Malignant tumor metastasis is the key factor leading to poor prognosis of patients， and it is a difficult problem to be overcome in the field of tumor therapy. Metabolic reprogramming， as a key link in the regulation of tumor metastasis activity， affects the growth， invasion， and metastasis of tumor cells by changing the metabolic pathways of intracellular substances （such as glucose， amino acids， lipids， and nucleotides）. In particular， metabolic reprogramming plays a key role in the multistage linked steps related to tumor metastasis and can play a crucial role in several key stages of tumor tissue dissociation in situ， hematogenous metastasis， and remote colonization. Malignant tumor cells can selectively adjust their own metabolic state to adapt to the growth conditions of different metastatic microenvironments and colonization sites and then choose the most favorable growth and metabolism strategy. According to the holistic concept of traditional Chinese medicine （TCM）， the metastasis of malignant tumors is generally closely related to the metabolic state of the whole body. One of the advantages of TCM in the treatment of malignant tumors is systemic regulation. With its multi-pathway， multi-target， and multi-component therapeutic characteristics， TCM can effectively control the metastasis of malignant tumors by regulating the degradation of tumor epithelial mesenchymal transformation （EMT） and extracellular matrix （ECM）， anchoring the independent growth of tumor cells and the tumor microenvironment. In this paper， the potential regulatory effects of metabolic reprogramming on the metastasis of malignant tumors were discussed， and the latest research progress of the regulation of metabolic reprogramming by TCM on tumor metastasis was reviewed. At the same time， the key targets of TCM and its bioactive components in the process of tumor metastasis intervention were reviewed. This study aims to provide a more valuable basis and clearer idea for the treatment of malignant tumor metastasis by regulating metabolic reprogramming with TCM.

Objective To investigate the role and possible mechanisms of transactive response DNA binding protein 43 (TDP-43)in mediating neuronal injury induced by oxidative stress in mouse neuro-2a (N2a)cells and mouse pain sensitization.Methods ①To evaluate the optimal induction concentration,N2a cells were treated with different concentrations of H2O2,and the cells were divided into control group,200,400 and 800 μmol/L H2O2 groups.②To assess the optimal induction duration,N2a cells were treated with 400 μmol/L H2O2,and the cells were divided into control group,and the cell groups treated for 6,12 and 24 h,respectively.③To validate the mitochondrial DNA (mtDNA)release pathway,cyclosporin A (CsA) was used to inhibit the mitochondrial permeability transition pore (mPTP),and the cells were divided into control group,24 h H2O2 group and 24 h H2O2+CsA group.④To validate TDP-43-mediated cellular damage,the cells were divided into control group,24 h H2O2 group and 24 h H2O2+siTDP-43 group.⑤Cell viability was assessed using CCK-8 assay,while cell proliferation was determined using EdU assay.Western blot analysis was employed to examine the expression levels of TDP-43,neuronal nuclei (NeuN),cyclic GMP-AMP synthase (cGAS),and stimulator of interferon genes (STING).qPCR was utilized to measure the release of mtDNA.Immunostaining was conducted to observe intracellular expression of TDP-43,and Calcein AM staining was employed to evaluate mPTP opening status.⑥To elucidate the role of TDP-43 in neuropathic pain (NP),24 healthy SPF male C57BL/6J mice (6～8 weeks old,25～30 g)were randomly divided into control group,chronic constriction injury (CCI)group,and CCI+siTDP-43 group.In 1 d before and 7,14 and 21 d after surgery,intrathecal injections of siTDP-43 were administered.Mechanical and thermal pain thresholds of the mice were assessed using von Frey filaments and radiant heat,respectively,on 1 preoperatively and 1,3,5,7,14 and 21 d postoperatively.Immunofluorescence assay was conducted on 21 d postoperatively to examine the changes in TDP-43 and NeuN in the lumbar spinal dorsal horn (L5-L6).Results Oxidative stress induced a significant increase in TDP-43 protein level in N2a cells,prompted mtDNA release through mPTP,markedly up-regulated the expression of cGAS and STING,and consequently impacted the viability of N2a cells (P<0.05).CsA treatment inhibited mPTP channel opening and thus effectively blocked mtDNA release (P<0.05),down-regulated TDP-43 and thus significantly reduced mtDNA release,suppressed the expression of cGAS and STING,and finally restored the proliferation ability of N2a cells (P<0.05).The mechanical and thermal pain thresholds exhibited a significant decrease since 5 d after CCI,which then persisted until 21 d (P<0.05).The expression of TDP-43 in spinal cord dorsal horn neurons was increased in the mice in 21 d after CCI (P<0.05),and intrathecal injection of siRNA inhibited TDP-43 expression and effectively increased the mechanical and thermal pain thresholds in the CCI mice (P<0.05).Conclusion Oxidative stress induces an up-regulation in TDP-43 protein in neurons,which stimulates the release of mtDNA into the cytoplasm through mPTP,and subsequently activates the cGAS/STING pathway,and finally,results in neuronal injury and pain sensitization in CCI mice.

