Objective To establish and clinically evaluate a capillary electrophoresis-based approach for expanded carrier screening. Methods We used capillary electrophoresis to detect 448 disease-causing variants among 24 genes associated with 20 diseases in 1 099 individuals. The detected variants were confirmed by alternative methods. Results The capillary electrophoresis results were totally consistent with those of alternative methods. Of the 1 099 individuals, 190 (17. 3 % , 109 / 1 099) were identified as carriers for at least one condition, and the most common disease carried by individuals was GJB2-related non-syndromic hearing loss. Of the 1 075 females, 8 (7. 4 ‰, 8 / 1 075) were identified as carriers for X-linked disorders. Conclusion We successfully established a capillary electrophoresis-based method for expanded carrier screening.

Objective To investigate the effect of long non-coding RNA ( lncRNA) homologous anomalous box gene A11 antisense RNA (HOXA11-AS) on oxidative low density lipoprotein (ox-LDL) induced vascular endothelial cells injury by targeting micrornA-766-3p (miR-766-3p). Methods Human umbilical vein endothelial cells (HUVEC) were treated with 100 μg / mL ox-LDL for 24 hours to establish a cell injury model. HUVEC were divided into the control ( con) group, ox-LDL group, ox-LDL + si-NC group, ox-LDL + si-HOXA11-AS group, ox-LDL + miR-NC group, ox-LDL + miR-766-3p group, ox-LDL + si-HOXA11-AS + anti-miR-NC group, and the ox-LDL + si-HOXA11-AS + anti-miR-766-3p group. Expression of HOXA11-AS and miR-7666-3p was assessed by RT-qPCR. Cell apoptosis was detected using flow cytometry. The kits detected LDH release and intracellular SOD activity. ELISA method was used to calculate the levels of TNF-α and IL-1β in the culture medium. Dual luciferase reporter assay was used to confirm the targeting relationship between HOXA11-AS and miR-766-3p. Results Compared with the control group, the HUVEC cell apoptosis rate, LDH release, HOXA11-AS expression, and the levels of TNF-α and IL-1β in the culture medium of the ox-LDL group were significantly increased (P< 0. 05), the SOD activity and the miR-766-3p expression level were significantly reduced (P< 0. 05). Compared with the ox-LDL + si-NC group, the HUVEC cell apoptosis rate, LDH release, and the levels of TNF-α and IL-1β in the culture medium of the ox-LDL + si-HOXA11-AS group were significantly reduced (P< 0. 05), the SOD activity was significantly increased ( P < 0. 05). Compared with the ox-LDL + miR-NC group, the HUVEC cell apoptosis rate, LDH release, and the levels of TNF-α and IL-1β in the culture medium of the ox-LDL + miR-766-3p group were significantly reduced ( P < 0. 05), the SOD activity was significantly increased ( P < 0. 05 ). miR-766-3p is the a target of HOXA11- AS. Compared with the ox-LDL + si-HOXA11-AS + anti-miR-NC group, the HUVEC cell apoptosis rate, LDH release, and levels of TNF-α and IL-1β in the culture medium of the ox-LDL + si-HOXA11-AS + anti-miR-766-3p group were significantly increased (P< 0. 05), and the SOD activity was significantly decreased (P< 0. 05). Conclusion Silencing HOXA11-AS inhibits ox-LDL-induced vascular endothelial cell apoptosis, oxidative stress and inflammation by up-regulating the expression ofmiR-766-3p. 

