BACKGROUND:A large number of studies have shown that neurodegenerative diseases are closely related to oxidative stress injury and the imbalance of mitochondrial dynamics.Lycium barbarum polysaccharides have a neuroprotective effect.However,it is not clear whether lycium barbarum polysaccharides can ameliorate apoptosis induced by oxidative stress injury by regulating abnormal mitochondrial dynamics.OBJECTIVE:To study the effect of lycium barbarum polysaccharides on apoptosis induced by H2O2 in SH-SY5Y human neuroblastoma cells.METHODS:SH-SY5Y cells were cultured in three groups.The control group was cultured for 24 hours.The hydrogen peroxide group was treated with H2O2 for 24 hours,and the lycium barbarum polysaccharide group was treated with lycium barbarum polysaccharide for 2 hours and then treated with H2O2 for 24 hours.After treatment,the levels of malondialdehyde,glutathione,and superoxide dismutase in the precipitation of the cells were detected by kit.Mitochondrial membrane potential was detected by JC-1 kit.Cell viability was detected by MTT assay.Apoptosis was detected by TUNEL.The expression levels of mitochondrial dynamics-related proteins (phosphorylated promoter protein 1,mitochondrial fission protein 1,mitochondrial fusion protein 1,mitochondrial fusion protein 2,and optic atrophy protein 1) and apoptotic proteins (Bax,Bcl-2,and Caspase-3) were detected by immunofluorescence staining and western blot assay.RESULTS AND CONCLUSION:(1) Compared with the control group,the levels of malondialdehyde were increased (P<0.05),and the levels of superoxide dismutase and glutathione were decreased (P<0.05) in the H2O2 group.Compared with the H2O2 group,the malondialdehyde level was decreased (P<0.05),and the superoxide dismutase and glutathione levels were increased (P<0.05) in the lycium barbarum polysaccharide group.(2) The mitochondrial membrane potential in the H2O2 group was lower than that in the control group (P<0.05),and that of lycium barbarum polysaccharide group was higher than that of the H2O2 group (P<0.05).(3) Compared with the control group,the apoptosis rate and the expression of Bax and Caspase-3 protein were increased (P<0.05),while the cell viability and the expression of Bcl-2 protein were decreased (P<0.05) in the H2O2 group.Compared with the H2O2 group,the apoptosis rate and the expression of Bax and Caspase-3 protein were decreased (P<0.05),while the cell viability and the expression of Bcl-2 protein were increased (P<0.05) in the lycium barbarum polysaccharide group.(4) Compared with the control group,the protein expression levels of phosphorylated promoter protein 1 and mitochondrial fission protein 1 were increased (P<0.05),and the protein expression levels of mitochondrial fusion protein 1,mitochondrial fusion protein 2,and optic atrophy protein 1 were decreased (P<0.05) in the H2O2 group.Compared with the H2O2 group,the protein expression levels of phosphorylated promoter protein 1 and mitochondrial fission protein 1 were decreased (P<0.05),and the protein expression levels of mitochondrial fusion protein 1,mitochondrial fusion protein 2,and optic atrophy protein 1 were increased (P<0.05) in the lycium barbarum polysaccharide group.(5) These results indicate that lycium barbarum polysaccharide can improve SH-SY5Y cell apoptosis caused by oxidative stress damage by regulating mitochondrial dynamics.

Objective:To investigate the clinical features, diagnostic and therapeutic methods, and prognosis of patients with acute myeloid leukemia (AML) who are positive for multiple genes.Methods:The clinical data of a 35-year-old male AML patient, who was admitted to The 940 th Hospital of the PLA Joint Logistics Support Force on October 10, 2021, were analyzed based on related literature. Results:The patient was admitted due to rhinorrhea for 1 month and sore throat with cervical lymph node enlargement for 3 days. Based on laboratory test results, the patient was diagnosed with acute myelomonocytic leukemia with eosinophilia and mutations in the core-binding factor β subunit gene ( CBFβ-MYH11), located on the long arm of chromosome 16, as well as mutations in the Wilms' tumor 1 gene ( WT1), the stem cell factor receptor gene ( KIT), and the neuroblastoma RAS viral oncogene homolog ( NRAS). The patient was treated multiple times with a regimen of idarubicin combined with cytarabine (Ara-C), achieving complete remission with negative minimal residual disease detection. The patient tolerated the treatment well. However, after the fourth cycle of chemotherapy with Ara-C, the patient developed grade Ⅳ bone marrow suppression. Following treatment to increase white blood cells and platelets, as well as blood component transfusions, the patient's condition improved. Subsequently, after receiving intensified Ara-C treatment, the patient's condition stabilized. Conclusions:AML with a positive CBFβ- MYH11 fusion gene has a favorable prognosis. However, the presence of a concomitant KIT mutation and a high proportion of primitive white blood cells in the peripheral blood at initial diagnosis may affect the prognosis of this type of AML. Additionally, WT1 is an independent prognostic factor for AML. RAS gene mutations do not impact overall survival, disease-free survival, complete remission, or relapse rates.