Objective To investigate the effects of damaged nerve-derived mitochondrial DNA (mtDNA)on GRP75 protein expression,mitochondrial Ca2+level and apoptosis in human glioblastoma cell line U-87MG (U87)cells,and its mechanism of inducing pain sensitization in mice.Methods Chronic constriction injury (CCI) of the sciatic nerve was used to establish a mouse model of neuropathic pain (NeP).A total of 20 male C57BL/6 mice (6～8 weeks old,weighing 20～30 g)were randomly divided into sham group,and CCI-7,-14 and CCI-21 d groups.Paw withdrawal mechanical threshold (PWMT)was measured,and the changes in mtDNA content in the spinal cord were detected with quantitative real-time PCR (qPCR).After mtDNA was extracted from human neuroblastoma cells (SH-SY5Y)treated with H2O2,U87 cells were treated with mtDNA at a dose of 0～1 ng/μL.CCK-8 assay was utilized to detect cell viability,and the appropriate concentration was selected according to the results.Then U87 cells were divided into:control group,mtDNA group,mtDNA+GRP75 inhibitor (MKT-0771 μg/mL)group,mtDNA+calcium chelator (BAPTA-AM10 μmol/L)group,and mtDNA+MKT-077+BAPTA-AM group.MKT-077 and BAPTA-AM were pre-treated in 2 h before mtDNA treatment,respectively.Western blotting was employed to assess the expression of glucose-regulated protein 75 (GRP75) protein,and endoplasmic reticulum-mitochondrial colocalization staining was conducted to observe the endoplasmic reticulum-mitochondrial junction.Calcium ion fluorescent probe was used to measure mitochondrial Ca2+levels.The oxidative stress and function of mitochondria were evaluated by reactive oxygen species (ROS)and mitochondrial membrane potential.PI fluorescent labeling was employed to examine apoptotic rate of U87 cells.Results Compared with the sham group,the cytochrome C oxidase I (CO1)and nicotinamide adenine dinucleotide dehydrogenase 1 (ND1),which were utilized to measure mtDNA content in the spinal cord of mice in the CCI-21 group,were increased (1.01±0.20 vs 2.22±0.26,P<0.05,1.00±0.12 vs 1.79±0.07,P<0.05).CCK-8 assay showed that mtDNA (1 ng/μL) inhibited U87 cell viability (P<0.01).mtDNA (0.2 ng/μL) up-regulated the expression of GRP75 protein (P<0.05),promoted endoplasmic reticulum-mitochondrial coupling,and consequently increased mitochondrial Ca2+level (109.4±62.6 vs 540.3±150.3,P<0.05)and production of ROS (P<0.05).After MKT-077 or/and BAPTA-AM treatment,the level of mitochondrial Ca2+was decreased,the mitochondrial membrane potential was improved,and the apoptotic rate of U87 cells was reduced by PI assays (mtDNA vs mtDNA+MKT-077:18.39±2.09 vs 13.22±1.42,P<0.05;mtDNA vs mtDNA+BAPTA-AM:18.39±2.09 vs 12.09±1.53,P<0.05;mtDNA vs mtDNA+MKT-077+BAPTA-AM:18.39±2.09 vs 11.65±2.09,P<0.05).Conclusion Damaged nerve-derived mtDNA can up-regulate the expression of GRP75,interfere with mitochondrial Ca2+level and exacerbate mitochondrial oxidative stress to induce apoptosis in U87 cells,which may be a novel mechanism involved in NeP formation.

