abstract:
Liver metastasis is one of the most common complications of advanced lung cancer and an important fac- tor influencing patient prognosis and survival . Currently , there are limited effective treatment options for lung canc- er patients with liver metastasis , leading to short survival and poor prognosis . In-depth studies of the related molec- ular mechanisms are crucial for advancing clinical translation and optimizing therapeutic strategies . In recent years , more and more studies of the mechanisms of liver metastasis in lung cancer have performed , particularly in areas such as the roles of different proteins , cell-cell interactions , and changes in the tumor microenvironment. This re- view summarizes the current understanding of the basic process of lung cancer liver metastasis , regulatory proteins , signaling pathways , tumor microenvironment changes , and clinical treatment progress . E merging evidence high- lights the critical involvement of TGF-β/smad signaling pathway and integrin family proteins in promoting lung cancer liver metastasis . In the tumor microenvironment , various cell types including mononuclear phagocytes , fi- broblasts , and hepatocytes contribute to this metastatic process . Clinically , the combination of immunotherapy with chemotherapy , radiotherapy , and antiangiogenic therapy has shown potential to improve treatment outcomes . Fur- thermore , targeted therapy against specific pathways , proteins , and cells within the tumor microenvironment , as well as the integration of multiple treatment modalities , holds promise for becoming effective strategies in the future clinical management of lung cancer liver metastasis .

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To investigate the characteristics of temporal and spatial distribution of tuberculosis epidem- ics in Shache County , Kashgar Region , Xinjiang. Methods Information on the incidence of tuberculosis in Sacha County from 2019—2021 was collected and spatiotemporally analyzed by applying the circular distribution method , local spatial autocorrelation analysis , hot and cold spot analysis , directional distribution and spatial center of gravity methods . Results The total number of tuberculosis cases in Shache County in 2019—2021 was 8 , 345 , of which 52. 03% were male and 47. 97% were female , and the patients were predominantly 60 - 75 years old . The number of reported incidences of TB in Tagarqi Township , Shache Township , and Chajek Township ranked among the top three in the county . Spring and summer were the disease-prone seasons for TB , and mid-March to mid-July was the period of high disease incidence . Misha Township and Ishkuli Township are the “high and high ”gathering areas , while the “low and low ”gathering areas are mainly concentrated in Khoshrav Township and Karasu Township . The hotspots of TB incidence in Shache County were Tagarqi Township , Misha Township , and Ishkuli Township . During the study period , the center of gravity of TB incidence in shache county of kashgar area gradually shifted from the southwest to the northeast. Conclusion In Shache County , there is a certain degree of aggregation of tuberculosis outbreaks , with more men than women reporting illnesses , a larger proportion of older people , and a strong seasonal incidence of the disease , with Mixia Township and Ishikuli Township being the key areas of incidence . Relevant departments should continue to strengthen the disease surveillance of key populations and regions during the high incidence of tuberculosis , and take appropriate intervention measures to reduce the risk of tuberculosis transmis- sion .

abstract:
To explore the expression of prohibitin2 (PHB2) in breast cancer and its effect on the bio- logical behaviors of tumor cells . Methods Immunohistochemistry was used to detect the expression of PHB2 pro- tein in breast cancer tissues and its relationship with clinicopathologic features . Breast cancer stable transient cell lines were constructed with knockdown and overexpression of PHB2 , respectively. The effects of PHB2 on cell pro- liferation , migration and invasion ability were detected by clone formation assay , scratch assay and Transwell assay. Western blot (WB) was used to detect the effects of PHB2 on the expression of epithelial-mesenchymal transition (EMT) - related markers , including E-cadherin , N-cadherin , Snail family transcriptional repressor 1 (Snail) pro- tein , Vimentin , and Claudin-1 . The effect of PHB2 on tumorigenicity in vivo was detected by subcutaneous tumor formation assay in nude mice . Results The result of immunohistochemical showed that PHB2 was highly expressed in breast cancer and the expression of PHB2 was significantly positive correlated with tumor size , human epidermal growth factor receptor-2 (HER-2) status and proliferation index Ki-67 levels (P < 0. 05) . Clone formation assay , scratch assay and Transwell assay revealed that knockdown of significantly inhibited the proliferation , migration and invasion ability of breast cancer cells ( P < 0. 01) , while the overexpression of PHB2 significantly promoted cell proliferation , migration and invasion (P < 0. 01) . The result of subcutaneous tumor formation experiment in nude mice revealed a significant decrease in tumor volume and weight in knockdown PHB2 mice (P < 0. 000 1) , while PHB2 overexpression tumors significantly increased in volume and weight (P < 0. 001) . WB assay showed that the protein expression of epithelial marker E-cadherin increased , while the expressions of mesenchymal markers N-cad- herin , Snail and Vimentin decreased significantly after PHB2 knockdown with them in control cells (P < 0. 01) . The expression of Claudin-1 decreased , while the expressions of N-cadherin , Snail and Vimentin increased signifi- cantly in PHB2 overexpression cells (P < 0. 05) . Conclusion PHB2 is highly expressed in breast cancer and pro- motes multiple malignant biological behaviors in tumor cells , suggesting PHB2 may be a potential target for breast cancer diagnosis and treatment.