Objective:To investigate the effect and mechanism of lycium barbarum polysaccharide(LBP)on lipopolysaccharide(LPS)-induced inflammatory response of NLRP3 inflammasome in BV2 microglial cells.Methods:BV2 microglial cells were routinely cultured.CCK-8 assay was used to detect the effect of different concentrations(0.5,1,1.5,2 g/L)LBP on cell activity.Cells were di-vided into three groups:control group,LPS group and LBP+LPS group.Effect of LBP on LPS-induced cell activity was detected by CCK-8 method;RT-qPCR and immunofluorescence assay were used to detect NLRP3,ASC,Caspase-1,IL-18 and IL-1β expressions.Western blot was used to detect expressions of NLRP3,ASC,Caspase-1,TLR4,MyD88,NF-κB p65,IL-18,IL-1β and TNF-α pro-teins.Results:CCK-8 assay showed that 1 g/L LBP was the most applicable.Compared with control group,cell viability in LPS group was decreased;RT-qPCR,immunofluorescence and Western blot results showed that fluorescence intensity,mRNA and protein expres-sions of NLRP3,ASC,Caspase-1,IL-18 and IL-1β were increased in LPS group.Western blot results showed that TLR4,MyD88,NF-κB p65 and TNF-α protein expressions were increased in LPS group.After LBP treatment,cell viability was increased;expres-sions of NLRP3,ASC,Caspase-1,NF-κB p65,TLR4,MyD88,IL-18,IL-1β and TNF-α were decreased.Conclusion:LBP may in-hibit LPS-induced NLRP3 inflammatory vesicles in BV2 cells via TLR4/MyD88/NF-κB signaling pathway.

ObjectiveTo establish a model that can quickly identify the aroma components in different parts of Nardostachys jatamansi， so as to provide a quality control basis for the market circulation and clinical use of N. jatamansi. MethodsHeadspace solid-phase microextraction-gas chromatography-mass spectrometry（HS-SPME-GC-MS） combined with Smart aroma database and National Institute of Standards and Technology（NIST） database were used to characterize the aroma components in different parts of N. jatamansi， and the aroma components were quantified according to relative response factor（RRF） and three internal standards， and the markers of aroma differences in different parts of N. jatamansi were identified by orthogonal partial least squares-discriminant analysis（OPLS-DA） and cluster thermal analysis based on variable importance in the projection（VIP） value >1 and P<0.01. The odor data of different parts of N. jatamansi were collected by Heracles Ⅱ Neo ultra-fast gas phase electronic nose， and the correlation between compound types of aroma components collected by the ultra-fast gas phase electronic nose and the detection results of HS-SPME-GC-MS was investigated by drawing odor fingerprints and odor response radargrams. Chromatographic peak information with distinguishing ability≥0.700 and peak area≥200 was selected as sensor data， and the rapid identification model of different parts of N. jatamansi was established by principal component analysis（PCA）， discriminant factor alysis（DFA）， soft independent modeling of class analogies（SIMCA） and statistical quality control analysis（SQCA）. ResultsThe HS-SPME-GC-MS results showed that there were 28 common components in the underground and aboveground parts of N. jatamansi， of which 22 could be quantified and 12 significantly different components were screened out. Among these 12 components， the contents of five components（ethyl isovalerate， 2-pentylfuran， benzyl alcohol， nonanal and glacial acetic acid，） in the aboveground part of N. jatamansi were significantly higher than those in the underground part（P<0.01）， the contents of β-ionone， patchouli alcohol， α-caryophyllene， linalyl butyrate， valencene， 1，8-cineole and p-cymene in the underground part of N. jatamansi were significantly higher than those in the aboveground part（P<0.01）. Heracles Ⅱ Neo electronic nose results showed that the PCA discrimination index of the underground and aboveground parts of N. jatamansi was 82， and the contribution rates of the principal component factors were 99.94% and 99.89% when 2 and 3 principal components were extracted， respectively. The contribution rate of the discriminant factor 1 of the DFA model constructed on the basis of PCA was 100%， the validation score of the SIMCA model for discrimination of the two parts was 99， and SQCA could clearly distinguish different parts of N. jatamansi. ConclusionHS-SPME-GC-MS can clarify the differential markers of underground and aboveground parts of N. jatamansi. The four analytical models provided by Heracles Ⅱ Neo electronic nose（PCA， DFA， SIMCA and SQCA） can realize the rapid identification of different parts of N. jatamansi. Combining the two results， it is speculated that terpenes and carboxylic acids may be the main factors contributing to the difference in aroma between the underground and aboveground parts of N. jatamansi.