Humans ; Asthma/drug therapy* ; Biological Therapy

BACKGROUND: The switch/sucrose nonfermentable chromatin-remodeling (SWI/SNF) complex is a pivotal chromatin remodeling complex, and the genomic alterations (GAs) of the SWI/SNF complex are observed in several cancer types, correlating with multiple biological features of tumor cells. However, their role in liver metastasis of non-small cell lung cancer (NSCLC) remains unclear. Our study aims to investigate the role and potential mechanisms underlying NSCLC liver metastasis induced by the GAs of SWI/SNF complex. METHODS: The GAs of SWI/SNF complex in NSCLC cell lines (H1299, H23 and H460) were identified by whole-exome sequencing (WES). ARID1A knockout H1299 cell was constructed with the CRISPR/Cas9 technology. The mouse model of liver metastasis from NSCLC was established to simulate lung cancer liver metastasis and observe the metastasis rate under different gene mutation conditions. RNA sequencing and Western blot were conducted for differential gene expression analysis. Immunohistochemistry (IHC) analysis was used to assess protein expression levels of SWI/SNF-regulated target molecules in mouse liver metastases. RESULTS: WES analysis revealed intracellular gene mutations. The animal experiments demonstrated a correlation between the GAs of SWI/SNF complex and a higher liver metastasis rate in immunodeficient mice. Transcriptome sequencing and Western blot analysis showed upregulated expression of ALDH1A1 and APOBEC3B in SWI/SNF-mut cells, particularly in ARID1A-deficient H460 and H1299 sgARID1A cells. IHC staining of mouse liver metastases further demonstrated elevated expression of ALDH1A1 in the H460 and H1299 sgARID1A group. CONCLUSIONS This study underscores the critical role of the GAs of SWI/SNF complex, such as ARID1A and SMARCA4, in promoting liver metastasis of lung cancer cells. The GAs of SWI/SNF complex may promote liver-specific metastasis by upregulating ALDH1A1 and APOBEC3B expression, providing novel insights into the molecular mechanisms underlying lung cancer liver metastasis.
Animals ; Mice ; Carcinoma, Non-Small-Cell Lung/genetics* ; Lung Neoplasms/genetics* ; Mutation ; Liver Neoplasms/genetics*

Objective:To investigate the feasibility of applying the ArcherQA three-dimensional (3D) dosimetric verification system in intensity-modulated radiotherapy (IMRT) plans for nasopharyngeal carcinoma (NPC).Methods:A retrospective analysis was conducted for 105 NPC patients′ IMRT plans developed using the Eclipse treatment planning system (TPS). Dose verification was conducted using the ArcherQA system and through portal dosimetry (PD). Moreover, this study compared γ passing rates (criteria: 3 mm/3%, TH = 10%) between ArcherQA and PD and the doses delivered to the target volume ( Dmean, D90%) and organs at risk (OARs) ( Dmean) between ArcherQA and TPS, and analyzed the 3D γ passing rates of each organ at risk calculated by ArcherQA. Results:The average 3D γ passing rate calculated by ArcherQA was (99.04±1.01)%, and the average 2D γ passing rate measured by PD was (99.49±0.78)%, with statistically significant differences ( t=-3.35, P< 0.05). The dosimetric differences to the target volume between ArcherQA and TPS were as follows: the average difference in Dmean to the gross tumor volume (GTV) was (0.57±0.48)%, and the average difference in D90% was (0.65±0.56)%. For the target volume, the average γ passing rate was (97.67±3.43)% for GTV, (97.80±4.35)% for GTVnd-L, (97.82±4.07)% for GTVnd-R, (97.88±2.44)% for CTV1, and (96.64±4.32)% for CTV2. The mean dose difference of each target volume was CTV1 (0.57±0.46)%, GTVnd-L (0.85±0.55)%, GTVnd-R (0.73±0.55)%, and CTV2 (0.88±0.52)%. For OARs, the mean γ passing rate was (99.93±0.22)% for the brainstem, (99.17±2.82)% for the optic chiasm, (100±0)% for the lens, (99.56±1.05)% for the spinal cord, (99.00±2.06)% for the thyroid, and (87.86±10.42)% for the trachea. Statistically significant differences in the average doses to OARs were observed ( t=-14.62 to 4.82, P<0.05), except for those to the left optic nerve, the right hippocampus, and the right parotid gland. Conclusions:Based on the high-performance GPU platform and the Monte Carlo dose algorithm, ArcherQA can provide accurate 3D dose distribution and 3D γ passing rates inside patients according to CT images and provide the dose volume histogram (DVH) of various regions of interest (ROIs). Therefore, the ArcherQA three-dimensional dose verification system can be applied to IMRT plans for NPC. Moreover, it is inducive to improve the treatment efficiency since it does not occupy the accelerator operation time.