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To investigate the expression of Keratin 14 (KRT14) in Basal-like Breast Cancer (BLBC) and its biological functions and mechanisms . Methods The expression levels of KRT14 mRNA in BLBC and para- cancer breast tissues were analyzed using The Cancer Genome Atlas ( TCGA) database . qPCR , Western blot (WB) , and immunohistochemistry were employed to detect KRT14 expression in BLBC and adjacent normal tis- sues , and its correlation with clinicopathological features was analyzed . KRT14 overexpression and knockdown were performed in breast cancer cells , and cell scratch and transwell assays were performed to evaluate changes in migra- tion and invasion abilities . To investigate the expression of proteins related to the Wnt/β-catenin signaling pathway , including β-catenin ( catenin Beta 1) , Wnt1 ( wingless-type MMTV integration site family , member 1) , MMP7 ( matrix metallopeptidase 7) , and c-Myc (cellular myelocytomatosis viral oncogene homolog) , as well as the cellu- lar localization of β-catenin , WB and immunofluorescence (IF) techniques were employed . Additionally , a Wnt/ β-catenin signaling pathway inhibitor was used to verify the mechanism of action of KRT14 . Results The expres- sion of KRT14 was significantly higher in BLBC tissues compared to normal tissues (P < 0. 05) , and was associated with higher T stage , and histological grade( P < 0. 05) . The overexpression of KRT14 significantly enhanced the migration and invasion abilities of breast cancer cells , while the knockdown of KRT14 significantly reduced those a- bilities (P < 0. 01) . The overexpression of KRT14 can increase the expression levels of Wnt/β-catenin pathway-re- lated proteins β-catenin , Wnt1 , MMP7 , and c-Myc , thereby activating the Wnt/β-catenin pathway . Moreover , the inhibition of this pathway can eliminate the effects of KRT14 on cell migration and invasion . Conclusion The high expression of KRT14 in BLBC may promote the migration and invasion of breast cancer cells through the Wnt/β- catenin signaling pathway .

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To investigate the regulatory role of ABCG5 in head and neck squamous cell carcinoma. Methods Immunohistochemical staining was used to detect the expression of ABCG5 in 105 patients with hypopha- ryngeal carcinoma , and the relationship between the expression and the long-term survival time of the patients was analyzed . Immunowestern blotting and flow cytometry were used to detect the expression of ABCG5 in head and neck squamous cell carcinoma cell lines . Flow sorting was employed to investigate the differences in proliferation and metastasis , spheroidization ability , and expression of stem cell molecules between ABCG5 positive and negative cells . si RNA interference was used to further demonstrate whether ABCG5 affects the malignant phenotype . Re- sults ABCG5 was associated with poor prognosis in patients with hypopharyngeal carcinoma. ABCG5 was enriched in stem cells of head and neck squamous cell carcinoma. ABCG5-positive cells had stronger proliferation ( P < 0. 001), metastasis (P < 0. 01), and spheroid formation abilities (P < 0. 001) than negative cells . The expression levels of some stem cell molecules (SOX2 、NANOG、SOX9 、OCT4 、CD44) in ABCG5-positive cells were higher than those in ABCG5-negative cells . The expression of epithelial cell protein E-cadherin was lower in ABCG5-positive cells than that in ABCG5-negative cells , and the expression of interstitial cell proteins N-cadherin , Slug , Snail 1 , and Vimentin were higher (P < 0. 01) . Moreover , interfering with ABCG5 expression significantly inhibited tumor cell spheroid formation (P < 0. 001), as well as the expression of proteins related to cancer stem cells and epitheli- al-mesenchymal transition . Conclusion ABCG5 has a potential biological role in maintaining the function of head and neck squamous cell carcinoma stem cells .

abstract:
To investigate the potential of bortezomib in inhibiting the malignant biological behaviors and liver metastasis of pancreatic cancer cells . Methods KPC and Panc02 cell lines were used as a couple of working cells in this study . The CCK-8 assay was used to assess the effect of bortezomib on cell viability in murine pancreatic cancer cells , and the half inhibitory concentration ( IC50 ) values were calculated . The EdU assay was performed to evaluate the impact of bortezomib on the proliferation of these two cell lines . Apoptosis assays were conducted to investigate the pro-apoptotic effects of bortezomib on pancreatic cancer cells . Colony formation , scratch wound healing , and Transwell assays were used to examine the effects of bortezomib on the proliferation , migration , and invasion of pancreatic cancer cells . QRT-PCR and Western blot were employed to assess the impact of bortezomib on the expression of epithelial-mesenchymal transition ( EMT) related markers , including Cdh1 , Cdh2 , Vim and Snail gene and E-cadherin protein , N-cadherin protein , Vimentin protein and Snail protein . An in vivo spleen-to-liver metastasis model was established to evaluate the therapeutic value of bortezomib in inhibiting pancreatic cancer liver metastasis . Results The CCK-8 assay showed that bortezomib significantly reduced the via- bility of KPC and Panc02 cells (P < 0. 000 1) , with IC50 values of approximately 118. 70 nmol/L and 34. 16 nmol/ L , respectively . The EdU assay showed that bortezomib markedly inhibited the proliferative capacity of pancreatic cancer cells (P < 0. 01) . Apoptosis assays showed that bortezomib promoted both early and late apoptosis in pan- creatic cancer cells (P < 0. 05) . Colony formation , scratch wound healing , and Transwell assays showed that bort- ezomib effectively suppressed colony formation ( P < 0. 01) , migration , and invasion ( P < 0. 01) of pancreatic cancer cells . qRT-PCR and Western blot analyses showed that bortezomib altered the expression of EMT-related markers at both the mRNA ( the expression level of Cdh1 gene increased , while the expression levels of Cdh2 , Vim and Snail genes decreased . ) ( P < 0. 000 1) and protein ( the expression level of E-cadherin protein increased , while the expression levels of N-cadherin protein , Vimentin protein and Snail protein decreased) levels in pancreat- ic cancer cells . The in vivo spleen-to-liver metastasis model demonstrated that bortezomib significantly inhibited pancreatic cancer liver metastasis ( P < 0. 01) . Conclusion Bortezomib can inhibit the malignant biological be- haviors of pancreatic cancer cells , suggesting it might be a potential anti-cancer drug in pancreatic cancer.