BACKGROUND:A large number of studies have shown that neurodegenerative diseases are closely related to oxidative stress injury and the imbalance of mitochondrial dynamics.Lycium barbarum polysaccharides have a neuroprotective effect.However,it is not clear whether lycium barbarum polysaccharides can ameliorate apoptosis induced by oxidative stress injury by regulating abnormal mitochondrial dynamics.OBJECTIVE:To study the effect of lycium barbarum polysaccharides on apoptosis induced by H2O2 in SH-SY5Y human neuroblastoma cells.METHODS:SH-SY5Y cells were cultured in three groups.The control group was cultured for 24 hours.The hydrogen peroxide group was treated with H2O2 for 24 hours,and the lycium barbarum polysaccharide group was treated with lycium barbarum polysaccharide for 2 hours and then treated with H2O2 for 24 hours.After treatment,the levels of malondialdehyde,glutathione,and superoxide dismutase in the precipitation of the cells were detected by kit.Mitochondrial membrane potential was detected by JC-1 kit.Cell viability was detected by MTT assay.Apoptosis was detected by TUNEL.The expression levels of mitochondrial dynamics-related proteins (phosphorylated promoter protein 1,mitochondrial fission protein 1,mitochondrial fusion protein 1,mitochondrial fusion protein 2,and optic atrophy protein 1) and apoptotic proteins (Bax,Bcl-2,and Caspase-3) were detected by immunofluorescence staining and western blot assay.RESULTS AND CONCLUSION:(1) Compared with the control group,the levels of malondialdehyde were increased (P<0.05),and the levels of superoxide dismutase and glutathione were decreased (P<0.05) in the H2O2 group.Compared with the H2O2 group,the malondialdehyde level was decreased (P<0.05),and the superoxide dismutase and glutathione levels were increased (P<0.05) in the lycium barbarum polysaccharide group.(2) The mitochondrial membrane potential in the H2O2 group was lower than that in the control group (P<0.05),and that of lycium barbarum polysaccharide group was higher than that of the H2O2 group (P<0.05).(3) Compared with the control group,the apoptosis rate and the expression of Bax and Caspase-3 protein were increased (P<0.05),while the cell viability and the expression of Bcl-2 protein were decreased (P<0.05) in the H2O2 group.Compared with the H2O2 group,the apoptosis rate and the expression of Bax and Caspase-3 protein were decreased (P<0.05),while the cell viability and the expression of Bcl-2 protein were increased (P<0.05) in the lycium barbarum polysaccharide group.(4) Compared with the control group,the protein expression levels of phosphorylated promoter protein 1 and mitochondrial fission protein 1 were increased (P<0.05),and the protein expression levels of mitochondrial fusion protein 1,mitochondrial fusion protein 2,and optic atrophy protein 1 were decreased (P<0.05) in the H2O2 group.Compared with the H2O2 group,the protein expression levels of phosphorylated promoter protein 1 and mitochondrial fission protein 1 were decreased (P<0.05),and the protein expression levels of mitochondrial fusion protein 1,mitochondrial fusion protein 2,and optic atrophy protein 1 were increased (P<0.05) in the lycium barbarum polysaccharide group.(5) These results indicate that lycium barbarum polysaccharide can improve SH-SY5Y cell apoptosis caused by oxidative stress damage by regulating mitochondrial dynamics.