Objective:To explore the characteristics of T lymphocyte subsets and cytokines in hyperlipidemia-induced acute pancreatitis (HLAP) and its prognostic value.Methods:This study included 184 patients with acute pancreatitis (AP) admitted to the First Affiliated Hospital of Xiamen University from January 2018 to May 2021. Based on disease etiology, there were 92 HLAP cases and 92 non-hyperlipidemia-induced AP (NHLAP) cases. Stratified by disease severity according to 2012 Atlanta classification criteria, the patients were divided into the severe subgroup (SAP) and non-severe subgroup (NSAP). Peripheral venous blood samples were taken from all patients on day 1, 3, and 5 after admission. T lymphocyte subsets were determined by flow cytometry, and cytokines were detected by flow fluorometry. The number of CD4 +% and CD8 +% and the expression of cytokines were compared by Student’s t test or Mann-Whitney U analysis. Logistic regression analyses were performed to identify risk factors for severe AP, and a receiver operating characteristic (ROC) curve was constructed to predict severe AP. Statistical significance was taken as P<0.05. Results:Compared with the NHLAP group, patients in the HLAP group had lower CD4 +%, while higher levels of IL-2 on day 1 ( P<0.05), and had also lower CD4 +%, while higher levels of IL-4, IL-6, and IL-10 on day 3 ( P<0.05). Furthermore, IL-6 and IL-10 levels of the HLAP group were significantly increased compared to the NHLAP group on day 5 ( P<0.05). IL-10 levels in the SAP subgroup were significantly higher than those in the NSAP subgroup on day 1 ( P<0.05). Compared with the NSAP subgroup, the SAP subgroup had elevated levels of IL-2, IL-4, IL-6, IL-10 and IFN-γ on day 3 (all P<0.05), and had lower CD4 +%, while increased levels of IL-6 and IL-10 on day 5 (all P<0.05). Multivariate Logistic regression analysis showed that IL-10 was an immune indicator of independent risk factor for severe AP in the HLAP group on day 1 ( OR=1.139, 95% CI: 1.038-1.251, P<0.05). Finally, ROC analysis showed that the area under the curve of IL-10 to assess HLAP with severe AP was 0.772, and the best cut-off value for predicting severe AP was 5.6 pg/mL, with a sensitivity of 83.3% and a specificity of 68.8%. Conclusions:Changes of CD4 +% and cytokines are different between the HLAP and NHLAP groups. IL-10 can be used as a predictor of early disease severity in patients with HLAP.