Objective To explore the expression of miR-144 in the peripheral blood serum and monocytes from patients with chronic obstructive pulmonary disease (COPD) and its clinical significance. Methods Fifty-eight COPD patients at the stable period admitted to our hospital from March 2020 to June 2021 were selected as the observation group, and 45 healthy patients during the same period were selected as the control group. The general clinical data of all patients were collected. Real-time fluorescent quantitative PCR ( qRT-PCR) was used to detect the expression level of miR-144 in the peripheral blood serum and monocytes of all patients. The relationship between the expression level of miR-144 in the peripheral blood serum and monocytes and the lung function in patients with COPD was analyzed. Pearson correlation was used to analyze the correlation between the expression level of miR-144 in the peripheral blood serum and monocytes of COPD patients and the forced expiratory volume in first second (FEV1). ROC curve was used to analyze the diagnostic value of miR-144 expression levels in the peripheral blood serum and monocyte for COPD. Results The expression level of miR-144 in the peripheral blood serum and monocytes of COPD patients was significantly higher compared with the control group, (P < 0. 05). The expression level of miR-144 in the peripheral blood serum and monocytes of COPD patients increased with the aggravation of lung function, and the difference was statistically significant (P< 0. 05). The results of Pearson correlation analysis showed that the expression level of miR-144 in the peripheral blood serum and monocytes of COPD patients was negatively correlated with FEV1 ( r = - 0. 517, r = - 0. 463, P< 0. 05). ROC curve analysis results showed that the areas under the ROC curve (AUCs) of miR-144 levels in the peripheral blood serum and monocyte for the diagnosis of COPD were 0. 857 and 0. 849, with the cut-off values to be 1. 371 and 1. 890, the sensitivities to be 84. 50 % and 84. 50 % , and the specificities to be 80. 00 % and 88. 90 % , respectively. Conclusion The expression level of miR-144 in the peripheral blood serum and monocyte of patients with COPD is significantly up-regulated, they participate in the regulation of the development of the disease, and have a certain diagnostic value for the occurrence of COPD.

Objective To explore the effect of lncRNA CERS6-AS1 on the biological behavior of glioma cells and its possible mechanism. Methods qRT-PCR method was used to detect the expression levels of CERS6-AS1 and miR-138-2-3p in glioma tissues and adjacent tissues. Pearson method was used to analyze the correlation between CERS6-AS1 and miR-138-2-3p expression in glioma tissues. Human glioma cells T98G were cultured in vitro, and then were transfected with si-NC, si-CERS6-AS1, miR-NC, miR-138-2-3p mimics, si-CERS6-AS1 and anti-miR-NC, si-CERS6-AS1 and anti-miR-138-2-3p. The CCK-8 method, plate colony formation experiment, and transwell assay were used to detect the cell capabilities of proliferation, clone formation, migration and invasion. The dual luciferase reporter gene experiment was used to verify the targeting relationship between CERS6-AS1 and miR-138-2-3p. Results Compared with the adjacent tissues, the expression level ofCERS6-AS1 in glioma tissues was increased (P< 0. 05), while the expression level of miR-138-2-3p was decreased (P< 0. 05). CERS6-AS1 was negatively correlated with miR-138-2-3p (r = - 0. 8899, P< 0. 001). CERS6-AS1 could target the expression of miR-138-2-3p. The cell viability, the number of colony forming cells, the number of migration and invasion cells in the si-CERS6-AS1 group were lower than those in the si-NC group (P< 0. 05). The cell viability, the number of colony formed cells, the number of migrated and invasive cells in the miR-138-2-3p group were less than those in the miR-NC group (P< 0. 05). The cell viability, the number of clone formed cells, the number of migrated and invasive cells in the si-CERS6-AS1 + anti-miR-138-2-3p group were all higher than those in the si-CERS6-AS1 + anti-miR-NC group (P< 0. 05). Conclusion Interference of the expression of CERS6-AS1 could inhibit the proliferation, clone formation, migration and invasion of glioma cells by regulating miR-138-2-3p.