abstract:
To develop and to validate a combined model integrating 18 F-FDG PET/CT radiomics with clinical features to distinguish between lymphoma and lymphoid inflammatory hyperplasia. Methods A retrospec- tive study was conducted on a cohort of 232 patients diagnosed with lymphoma or lymphoid inflammatory hyperplasi- a. Comparative analyses of clinical and traditional imaging indicators were performed to identify inter-group differ- ences . The clinical features were delineated and extracted using medical software including 3DSlicer and Lifex . Se- lection of the features was performed to construct a PET/CT-based radiomics Logistic model , with a combined mod- el integrating PET/CT with clinical features then used to evaluate the discriminative efficacy of these models . Re- sults Analysis of inter-group differences indicated that age , CTmean , and metabolic tumor volume (MTV) were ef- fective for differentiating between lymphoma and lymphoid inflammatory hyperplasia ( P < 0. 05 ) . The PET/CT- based radiomics Logistic model differentiated between lymphoma and lymphoid inflammatory hyperplasia , with an area under curve (AUC) of 0. 924 (95% CI: 0. 884 - 0. 960) and 0. 863 (95% CI: 0. 774 - 0. 939) in the training and testing cohorts , respectively . The integrated Logistic model that combined PET/CT-based radiomics with clinical features to distinguish between lymphoma and lymphoid inflammatory hyperplasia achieved an AUC of 0. 933 (95% CI: 0. 889 - 0. 969) in the training cohort and 0. 884 (95% CI: 0. 792 - 0. 964) in the testing co- hort . Decision curve analysis (DCA) demonstrated that the integrated model provided the greatest clinical net ben- efit. Conclusion The hybrid model integrating 18 F-FDG PET/CT radiomics with clinical features shows robust di- agnostic efficacy to distinguish between lymphoma and lymphoid inflammatory hyperplasia.

abstract:
To investigate the expression of HSPA8 in breast cancer and its effect on tumor biological behaviors . Methods Bioinformatics analysis and immunohistochemistry assays were used to detect the expression of HSPA8 in breast cancer and adjacent non-tumor breast tissues , and the relationship between its expression and clinicopathological characteristics of breast cancer patients was analyzed . Correlation between HSPA8 expression and prognosis of breast cancer patients was analyzed by Kaplan-Meier Plotter database . HSPA8 knockdown and overexpression breast cancer stabilized cells were constructed , respectively. CCK-8 , clone formation , Transwell , cell scratch , Western blot and immunofluorescence assay were used to detect the effects of HSPA8 on the prolifera- tion , invasion and migration of breast cancer cells , and its effect on epithelial-mesenchymal transition (EMT) . Results Bioinformatics analysis and immunohistochemistry assay revealed that the expression of HSPA8 in breast cancer tissues was higher than that in adjacent non-tumour tissues (P < 0. 05) , and its expression level of the pro- tein was significantly and positively correlated with the tumor size , histological grade , lymph node metastasis and Ki-67 proliferation index (P < 0. 05) . The Kaplan-Meier survival curve showed that high expression of HSPA8 was significantly associated with poor prognosis in breast cancer patients (P < 0. 05) . CCK-8 , clone formation , tran- swell , cell scratch , Western blot and immunofluorescence assay showed that knockdown of HSPA8 expression could significantly inhibit the proliferation , invasion , migration function and EMT of breast cancer cells ( P < 0. 05 ) , while overexpression of HSPA8 could significantly promote the proliferation , invasion , migration function and EMT of breast cancer cells ( P < 0. 05) . Conclusion HSPA8 is highly expressed in breast cancer tissues , which is closely related to disease progression and the malignant phenotype of breast cancer , suggesting that HSPA8 may be a potential biological target for breast cancer treatment.

abstract:
To explore the role of semaphoring 6d ( SEMA6D) in the malignant progression of triple- negative breast cancer (TNBC) . Methods Bioinformatics and Immunohistochemistry (IHC) were used to analyze the expression level of SEMA6D in TNBC and paracancer non-tumor tissues and its relationship with patients ′clini- copathological features . MDA-MB-231 cell line stably knocking down the expression of SEMA6D was constructed , and the effects of SEMA6D on migration and invasion of TNBC cells were investigated by Wound-healing assays and Transwell assays . cBioPortal and GEPIA2 databases were used to screen out the gene negatively associated with it , namely aurora kinase A (AURKA) . Bioinformatics and IHC were used to analyze the expression level of AURKA in TNBC and paracancer non-tumor tissues and its relationship with patients ’clinicopathological features . Western blot assay was used to analyze the expression of AURKA and the effect of epithelial-mesenchymal transition (EMT) makers Claudin-1 , N-cadherin and Vimentin after knocking down SEMA6D . Results Bioinformatics analysis and IHC results showed that the expression of SEMA6D in TNBC tissues was significantly lower than that in paracancer non-tumor tissues (both P < 0. 05) . The expression of AURKA in TNBC tissues was significantly higher than that in paracancer non-tumor tissues (both P < 0. 05) , SEMA6D and AURKA were significantly negatively correlated in TNBC (P < 0. 01) . Both low expression of SEMA6D and high expression of AURKA were positively correlated with tumor size , tumor histological grade , clinical stage and lymph node metastasis in TNBC patients (all P < 0. 05) . The knockdown of SEMA6D significantly promoted the migration and invasion of ability TNBC cells ( both P < 0. 01) . Western blot results showed that the knockdown of SEMA6D upregulated AURKA expression , promoted the expression of N-cadherin and Vimentin , and inhibited the expression of Claudin-1 in tumor cells . Conclusion Down-regulation of SEMA6D expression in TNBC may be involved in the malignant progression of TNBC through up-regulation of AURKA expression and promotion of EMT.