Objective:To investigate the effect and mechanism of lycium barbarum polysaccharide(LBP)on lipopolysaccharide(LPS)-induced inflammatory response of NLRP3 inflammasome in BV2 microglial cells.Methods:BV2 microglial cells were routinely cultured.CCK-8 assay was used to detect the effect of different concentrations(0.5,1,1.5,2 g/L)LBP on cell activity.Cells were di-vided into three groups:control group,LPS group and LBP+LPS group.Effect of LBP on LPS-induced cell activity was detected by CCK-8 method;RT-qPCR and immunofluorescence assay were used to detect NLRP3,ASC,Caspase-1,IL-18 and IL-1β expressions.Western blot was used to detect expressions of NLRP3,ASC,Caspase-1,TLR4,MyD88,NF-κB p65,IL-18,IL-1β and TNF-α pro-teins.Results:CCK-8 assay showed that 1 g/L LBP was the most applicable.Compared with control group,cell viability in LPS group was decreased;RT-qPCR,immunofluorescence and Western blot results showed that fluorescence intensity,mRNA and protein expres-sions of NLRP3,ASC,Caspase-1,IL-18 and IL-1β were increased in LPS group.Western blot results showed that TLR4,MyD88,NF-κB p65 and TNF-α protein expressions were increased in LPS group.After LBP treatment,cell viability was increased;expres-sions of NLRP3,ASC,Caspase-1,NF-κB p65,TLR4,MyD88,IL-18,IL-1β and TNF-α were decreased.Conclusion:LBP may in-hibit LPS-induced NLRP3 inflammatory vesicles in BV2 cells via TLR4/MyD88/NF-κB signaling pathway.

Objective:To investigate the clinical features, diagnostic and therapeutic methods, and prognosis of patients with acute myeloid leukemia (AML) who are positive for multiple genes.Methods:The clinical data of a 35-year-old male AML patient, who was admitted to The 940 th Hospital of the PLA Joint Logistics Support Force on October 10, 2021, were analyzed based on related literature. Results:The patient was admitted due to rhinorrhea for 1 month and sore throat with cervical lymph node enlargement for 3 days. Based on laboratory test results, the patient was diagnosed with acute myelomonocytic leukemia with eosinophilia and mutations in the core-binding factor β subunit gene ( CBFβ-MYH11), located on the long arm of chromosome 16, as well as mutations in the Wilms' tumor 1 gene ( WT1), the stem cell factor receptor gene ( KIT), and the neuroblastoma RAS viral oncogene homolog ( NRAS). The patient was treated multiple times with a regimen of idarubicin combined with cytarabine (Ara-C), achieving complete remission with negative minimal residual disease detection. The patient tolerated the treatment well. However, after the fourth cycle of chemotherapy with Ara-C, the patient developed grade Ⅳ bone marrow suppression. Following treatment to increase white blood cells and platelets, as well as blood component transfusions, the patient's condition improved. Subsequently, after receiving intensified Ara-C treatment, the patient's condition stabilized. Conclusions:AML with a positive CBFβ- MYH11 fusion gene has a favorable prognosis. However, the presence of a concomitant KIT mutation and a high proportion of primitive white blood cells in the peripheral blood at initial diagnosis may affect the prognosis of this type of AML. Additionally, WT1 is an independent prognostic factor for AML. RAS gene mutations do not impact overall survival, disease-free survival, complete remission, or relapse rates.