Objective To explore the molecular mechanism of lncRNA PSMA3-AS1 on regulating the proliferation, migration and invasion of cervical cancer cells. Methods Collect cancer tissue and paracancerous tissue specimens of 42 patients with cervical cancer admitted to the Second Affiliated Hospital of Xi’an Medical University from March 2020 to June 2021. The qRT-PCR method was used to detect the expression levels of lncRNA PSMA3-AS1, miR-3619-5p in cervical cancer tissues, adjacent tissues, human cervical epithelial immortalized cells H8, and human cervical cancer cell lines SiHa, HeLa, and Cashi. Taking SiHa cells as the research object, they were randomly grouped into 6 groups: si-NC group, si-lncRNA PSMA3-AS1 group, miR-NC group, miR-3619-5p group, anti-miR-NC + si-lncRNA PSMA3-AS1 group, anti-miR-3619-5p + si-lncRNA PSMA3-AS1 group. The proliferation, migration and invasion abilities of SiHa cells in each group were assayed by MTT assay and transwell assay respectively. The dual luciferase reporter assay was used to determine the effect of miR-3619-5p overexpression on the luciferase activity of wild-type vector lncRNA PSMA3-AS1-WT and mutant vector lncRNA PSMA3-AS1-MUT. Western blotting was used to detect the protein expression levels of MMP2 and MMP9. Results The expression level of lncRNA PSMA3-AS1 in cervical cancer tissues was higher than that in the adjacent tissues (P< 0. 01), and the expression level of miR-3619-5p was lower than that in the adjacent tissues (P< 0. 01). Compared with the H8 cells, the expression level of lncRNA PSMA3-AS1 in SiHa, HeLa, and Caski cells was increased (P < 0. 01), while the expression level of miR-3619-5p was decreased (P < 0. 01). Compared with the si-NC group, the cell viability, and the protein expression levels of MMP2, MMP9 in the si-lncRNA PSMA3-AS1 group were decreased (P< 0. 05), and the numbers of migrated and invaded cells were decreased (P < 0. 05). Compared with the miR-NC group, the cell viability and the protein expression levels of MMP2, MMP9 in the miR-3619-5p group were decreased (P< 0. 01), and the numbers of migrated and invaded cells were decreased (P < 0. 01). Overexpression of miR-3619-5p could inhibit the luciferase activity of lncRNA PSMA3-AS1-WT (P< 0. 01), but failed to affect the luciferase activity of lncRNA PSMA3-AS1-MUT. Compared with the anti-miR-NC + si-lncRNA PSMA3-AS1 group, the cell viability, and the protein expression levels of MMP2, MMP9 in the anti-miR-3619-5p + si-lncRNA PSMA3-AS1 group were increased (P< 0. 01), and the numbers of migrated and invaded cells were increased (P< 0. 01). Conclusion Interfering with the expression of lncRNA PSMA3-AS1 could reduce the proliferation, migration and invasion of cervical cancer cells by targeting miR-3619-5p.

Objective To analyze the immune related lncRNAs that affect the prognosis of colon cancer, and construct a related prediction model for the prediction of the prognosis of colon cancer patients. Methods Download the lncRNA expression profile of colon cancer in the TCGA database, and the data was standardized by TPM to analyze the differential expression of all lncRNAs. The KNN method was used to supplement the missing values. Immune-related lncRNAs were extracted and identified of by co-expression method, and then LASSO regression analysis was performed on the top 100 differentially expressed lncRNAs. Then the single-factor and multi-factor COX regression analysis were performed. Finally, based on the relationship between the lncRNA risk score and the gene expression, the risk factor association chart, the KM curve and the ROC curve for the evaluation of the value of the predicted model were constructed by using the ggplot2 package of R 4. 0. 2 statistical software. Results Through the analysis of differencial expression, a total of 2 258 lncRNAs were found to be differentially expressed in the cancer and paracancerous tissues, of which 1 648 were up-regulated and 610 were down-regulated. The top 100 differentially expressed immune related lncRNAs were selected for LASSO regression analysis, and a total of 12 lncRNAs were screened out. Univariate and multivariate COX regression analysis showed that AC092723. 1, AC007182. 1 and AC004947. 1 were significantly related to the prognosis. Using R 4. 0. 2 statistical software to construct a prognostic risk factor association chart. The ROC curve showed that the predictive values for the prediction of 1, 3 and 5 years were high, and the AUC were 0. 79 (95 % CI: 0. 67-0. 91), 0. 78 (95 % CI: 0. 66-0. 9), 0. 7 (95 % CI: 0. 51-0. 9), respectively. Conclusion This study uses the TCGA public database for the bioinformatics analysis and constructs a prognostic model that shows high predictive values. In addition to certain clinical significance, it also provides certain directions for the future researches of lncRNAs in colon cancer.