abstract:
To construct CSF1R + / - mice and to analyze their genotypes , so as to provide animal model basis for disease pathological mechanism and drug target. Methods A linearized targeting vector was designed ac- cording to Cre/Loxp system . A Loxp site was inserted upstream of the 5th exon of the CSF1R gene , and a neomycin resistance box with Loxp sites on both sides was inserted downstream of the 5th exon . The linearized targeting vector was electroporated into embryonic stem cells . The correctly targeted embryonic stem cells were injected into the blastocysts of C57BL/6J mice to obtain chimeric mice , which were bred with Zp3-Cre mice . The newborn mice were numbered 9 - 14 days after birth and their tails were cut. The DNA of the mice was extracted , and the geno- type of the mice was identified by polymerase chain reaction and agarose gel electrophoresis . The expression of CSF1R in mouse macrophages was detected by flow cytometry . The expression of CSF1R in mouse tissues was de- tected by Western blot. Results The results of agarose gel electrophoresis showed that 453 bp bands were ampli- fied in wild type mice , and 453 bp and 650 bp bands were amplified in heterozygous mice . The results of flow cy- tometry showed that the expression of CSF1R in peritoneal macrophages and bone marrow-derived macrophages of CSF1R heterozygous mice was lower than that of WT group ( P < 0. 05) . The results of Western blot showed that the expression of CSF1R in spleen , kidney and brain tissue of CSF1R heterozygous group was lower than that of WT group (P < 0. 05) . Conclusion CSF1R + / - mice are successfully constructed , reproduced and identified , which provides an animal model basis for further revealing the potential mechanism of CSF1R in immune regulation .

abstract:
To investigate the effect of phosphatase and tensin homolog (PTEN) overexpression on au- tophagy in mouse primary cardiac fibroblasts . Methods Neonatal mice ( 1 - 3 days old) were purchased and sub- jected to cardiac tissue harvesting. Cardiac fibroblasts were isolated through enzymatic digestion and cultured . After cellular adhesion , rapamycin (Rapa)-treated cardiac fibroblasts were used to establish an autophagy model . Fol- lowing successful model construction , cells were transfected with either PTEN-overexpressing plasmid (PTEN over- expression group) or empty vector (control group) , followed by 24 - 48 h incubation . The molecular expressions of PTEN , autophagy-related proteins ( microtubule-associated protein light chain 3 [ LC3 Ⅱ/ Ⅰ ] , Beclin1) , and fi- brotic marker collagen Ⅰ were detected by Western blot and RT-qPCR. Autophagic flux was assessed using mCher- ry-GFP-LC3 fluorescence staining to evaluate changes in autophagic activity. Finally , transmission electron micros- copy (TEM) was employed to observe autophagosome formation in cardiac fibroblasts . Results In the Rapa-in- duced cardiac fibroblast autophagy model , compared with the control group , the protein and mRNA levels of PTEN significantly decreased (P < 0. 05) , while the expression of autophagy-related proteins (LC3 Ⅱ/ Ⅰ , Beclin1) and fibrotic marker Collagen Ⅰ was upregulated at both protein and mRNA levels (P < 0. 05) . Additionally , in PTEN- overexpressing cardiac fibroblasts , the expression levels of LC3 Ⅱ/ Ⅰ , Beclin1 , and Collagen Ⅰ were markedly re- duced compared to the empty vector group (P < 0. 05) . mCherry-GFP-LC3 fluorescence staining demonstrated that autophagic activity was significantly attenuated in the PTEN overexpression group versus the empty vector group (P < 0. 05) . TEM further revealed a decreased number of autophagosomes in PTEN-overexpressing cardiac fibroblasts (P < 0. 05) . Conclusion The overexpression of PTEN significantly inhibits autophagy in cardiac fibroblasts , sug- gesting that PTEN may be a key gene involved in regulating autophagy in cardiac fibroblasts .

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To investigate the expression level of LONP1 in hepatocellular carcinoma ( HCC) and its impact on the occurrence and progression of HCC . Methods The expression of LONP1 in human liver cancer tis- sues was verified by real-time quantitative PCR (qPCR) and Western blot. LONP1 stable knockdown Hep3B and HCCLM3 cell lines were established , and the effects of LONP1 on cell proliferation were explored through Cell Counting Kit-8 (CCK-8) , EdU incorporation assays , and colony formation assays . The effects of LONP1 on cell migration were assessed using scratch and Transwell migration assays . A Cre-Loxp system was employed to generate LONP1 conditional knockout mice , and transcriptomic sequencing of liver tissues was performed to explore the im- pact of LONP1 deficiency on liver cells . The effects of LONP1 on apoptosis in hepatocellular carcinoma cell lines were explored using Tunel staining and flow cytometry with Annexin V-FITC/PI . Results Western blot and qPCR experiments confirmed the high expression of LONP1 in human liver cancer tissues . Colony formation assays re- vealed that the number of cell clones in LONP1 knockdown groups was significantly reduced compared to the control (P < 0. 01) . CCK-8 and EdU assays demonstrated that LONP1 knockdown cells had a significantly lower prolifera- tion rate than control cells (P < 0. 01) . Scratch and migration assays showed that LONP1 knockdown liver cancer cells exhibited impaired migration compared to controls (P < 0. 01) . Transcriptomic analysis of liver tissues from LONP1 conditional knockout mice indicated that LONP1 might affect apoptosis pathways in liver cells . Tunel stai- ning and Annexin V-FITC/PI flow cytometry showed that LONP1 knockdown increased apoptosis in hepatocellular carcinoma cells . Conclusion LONP1 is highly expressed in liver cancer tissues . The knockdown of LONP1 in liv- er cancer cell lines promotes cell apoptosis and inhibits cell proliferation and migration .

