CCL22 protein expression (P<0.01, P<0.001).
CONCLUSION
Baicalin can inhibit proliferation and promote apoptosis of ENKTCL cell lines SNK-6 and YTS, up-regulate the expression of Bax protein, down-regulate the expression of BCL-2 protein, and down-regulate the expression of CCL22 protein mediated by FOXO3. Animal experiment shown that the baicalin can inhibit tumor growth. Baicalin can inhibit the growth and induce apoptosis of ENKTCL cells through FOXO3/CCL22 signaling pathway.
Animals
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Mice
;
Humans
;
Lymphoma, Extranodal NK-T-Cell/pathology*
;
Forkhead Box Protein O3/metabolism*
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bcl-2-Associated X Protein/pharmacology*
;
Mice, Nude
;
Signal Transduction
;
Apoptosis
;
Proto-Oncogene Proteins c-bcl-2/metabolism*
;
Chemokine CCL22/pharmacology*


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