Objective To investigate the expression of IDO1 in different stages of lung squamous cell carcinoma and its relationship with prognosis. Methods Patients with lung squamous cell carcinoma who underwent radical surgery in the Department of thoracic surgery of Xingtai People’s Hospital Affiliated to Hebei Medical University from January 2017 to January 2019 were included. Intraoperative cancer tissue samples were taken. The expression level of IDO1 was detected by immunohistochemical staining, and the patients were followed up after operation. Results IDO1 protein was expressed in lung squamous cell carcinoma, and the number of positive cells in clinical stages Ⅱ ~ Ⅲ was significantly higher than that in stage I. By the end of the follow-up, 28 of the 30 patients had survived, 2 died, 22 had stable disease (73. 3 % ), and 8 had disease progression (26. 7 % ). The progression free survival (PFS) ofido1high expression group andido1low expression group were 26 months and 40 months respectively (P < 0. 05). The PFS of higher clinical stage and lower clinical stage were 33 months and 40 months respectively (P < 0. 05). Conclusion The expression level of IDO1 in lung squamous cell carcinoma was related to the clinical stage. The higher the clinical stage, the higher the expression level, and the worse the prognosis of patients. IDO1 expression level can be used to guide the clinical staging and prognosis in lung squamous cell carcinoma. 

Objective To explore the relationship between the expression of sex determining gene box 9 (SOX9), Beclin1 and the prognosis in patients with oral cancer. Methods A total of 38 oral cancer patients admitted to the Baoding First Central Hospital of Hebei Province from May 2017 to May 2018 were selected, and 38 oral cancer tissue specimens and 30 adjacent tissue specimens were selected to detect the expression levels of SOX9 and Beclin1 in the patients. The relationship between the expression of SOX9 and Beclin1 and the clinicopathological factors was analyzed, and the risk factors affecting the prognosis of oral cancer were identified by COX. The relationship between SOX 9 and Beclin1 and the prognosis of oral cancer was analyzed. Results The positive expression rates of SOX9 and Beclin1 in the oral cancer tissues were significantly higher than those in the adjacent normal tissues (P< 0. 05). The positive expressions of SOX9 and Beclin1 were not associated with age and gender (P > 0. 05), but were correlated with tumor stages, degree of differentiation and lymphatic metastasis (P< 0. 05). Stepwise Cox regression analysis was used to analyze the prognostic factors of oral cancer. The results showed that tumor stages, lymph node metastasis, SOX9, and Beclin1 expressions were independent risk factors affecting the prognosis of oral cancer patients (P< 0. 05). Patients with positive SOX9 expression had a progression-free survival of 10- 18 months, with an average of (12. 5 ± 2. 4) months, and patients with negative SOX9 expression had a progression-free survival of 14-23 months, with an average of (17. 5 ± 3. 1) months. Patients with positive expression of Beclin1 had a progression-free survival of 11-18 months, with an average of (13. 7 ± 2. 4) months, and patients with negative Beclin1 expression had a progression-free survival of 13-22 months, with an average of (15. 7 ± 3. 1) months. Patients with positive expression of SOX9 and Beclin1 had a significantly lower progression-free survival than those with negative expression (c²=7.969、3.315，P＜0.05）. Conclusion SOX9 and Beclin1 were positively expressed in patients with oral cancer, and their expression was closely related to the prognosis. The patients with higher expressions of SOX9 and Beclin1 in oral cancer had worse prognoses.