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To explore the role and mechanism of plasmalemma vesicle-associated protein (PLVAP) in the progression of hepatocellular carcinoma (HCC) . Methods Bioinformatic analysis , quantitative real-time PCR , Western blot and immunohistochemistry were used to analyze the expression level of PLVAP in HCC and paracan- cerous tissues , and its correlation with clinicopathological characteristics . Stable HCC cell lines with knockdown and overexpression of PLVAP were constructed , then cell proliferation , migration and invasion of HCC cells were examined by CCK-8 , foci formation assays , wound-healing assays and Transwell assays . Western blot was used to detect the protein levels of phosphatidylinositol 3-kinase PI3K , phosphorylated phosphatidylinositol 3-kinase p- PI3K , protein kinase B AKT and phosphorylated protein kinase B p-AKT in the PI3K/AKT pathway after PLVAP knockdown and overexpression . Cell proliferation , migration and invasion were also examined in PLVAP-overex- pressed cells after treatment of LY294002 , an inhibitor of the PI3K/AKT pathway . Results The expression of PLVAP was significantly higher in HCC tissues than that in adjacent non-tumor tissues (P < 0. 05) , and was posi- tive correlated with tumor Stage , T stage , M stage , and microvascular invasion (P < 0. 05) . Knockdown of PLVAP significantly reduced the proliferation , migration and invasion of HCC cells (P < 0. 001) , while overexpression of PLVAP significantly increased the proliferation , migration and invasion of HCC cells (P < 0. 01) . Western blot a- nalysis revealed that knockdown of PLVAP decreased the protein expression levels of p-PI3K and p-AKT , overex- pression of PLVAP increased the protein expression levels of p-PI3K and p-AKT , whereas the PI3K/AKT inhibitor LY294002 eliminated the effects of PLVAP on cell proliferation , migration and invasion (P < 0. 01) . Conclusion PLVAP is highly expressed in HCC and may promote HCC progression by activating the PI3K/AKT signaling pathway .

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To investigate the regulatory mechanism of nucleotide excision repair (NER) in hepatocel- lular carcinoma (HCC) . Methods Based on the expression levels of genes in the NER pathway , we performed molecular typing of HCC using the TCGA database . HCC cell lines were constructed through the knockdown of RNA binding motif protein 39 (RBM39) using siRNA . HCC cell lines were constructed through the overexpression of RBM39 using RBM39 plasmid . Cells were treated with Indisulam , a reagent that induces RBM39 protein degra- dation . Western blot and real-time fluorescence quantitative PCR were used to detect the expression levels and changes of mRNA and protein of RBM39 and excision repair cross complementation group 1 (ERCC1) ; flow cytom- etry was used to detect NER efficiency; CCK-8 assay was used to detect cell viability . Results HCC patients were categorized into three types—C1 , C2 , and C3—based on NER activity , with the C3 subtype showing the highest NER activity (P < 0. 0001) . In the groups transfected with RBM39 siRNA or treated with Indisulam , the NER re- pair efficiency decreased compared to the control group (P < 0. 01) , the cell survival rate decreased (P < 0. 01) , and both the mRNA and protein expression of ERCC1 were reduced (P < 0. 01) . In contrast , in the RBM39 over- expression group , the mRNA and protein expression of ERCC1 were enhanced compared to the control group (P < 0. 01) . Conclusion RBM39 may influence NER repair efficiency by regulating ERCC1 expression in HCC .

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To investigate the correlations among clinicopathological features , histopathological growth patterns and prognosis of extrapulmonary multiple metastatic (stage ⅣB) pulmonary adenocarcinoma with epider- mal growth factor receptor (EGFR) mutations . Methods A total of 488 eligible patients with adenocarcinoma of stage ⅣB from the First Affiliated Hospital of Anhui Medical University and the First Affiliated Hospital of Universi- ty of Science and Technology of China were collected . Clinicopathological data , EGFR gene mutation subtypes , metastatic sites , histopathological growth patterns and survival information were collected . The chi-square test ( χ2 test ) and Fisher exact probability method were used to detect the correlation between the metastasis status and va- rious clinical characteristics; the Kaplan-Meier method was used to conduct survival analysis on the median Pro- gression-Free Survival (PFS) under different clinical characteristics . Cox univariate and multivariate regression an- alyses were conducted to evaluate the impact of various clinical characteristics on prognosis . Results The metastat- ic patterns of stage Ⅳ B pulmonary adenocarcinoma with EGFR mutations was correlated with histopathological growth patterns (P < 0. 05) . In the group with multiple metastases in a single organ , the proportion of micropapil- lary type in the group with multiple metastases in a single organ was higher than that in the group with multiple-or- gan metastases (51 . 1% vs 41 . 1% ) , while the proportion of solid type in the group with multiple-organ metastases was higher than that in the group with multiple metastases in a single organ (23 . 8% vs 14. 2% ) . Multiple brain or multiple bone metastases were correlated with histopathological growth patterns and tumor differentiation degree . Compared with the multiple bone metastases group , the proportion of acinar type decreases in the multiple brain metastasis group , while the proportion of micropapillary type increased . Moreover , the proportion of poorly differen- tiated tumors increased significantly (P < 0. 05) . Compared with multiple bone metastases , the proportion of poorly differentiated tumors significantly increases in the group with multiple brain metastases . The median progression- free survival (PFS) of patients with a predominant solid growth pattern was shorter than that of patients with other growth patterns ( 12. 7 months vs 17. 8 months , P < 0. 05) . The PFS of patients in the poorly differentiated group was worse than that in the moderately differentiated group (15 . 6 months vs 17. 8 months , P < 0. 05) . There were significant differences in PFS among patients with common sensitive mutations and rare mutations EGFR ( 17. 3 months vs 10. 2 months , P < 0. 005) . Cox proportional hazards regression model suggested that solid growth pat- tern , poor differentiation and rare single gene mutation were adverse prognostic factors . Conclusion In stage ⅣB pulmonary adenocarcinoma patients with EGFR mutations , both the metastatic patterns and metastatic sites are sig- nificantly correlated with the histopathological growth patterns of tumors . Moreover , the EGFR mutation subtypes as well as the histopathological growth patterns and differentiation degree of tumors significantly affect the prognosis of patients .