Objective:To observe and evaluate the clinical efficacy and safety of albumin-bound paclitaxel as first-line treatment for patients with advanced pancreatic cancer in China, and to explore the prognosis-related molecules in pancreatic cancer based on next-generation sequencing (NGS) of tumor tissues.Methods:From December 2018 to December 2020, patients with locally advanced or metastatic pancreatic cancer were recruited to accept albumin-bound paclitaxel as first-line treatment in the oncology departments of 24 hospitals in East China. The primary endpoints were overall survival (OS) and treatment related adverse events, and the secondary endpoint was progression-free survival (PFS). Adverse effects were graded using Common Terminology Criteria for Adverse Events 5.0 (CTCAE 5.0). NGS sequencing on the primary or metastatic tissue samples of pancreatic cancer obtained through surgical resection or biopsy was performed.Results:This study recruited 229 patients, including 70 patients with locally advanced pancreatic cancer (LAPC) and 159 patients with metastatic pancreatic cancer (mPC). The disease control rate was 79.9% and the objective response rate is 36.3%.The common adverse effects during treatment were anaemia (159 cases), leucopenia (170 cases), neutropenia (169 cases), increased aminotransferases (110 cases), and thrombocytopenia (95 cases), and the incidence of grade 3-4 neutropenia is 12.2% (28/229). The median follow-up time was 21.2 months (95% CI: 18.5-23.1 months). The median PFS (mPFS) was 5.3 months (95% CI: 4.37-4.07 months) and the median OS (mOS) was 11.2 months (95% CI: 9.5-12.9 months). The mPFS of patients with LAPC was 7.4 months (95% CI: 6.6-11.2 months), and their mOS was 15.5 months (95% CI: 12.6-NA months). The mPFS of patients with mPC was 3.9 months (95% CI: 3.4-5.1 months), and their mOS was 9.3 months (95% CI: 8.0-10.8 months). Multivariate Cox regression analysis showed that clinical stage ( HR=1.47, 95% CI: 1.06-2.04), primary tumor site ( HR=0.64, 95% CI: 0.48-0.86), Eastern Cooperative Oncology Group Performance Status (ECOG PS) score ( HR=2.66, 95% CI: 1.53-4.65), and whether to combine radiotherapy ( HR=0.65, 95% CI: 0.42-1.00) were independent influencing factors for the PFS of these patients. The primary tumor site ( HR=0.68, 95% CI: 0.48-0.95), ECOG score ( HR=5.82, 95% CI: 3.14-10.82), and whether to combine radiotherapy ( HR=0.58, 95% CI: 0.35-0.96) were independent influencing factors of the OS of these patients. The most frequent gene mutations in these advanced stage pancreatic patients were KRAS (89.66%), TP53 (77.01%), CDKN2A (32.18%), and SMAD4 (21.84%) by NGS of tumor tissues from 87 pancreatic cancer patients with sufficient specimens. Further analysis revealed that mutations in CDKN2B, PTEN, FGF6, and RBBP8 genes were significantly associated with an increased risk of death ( P＜0.05). Conclusion:Albumin-bound paclitaxel as first-line treatment demonstrated feasible anti-tumor efficacy and manageable safety for patients with advanced pancreatic cancer in China.

Objective To explore the feasibility of automatic segmentation of clinical target volume(CTV)and organs at risk(OARs)for cervical cancer using AccuLearning(AL)based on geometric and dosimetric indices.Methods Seventy-five CT localization images with manual contouring data of postoperative cervical cancer were enrolled in this study.Sixty cases were randomly selected to trained to generate automatic segmentation model by AL,and the CTV and OARs of the remaining 15 cases were automatically contoured.Radiotherapy plans on the automatic segmentation contours were imported on the CT images of manual contours.The efficiency,Dice similarity coefficient(DSC),Hausdorff distance(HD)and dosimetric parameters were compared between the two methods.Results The time of automatic segmentation was significantly shorter than that of the manual contour(P<0.05).The DSC of all structures were≥0.87.The HD of bowel bag and rectum were about 10 mm,and that of the rest of OARs were less than 5 mm.CTV(D98,V90% ,V95% ,Dmean,HI),bowel bag(V50)and bladder(V50)had significant differences in dosimetric comparison(P<0.05).Conclusion The automatic segmentation model based on AL can improve the efficiency of radiotherapy.Automatic segmentation of OARs has the potential of clinical application,while that of CTV still needs to be further modified.

Objective To explore the correlation between intestinal dose and acute radiation enteritis(ARE)in patients with cervical cancer received concurrent chemoradiotherapy,and optimize the dose limit of intestinal tissue.Methods 158 cervical cancer patients received concurrent chemoradiotherapy from 2014 to 2019 were selected in this study.According to CTCAE 5.0,patients with ARE≥grade 2 were classified as ARE≥grade 2 group,otherwise classified as ARE0.7,P<0.05).Conclusions For patients with cervical cancer received concurrent chemoradiotherapy,the dose of bowelbag V5 and V40 should be considered to rationally optimize the dose of bowelbag in the radiotherapy plan,so as to reduce the incidence of ARE≥grade 2.