Objective To analyze the differentially expressed genes (DEGs) and miRNAs of cervical cancer based on bioinformatics, and further verify the differentially expressed genes and proteins, in order to find potential biomarkers and therapeutic targets. Methods Cervical cancer related data were obtained from TCGA database, and DEGs and differential miRNAs were screened by edgeR algorithm. The mRNA-miRNA co-expression network was constructed using Cytoscape3. 8. 2 software. GO enrichment analysis and KEGG enrichment analysis were performed for DEGs and target genes of differential miRNA predicted by miRWalk website using DAVID software. DEGs was further verified by qPCR and Western blotting. Results A total of 149 up-regulated and 171 down-regulated DEGs, 46 up-regulated differential miRNAs and 64 down-regulated differential miRNAs were screened out. The enrichments of DEGs and miRNA target genes were consistent in cell composition, and they were all enriched in cytoplasm, nucleus and cytoplasm. However, co-expression network found no significant regulatory relationship between DEGs and differential miRNAs. Therefore, we subsequently focused on the verification of DEGs, and verified TCEAL6, CLEC3B, LMOD1 and CNN1 with relatively significant differential expression. QPCR showed that the expression levels of CNN1 in cervical cancer were significantly reduced, which was in line with expectations. Western blotting analysis of CNN1 also showed that it was lowly expressed in cervical cancer. Conclusion In this study, DEGs and differentially expressed miRNAs of cervical cancer were screened and analyzed based on the TCGA database, and TCEAL6, CLEC3B, LMOD1 and CNN1were further verified. The results showed that they were significantly under-expressed in cervical cancer, which is expected to become biomarkers of cervical cancer

Objective To study the role and mechanism of Helicobacter pylori (HP) infection in regulating the esophageal cancer cell apoptosis through MDM2 / P53 pathway. Methods The resected esophageal cancer tissues were collected and the HP infection, the expression levels of MDM2 and P53 were detected. Esophageal cancer TE-1 cell line was cultured and intervened with HP infection, normal saline ( NS ) or ubiquitin proteasome inhibitor PS341, negative control (NC) siRNA or MDM2 siRNA were transfected into the cells. Cell viability, apoptosis rate, the expression level of MDM2 and P53 and the ubiquitination level of P53 were detected. Results The expression level of MDM2 in the HP positive esophageal cancer tissues was higher than that in the HP negative esophageal cancer tissues, and the expression level of P53 was lower than that in the HP negative esophageal cancer tissues (P < 0. 05). The cell viability, MDM2 expression level and the P53 ubiquitination level in TE-1 cells of the HP group were higher than those in the control group. The apoptosis rate and P53 expression level were lower than those in the control group. The expression level of P53 in TE-1 cells of the PS341 + Hp group was higher than that in the NS + Hp group (P< 0. 05). The cell viability, MDM2 expression level and P53 ubiquitinated level in TE-1 cells of the si-MDM2 + Hp group were lower than those in the si-NC + Hp group, and the apoptosis rate and P53 expression level were higher than those in the si-NC + Hp group (P< 0. 05). Conclusion The inhibitory effect of HP infection on apoptosis of esophageal cancer cells is related to the increasing of the expression level of MDM2 and promotion of the ubiquitination and degradation of P53. 

Objective To explore the effect of chordin-like1 (CHRDL1) on the progression of colorectal cancer ( CRC). Methods The relationship between CHRDL1 and CRC progression, prognosis was analyzed by the database. SW480 and HT29 cell lines with CHRDL1 overexpression were constructed. The cell proliferation, migration and invasion were assayed. The tumor growth was detected byin vitrotumor transplantation assay. The mechanism of CHRDL1 overexpression on migration and invasion of CRC cells was analyzed. Results The expression of CHRDL1 was up-regulated in CRC progression and could affect the prognosis. The overexpression of CHRDL1 could promote cell proliferation, migration, invasion and growth of the transplanted tumors. Its overexpression could also promote the enhanced expressions of Vimentin, N-Cadherin and snail, and inhibit the expression of E-Cadherin. There was an interaction between CHRDL1 and snail. Interfering with snail siRNA could reverse the trend of the above indexes. Conclusion CHRDL1 can promote the proliferation of CRC cells, it can also promote the epithelial mesenchymal transformation of CRC cells and the cell migration and invasion by Snail.