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To explore the correlation between the clinical , pathological , genetic features , prognosis , and tumor lymph node metastasis in patients with stage Ⅰ A - Ⅲ B lung invasive non-mucinous adenocarcinoma (INMA) . Methods A retrospective analysis was conducted on 67 eligible patients with INMA . Clinical data , his- topathological assessments , and genetic testing were collected . Disease progression-free survival (PFS) was the pri- mary endpoint through follow-up . The chi-square test or Fisher exact test was used to analyse the correlation be- tween tumour lymph node metastasis and clinicopathological and genetic characteristics . The Cox proportional haz- ards regression model and Kaplan - Meier method were used to analyse the impact of tumour lymph node metastasis on prognosis . Results A total of 67 patients were included , aged 46 - 77 years , with a median age of 61 years . Age , gender , and smoking history were not significantly associated with tumor lymph node metastasis . Larger tumor diameter , tumor progression , and receiving postoperative adjuvant treatment were associated with tumour lymph node metastasis ( P < 0. 05) . Poorer differentiated tumors according to International Association for the Study of Lung Cancer (IASLC) grading system was more likely to have lymph node metastasis (P = 0. 043) . There was no significant difference in the types of driver gene mutations and lymph node metastasis . However , EGFR mutations were more common in patients without lymph node metastasis , while co-mutations were more common in patients with lymph node metastasis . Lymph node metastasis was significantly associated with PFS . Patients without lymph node metastasis had a significantly better PFS compared to those with lymph node metastasis (P = 0. 002) . Under different treatment conditions , patients without lymph node metastasis exhibited a significant advantage in PFS when untreated . While treatment showed a trend toward improved PFS , the difference did not reach statistical signifi- cance . Additionally , no significant differences in PFS were observed between patients with or without lymph node metastasis following chemotherapy or targeted therapy . Conclusion Lymph node metastasis in INMA patients is re- lated to tumor size , progression status , and gene co-mutations , and is a key prognostic indicator affecting PFS .

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To investigate the effect and mechanism of methyltransferase-like3 (METTL3) , which func- tions as m6 A modification methyltransferase , on the cellular proliferation , clone formation and migration of non- small cell lung cancer (NSCLC) cells . Methods The METTL3 level and patient overall survival were analyzed by bioinformatics . In the transfected A549 cells , the expression of METTL3 was detected by RT-PCR and Western blot assays , the m6 A level was detected by m6 A RNA Methylation Assay kit , the cellular viability and growth were de- tected by CCK8 assay , the apoptosis was detected by immunofluorescent assay , the colony growth was detected by clone formation assay , the cell migration growth was detected by scratch . Results Bioinformatics showed that METTL3 was highly expressed in NSCLC and negatively correlated with patient overall survival . The RT-PCR and Western blot data showed that the METTL3 level was higher in NSCLC cells . Moreover , higher METTL3 signifi- cantly promoted m6 A level , cell proliferation , colony formation , migration , inhibited cell apoptosis , increased the mRNA and protein expressions of EMT-related marker Vimentin but inhibited apoptosis and decreased the mRNA and protein expressions of EMT-related marker E-cadherin in the A549 cells . Furthermore , the contrasting results were obtained in the A549 cells with the knockdown of METTL3 . Conclusion The over-expressed METTL3 in- creases the m6 A levels in NSCLC cells and promotes cellular proliferation , colony formation , and migration growth , thereby laying a theoretical foundation for future research on early diagnosis and targeted drug development for NSCLC by targeting METTL3 .

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To illuminate the distribution of brucellosis patients and the epidemic typologies as well as the genetic attributes of brucellosis in Bozhou City , Anhui Province , thereby furnishing a substantive foundation for formulating efficacious prevention and control strategies for this disease within the region . Methods The rose ben- gal plate agglutination test (RBPT) and the tube agglutination test (TAT) were conducted on a total of 698 blood samples that had been collected . Epidemiological data of the tested subjects were meticulously collected , followed by statistical analyses of the obtained results . The genomic DNA of positive bacterial strains was cultured and ex- tracted . Molecular identification and typing of the isolated strains were executed through 16S rRNA sequencing. Se- quence alignment was conducted employing Clustal W and MEGA 7 , with comparisons made against the outcomes of AMOS-PCR and BCSP31-PCR. Results A total of 66 positive samples were detected through serological assays , with a positive rate of 9. 46% . The demographic cohort demonstrating the highest detection rate primarily comprised individuals engaged in live sheep slaughtering. The 16S rRNA gene sequencing on ten positive strains disclosed close phylogenetic affinities with Brucella melitensis . Moreover , the phylogenetic tree analysis indicated that these strains coalesced within the same branch , the findings were in alignment with the results obtained from BCSP31 - PCR and AMOS-PCR assays . Conclusion Brucella melitensis assumes a predominant position in the transmission dynamics within this area , identifying individuals involved in sheep breeding , slaughtering , vending , and related occupations as high-risk groups . The outcomes of this study offer molecular biological substantiation for the distribu- tion of brucellosis patients in this region , contribute to genotyping endeavors and tracing studies associated with the pathogen , and concurrently verify the efficacy of 16S rRNA molecular tracing.

abstract:
To characterize MMP12 as a pan-cancer marker and assess its screening value as a tumor serum marker. Methods Bioinformatics tools such as GEPIA2 , GSCA , cBioPortal , and GeneMANIA were used to analyze the pan-cancer features of MMP12 in TCGA datasets , including encompassing differential gene expression analysis , prognostic analysis , DNA methylation analysis , gene structural variation analysis and immune microenvi- ronment analysis . Furthermore , serum samples we collected from patients with lung adenocarcinoma , breast inva- sive carcinoma , esophageal squamous cell carcinoma , stomach adenocarcinoma , liver hepatocellular carcinoma , and healthy individuals . ELISA was used to detect MMP12 expression in serum , and the screening performance was evaluated using the area under the ROC curve . Additionally , we followed up 28 ESCC patients and compared serum MMP12 levels between 19 patients with disease progression and 9 patients with stable disease . Results The pan- cancer feature analysis revealed a significant negative correlation between MMP12 mRNA expression and its promot-er DNA methylation (P < 0. 05) , as well as a positive correlation with gene copy number variations (P < 0. 05) . MMP12 mRNA expression was up-regulated in 14 cancer tissues compared to normal tissues next to cancer (P < 0. 05) and was associated with poor prognosis of cancer patients (P < 0. 05) . Immunocorrelation analysis showed that MMP12 was significantly associated with immunity , infiltration of stromal cells , tumor mutational burden (TMB) and microsatellite instability (MSI) (P < 0. 05) . ROC curve analysis indicated that MMP12 could serve as a potential biomarker for screening lung adenocarcinoma , breast invasive carcinoma , esophageal squamous cell car- cinoma , stomach adenocarcinoma , and liver hepatocellular carcinoma. In a 30-month follow-up study of esophageal squamous cell carcinoma patients , the expression of MMP12 was higher in the disease progression group than that in the stable group . Conclusion MMP12 serves as a potential prognostic and screening marker of pan-cancer.