【Objective】 To explore the relationship of histone deacetylase 4 (HDAC4) gene rs1108519 and rs3791398 polymorphisms with essential hypertension. 【Methods】 Totally 212 patients with essential hypertension were selected, and 212 healthy people were matched as the control group according to the principle of the same sex and age of ±2 years. Sanger sequencing was used to determine the genotype. SPSS22.0 software was used to compare and analyze the relationship of clinical data and gene polymorphisms with essential hypertension. 【Results】 ① Compared with those in the control group, age, BMI, pulse pressure difference, mean arterial pressure, uric acid, triglyceride, RV5, and RV5+SV1 were higher, heart rate was faster, and HDL was lower in hypertension group (all P<0.05). There was no significant difference in the other clinical data (all P>0.05). ② The genotype distribution of the two loci in the two groups were in accordance with Hardy-Weinberg equilibrium (all P>0.05). Compared with the control group, hypertension group had lower frequencies of genotype AA and allele A in rs1108519 locus, but higher frequencies of genotype AG, GG and allele G (all P<0.05). There was no significant difference in genotype or allele frequency distribution between the two groups at rs3791398 locus (all P>0.05). ③ After adjusting for possible confounding, the risk of hypertension in the population with AG and GG genotypes in rs1108519 locus was 5.980 times (OR=5.980, 95% CI: 1.334-26.811, P=0.019) and 2.832 times (OR=2.832, 95% CI: 1.466-5.472, P=0.002) that of AA genotype, respectively. Under the dominant model, the risk of hypertension in subjects with AG or GG genotype was 3.207 times higher than that with AA genotype (OR=3.207, 95% CI: 1.739-5.917, P<0.001). Under the recessive model, the risk of hypertension in subjects with AG or AA genotype was 19.3% of that with GG genotype (OR=0.193, 95% CI: 0.045-0.825, P=0.026). The risk of hypertension in subjects with allele G was 1.816 times higher than that with allele A (OR=1.816, 95% CI: 1.281-2.575, P=0.001). There was no significant difference in the risk of hypertension among people with different genotypes and alleles at rs3791398 locus (all P>0.05). 【Conclusion】 HDAC4 gene rs1108519 locus polymorphism is related to hypertension, and the G allele may be a risk factor for the onset of essential hypertension.

OBJECTIVE：To study the effects of Modified liuwei dihuang decoction on kidney/bone injury of chronic kidney disease-mineral and bone disorder（CKD-MBD）model rats. METHODS ：The male SD rats were randomly divided into normal group（n＝10），high phosphorus group （n＝30），model group （n＝30），calcitriol group （positive control ，0.09 μg/kg，n＝30）， Modified liuwei dihuang decoction group （10 g/kg by crude drug ，n＝30）. CKD-MBD model was established by high phosphorus and adenine diet for 6 weeks. After modeling ，normal group and model group were given normal diet/high phosphorus diet and intragastric administration of water. Administration groups were fed with normal diet and given corresponding solution intragastrically（water as solvent ），0.1 mL/kg，once a day ，for consecutive 6 weeks. Blood sample of rats in the normal group were collected ，and they were sacrificed after the last administration. Blood sample of 10 rats in each other group were collected ， and they were sacrificed at 2，4 and 6 weeks after administration. The contents of blood urea nitrogen （BUN），serum creatinine （Scr），calcium，phosphorus，iPTH，FGF-23，RANKL and osteocalcin in serum were detected in each group. The bone mineral density（BMD）of femoral was measured ，the morphological changes of renal tissue and bone tissue were observed ，and the percentage of renal tubular injury and the score of renal interstitial fibrosis were calculated. RESULTS ：Compared with normal group，above indexes in high phosphorus group had no significant change at different time points （P＞0.05）. There was no abnormal change in renal/bone tissue. Compared with high phosphorus group at the same time point ，the contents of BUN ，Scr， phosphorus，iPTH，FGF-23，RANKL and osteocalcin in serum ，the percentage of renal tubular injury and the score of renal interstitial fibrosis in the model group were significantly increased ，while the contents of calcium in serum and the BMD of femoral were significantly decreased （P＜0.05 or P＜0.01）. The renal tissue showed diffuse fibrosis. The width of trabecular bone was increased and the number of osteoblasts was decreased. Compared with the model group at the same time point ，the contents of BUN（except for Modified liuwei dihuang decoction group after 2 weeks of administration ），Scr，serum phosphorus ，iPTH， FGF-23，RANKL and osteocalcin ，the percentage of renal tubular injury and the score of renal interstitial fibrosis in Modified liuwei dihuang decoction group and calcitriol group were decreased significantly at each time point ；serum calcium content and BMD（except for 2 weeks of administration ）were significantly increased （P＜0.05 or P＜0.01），and the pathological changes of renal/bone tissue were significantly improved ；there was no statistical significance in above indexes between Modified liuwei dihuang decoction group and calcitriol group （P＞0.05）. CONCLUSIONS ：Modified liuwei dihuang decoction can improve kidney/ bone injury of CKD-MBD model rats ，and improve BMD and regulate disorder of calcium and phosphorus metabolism.