abstract:
To investigate the mechanism of action of curcumol on mitochondrial autophagy in hepatic stellate cells and its molecular mechanism against liver fibrosis. Methods Hepatic stellate cells were divided into blank group , model group ( lipopolysaccharide 5 mg/L) , and low , medium and high curcumol group ( 12. 5 , 25 and 50mg/L) ; Thiazolyland (MTT) was used to detect the effects of curcumol on the viability of hepatic stellate cells; flow cytometry was used to detect the effects of curcumol on apoptosis of hepatic stellate cells; 5 , 5 ′,6 , 6 ′- Tetrachloro-1 , 1 ′,3 , 3 ′-tetraethylimidacarbocyanine iodide ( JC-1) was used to detect the mitochondrial membrane potential; effects of curcumol on mitochondrial morphology and autophagosome were detected by transmission elec- tron microscopy; effect of curcumol on mitochondrial localisation were detected by fluorescent probe; Immunoblot- ting assay was performed to detect the effects of curcumin on PTEN-induced putative kinase 1 (PINK1) , Parkin- son ’s disease protein (Parkin) , microtubule-associated protein light chain 3 (LC3) , autophagy-associated protein (Beclin1) , mitochondrial inner membrane translocase 23 ( Timm23) , mitochondrial outer membrane translocase 20 ( Tomm20 ) , Bcl-2 associated X protein ( Bax) , B lymphocytoma-2 ( Bcl2 ) , cleaved-cysteine protease 3 (Caspase3) , α-smooth muscle actin ( ɑ-SMA) , collagen type Ⅰ (Collagen Ⅰ ) , and collagen type Ⅲ (Collagen Ⅲ) protein expression . Results Compared with the blank control group , cell proliferation rate , Caspase3 , Bcl2 , LC3 Ⅱ , Beclin1 , PINK1 , Parkin , ɑ-SMA , Collagen Ⅰ , CollagenⅢ proteins significantly increased in the model group (P < 0. 01) , co-localisation of mitochondria and lysosomes increased , and the number of mitochondrial auto- phagosome significantly increased (P < 0. 01) , while Timm23 and Tomm20 proteins , mitochondrial membrane po- tential decreased significantly (P < 0. 01) , apoptosis rate decreased , and Bax protein expression decreased . Com- pared with the model group , after curcumol intervention , cell proliferation rate , Bcl2 , Timm23 , Tomm20 , ɑ-SMA , Collagen Ⅰ and CollagenⅢ protein expression significantly decreased in the curcumol low - , medium - and high- concentration groups (P < 0. 01) , and the mitochondrial membrane potential significantly decreased (P < 0. 01) , whereas apoptosis rate , Caspase3 , Bax , LC3 Ⅱ , Beclin1 , PINK1 and Parkin proteins significantly increased (P < 0. 05) , the co-localisation of mitochondria and lysosomes increased , and the number of mitochondrial autophago- some significantly increased (P < 0. 01) . Conclusion Curcumol exerts ameliorative effects on hepatic fibrosis by modulating mitochondrial hyperautophagy mediated by the PINK1 /Parkin signaling pathway , and promoting hepatic stellate cell apoptosis .

abstract:
To investigate the biological functions and regulatory mechanism of hexokinase ( HK2) in driving tumor initiation and development of prostate cancer. Methods Immunohistochemical staining was employed to assess HK2 expression in benign prostate tissue and prostate cancer tissue . Correlation analyses were performed to elucidate the association of HK2 expression and clinical factors . Western blot analyses were employed to assess HK2 expression in prostate cancer cell lines . Cellular viability was assessed using the MTT assay . In vivo tumori- genesis ability of HK2 was evaluated using xenograft animal models . Glucose flux tracing was conducted through i- sotopic analyses in prostate cancer cells . Results HK2 expression was elevated in prostate cancer tissues compared to benign prostate tissues (P < 0. 05) . HK2 expression was positively correlated with Gleason Score , prostate-spe- cific antigen (PSA) levels and the presence of metastases ( P < 0. 05) . HK2 enhanced cell proliferation in vitro and tumorigenesis in vivo . HK2 augmented metabolic activity of glycolysis pathway in prostate cancer cells . Con- clusion HK2 facilitates tumorigenesis in prostate cancer by upregulating glycolytic activity .

abstract:
To prepare silk protein (SF) membranes reinforced by aragonite sheets (AP) and to inves- tigate their potential as novel barrier membranes for bone tissue regeneration by testing their tensile properties , bio- compatibility , and their effect on osteogenic differentiation . Methods AP was extracted from natural abalone shell (AS) by oxidation method , and AP-SF composite and pure SF membranes with different component ratios were pre- pared by solution casting and evaporative self-assembly techniques , and their tensile properties were tested using a universal mechanical machine under wet state conditions . The tensile properties were tested using a universal me- chanical machine under wet conditions . The microstructure of the AP-SF membrane with the highest tensile strength was observed using scanning electron microscopy (SEM) and characterized using Fourier transform infrared spec- troscopy (FTIR) and X-ray diffraction (XRD) . Observation of the adhesion of rat bone marrow mesenchymal stem cells (rBMSCs) on the surface by scanning electron microscopy , and exploration of the biocompatibility of AP-SF membrane by CCK-8 and live cell staining. Alkaline phosphatase staining (ALP) and alizarin red staining (ARS) were used to detect the osteogenic differentiation of rBMSCs in each group . Results AP was successfully extracted from AS , and the membrane prepared from a mixed solution of SF and AP with a solid content of 10 ∶9 had the strongest wet tensile performance , reaching 8. 46 MPa. The CCK-8 and live dead cell staining test results of the AP-SF membrane showed no significant difference compared to the blank group and SF group , indicating that both membranes had good biocompatibility. The ALP test and ARS results indicated that AP-SF membrane had a more significant ability to promote osteogenic differentiation of rBMSCs compared to SFmembrane . Conclusion The prepared AP-SF membrane has both good mechanical and biological properties , and has certain clinical application potential .

abstract:
To establish a method for measuring major organic acids in the tricarboxylic acid cycle using a high-performance liquid chromatography-tandem mass spectrometry ( HPLC-MS/MS) system , and to investigate the changes in six tricarboxylic acid cycle organic acids ( fumaric acid , malic acid , succinic acid , α-ketoglutaric acid , cis-aconitic acid , and citric acid) in the serum , liver , and placenta of mice exposed to di(2-ethylhexyl) phthalate (DEHP) during pregnancy . Methods The serum , liver and placental samples from pregnant mice were processed and eluted through a Waters ACQUITY UPLC BEH Amide Column ( 130 ?，1 . 7 μm , 2. 1 mm × 150 mm ) using a gradient elution program . Mobile phase A comprised an aqueous solution of 10 mmol/L ammonium acetate and 5 μmol/L methanephosphonic acid , while mobile phase B consisted of a 90% acetonitrile aqueous solu- tion containing 10 mmol/L ammonium acetate and 5 μmol/L methanephosphonic acid , with a flow rate maintained at 0. 35 ml/min . The mass spectrometry detection system utilized an electrospray ionization technique with negative ion mode for multiple reaction monitoring. Results The correlation coefficients of the standard curves for the six tricarboxylic acid cycle organic acid metabolites were all above 0. 996 within the quantitative range . The method ’s accuracy ranged from 97. 14% to 108. 26% , with inter-day and intra-day precision relative standard deviation be- tween 1 . 35% and 6. 73% . The matrix effect was between 93 . 29% and 107. 47% , and the extraction recovery rate ranged from 94. 82% to 112. 57% . Analysis of six tricarboxylic acid cycle organic acids in the liver , serum , and placenta of DEHP-exposed mice during pregnancy showed significant reductions in fumaric acid , malic acid , α-ke- toglutaric acid , cis-aconitic acid , and citric acid compared to the control group ( P < 0. 05) . Conclusion The HPLC-MS/MS method established in this study for detecting six tricarboxylic acid cycle organic acids in the serum , liver , and placenta of DEHP-exposed pregnant mice is stable , highly sensitive and selective . Prenatal DEHP expo- sure induced alterations in the levels of tricarboxylic acid (TCA) cycle organic acid metabolites in the liver , ser- um , and placenta of mice , suggesting that DEHP exposure during pregnancy may interfere with mitochondrial TCA cycle processes . These findings indicate potential value in the diagnosis and treatment of diseases associated with prenatal DEHP exposure .

abstract:
Alternative splicing (AS) is one of the important ways of post-transcriptional regulation , which can gen- erate multiple mRNA isoforms from a single gene . In recent years , many studies have shown that AS can occur in almost all types of tumors , and the abnormal expression of splicing factors is related to the disease progression of tumor patients , which may become a potential marker for judging tumor progression . Therefore , this review summa- rizes the role , molecular mechanism , clinical relevance and treatment response of AS in tumors . It is found that AS can participate in the progression of malignant tumors by regulating tumor invasion , metastasis , apoptosis , cell me- tabolism , and promoting tumor immune escape and treatment resistance , which provides a theoretical basis for the clinical application of AS . The precise identification of tumor-specific AS and the development of small molecule in- hibitors targeting specific AS isoforms may provide a new direction for precise and personalized cancer treatment.

abstract:
Opioids are commonly used drugs in the clinic , mainly targeting different subtypes of opioid receptors distributed in central and peripheral neurons . They are the preferred drugs for treating acute pain or cancer pain . Delta opioid receptor (DOR) agonists are opiate drugs that target the delta receptor. In recent years , they have re- ceived widespread attention due to their ability to improve ischemia/reperfusion injury in multiple organs such as the heart , brain , and liver , as well as their potential for treating mental and neurological disorders . This review summarizes the mechanism research of DOR agonists in treating a variety of diseases , prospects their clinical appli- cation prospects , and finds that DOR agonists may exert anti-ischemia/reperfusion injury effects on organs by re- ducing apoptosis and mitochondrial damage , and activating various signaling pathways; they also exhibit anti-in- flammatory , antidepressant , and therapeutic effects on neurodegenerative diseases , providing a new perspective for the application of DOR agonists in treating related diseases .

abstract:
Breast cancer is the most common malignancy among women worldwide , and its recurrence , metastasis , and drug resistance remain significant challenges in systemic treatment. Metabolic alterations are a hallmark of cancer , with breast cancer cells reprogramming glucose metabolism , lipid metabolism , and amino acid metabolism to adapt their nutrient acquisition pathways . This metabolic reprogramming enables cancer cells to meet the energy demands for rapid proliferation and adaption to the microenvironment of metastatic sites , which is closely associated with resistance to chemotherapy , targeted therapies , and immunotherapies . This review summarizes the features of metabolic remodeling in breast cancer , the mechanisms by which metabolic adaptation contributes to treatment re- sistance , and emerging individualized strategies targeting metabolic vulnerabilities . Particular focus is placed on en- ergy metabolism , cholesterol homeostasis , and glutamine utilization in relation to tumor invasion , metastasis , and drug resistance . A better understanding of metabolic adaptation and accurate monitoring of metabolic dynamics may offer new insights into metabolic therapies and improve clinical outcomes .