Objective: To investigate the clinical characteristics and risk factors of congenital choledochal cysts (CCC). Methods: This retrospective study recruited 52 cases who were antenatally diagnosed with CCC and underwent surgical treatment after birth in Guangdong Women and Children Hospital from January 2013 to August 2018, with complete clinical data. According to the enlargement of cysts during pregnancy, they were divided into two groups: progressive group (≥15 mm, 22) and stable group (<15 mm, 30). Antenatal and postpartum ultrasound and MRI features of the two groups were analyzed. Clinical manifestations and biochemical examination results before and after operation were compared between the two groups. Other data, including amylase level in cyst fluid during operation, cholangiography findings, liver biopsy results, and post-operation follow-up, were also analyzed. Chi-square test, t (t') test, and Pearson correlations tests were performed for data analysis. Results: (1) The average age of the 52 patients at operation was 46(7-822) d. The cysts of all cases were first detected during 19-21 weeks of gestation. The maximum diameter of the cyst in the progressive group was larger than that in the stable group after 34 weeks of pregnancy [31-34 weeks: (31.1±8.4) vs (23.1±6.6) mm, t=3.911; >34 weeks: (36.1±6.8) vs (27.1±7.3) mm, t=4.557; pre-operation: (51.8±18.0) vs (34.0±15.6) mm, t=3.809; all P<0.01]. (2) In the progressive group, the cysts were irregular in shape and enlarged after birth. The common hepatic duct and intrahepatic bile duct were dilated and gradually distended after birth, while the distal end of the common bile duct was narrowed, thus to form a cone-like duct. Deposits could be seen inside the cysts after delivery. Irregular cysts were also presented in the stable group, and five of them had dilatation of common hepatic duct and intrahepatic bile duct after birth. However, no cone-like formation was seen, the distal end of the common bile duct was visible, and deposits in cysts were occasionally found. (3) Twenty-five patients underwent laparotomy, and seven of them showed increased amylase level in cyst fluid including four with 2-5 times above the upper limit of normal value (one in the progressive group and three in the stable group). The other three cases were all in the stable group and their amylase levels in cyst fluid were more than ten times of the upper limit. The level of direct bilirubin in the progressive group was higher than that in the stable group before the operation [18.40(2.50-113.30) vs 8.70(0.00-16.80) μmol/L, u=2.400, P<0.05]. (4) Among the 52 cases, patients with type Ⅰ, Ⅳ and Ⅴ cyst accounted for 71.1% (37/52), 26.9% (14/52) and 2.0% (1/52), respectively. All cases were followed up regularly six months to one year after the operation. Liver function and bilirubin became normal and the growth and development of the babies were similar to those of the same age. (5) Different degrees of liver fibrosis and inflammation were shown in 46(88.5%) cases and more severe in older babies among those who underwent surgery in the progressive group. The time at operation was not associated with the severity of liver fibrosis and inflammation in the stable group. Hepatic fibrosis and inflammation were more serious in the progressive group than in the stable group (fibrosis grading: χ²=14.260, P=0.006; inflammatory activity grading: χ²=9.904, P=0.019). Conclusions Larger diameter (≥30 mm) in the initial prenatal examination or a significant increase in cystic diameter (≥15 mm) during pregnancy are risk factors for early stenosis or occlusion in the distal end of common bile duct requiring close follow-up after birth. When jaundice or abnormal liver function occur and stool color becomes light, early surgical treatment (one to two months after birth, generally within three months) for CCC is recommended to rule out the anomalous union of the pancreaticobiliary duct and hepatic disorders, as well as the cystic biliary atresia.