Objective To explore the potential mechanism of HBV HBx protein on liver cancer cell proliferation regulation. Methods After HBx overexpression, the proliferation of HEPG2 liver cancer cells was detected. HBx was immunomotized co-precipitated in the Flag-HBx overexpressed HEPG2 cells, and then protein mass spectrometry was performed, the SCORE threshold was set to be greater than 50, and the Coverage threshold was set to be greater than 10. In HBx overexpressed HEPG2 cells, siRNAs were used to knock down the corresponding genes identified by the mass spectrometry, and the proliferation of cells was detected. Results The proliferation level of HEPG2 cells was increased after the expression of Flag-HBX, ( P < 0. 05). Compared with the control group, the proliferation of HEPG2 cells was decreased after the knock-down of Mcur1 (P< 0. 05). Compared with the control group, the overexpression of Mcur1 and HBx at the same time can promote the proliferation of HEPG2 (P< 0. 05), and the overexpression of MCUR1 only has no effect on the proliferation of HEPG2. Compared with the HBx group, the proliferation of HEPG2 cells with overexpressed HBx and lowly expressed MCUR1 was decreases (P< 0. 05). Compared with the control group, the ROS level of HEPG2 after HBx overexpression was increased (P < 0. 05), and it had no significant change in HEPG2 with overexpression of HBx and knock-down of MCUR1 at the same time. Compared with the control group and the HBx group, the overexpression of MCUR1 and HBx at the same time can increase the ROS level of HEPG2 (P < 0. 05). Compared with the HBx group, the proliferation of HEPG2 was decresed when HBx was overexpressed and the MCUR1 was knock down (P< 0. 05), however, this decreasing could be restored by H2O2 treatment. Compared with the control group, the positioning of nuclear transcription factor Nrf2 was increases in the HBx overexpressed group (P< 0. 05), while it had no significant change in cells with HBx overexpression and MCUR1 knock-down at the same time. Compared with the control group, the activated form of NOCD1 in the nucleus was increased, and it had no significant change after using ML385 in HBx overexpressed cells. Compared with the HBx group, the proliferation level of HEPG2, HEPG2 after ML385 expressed HBX at the same time as HBX (P<0. 05). Compared with the HBX group, the proliferation of HEPG2 was decreased after treatment of NOTCH inhibitor IMR-1 or ML385. Conclusion HBx promotes HepG2 cell proliferation through the MCUR1/ ROS/ Nrf2/ Notch axis

Objective To investigate the effect of Phillyrin on lipid metabolism disorder and oxidative stress in obese rats. Methods The obese rats were fed with high-fat diet. The rats were randomly divided into 6 groups: the control group, the model group, the phillyrin 5 mg / kg group, the phillyrin 10 mg / kg group, the phillyrin 20 mg / kg group and the metformin group. The weight of rats was measured. A tape measure is used to measure the nose-to-anal length and the Lee’s index was calculated. The blood glucose level was determined by blood glucose meter. The lipid accumulation in rats was assayed by Oil red O staining. The levels of TC, TG, LDL-C and HDL-C were measured by automatic biochemical analyzer. The levels of ALT, AST, ALP, SOD, MDA and GSH-Px were detected by the kit. The protein expression levels of p-Nrf2, Nrf2 and HO-1 were detected by Western blotting. Results Compared with the control group, the Lee’s index, the weight gain and the blood glucose level in the model group were significantly increased, the levels of TC, TG, LDL-C, ALT, AST and ALP were significantly increased, the level of HDL-C was significantly decreased, the SOD and GSH-Px content were significantly decreased, the MDA content was significantly increased. The protein expression levels of p-Nrf2 / Nrf2 and HO-1 were significantly decreased (P< 0. 05). Compared with the model group, the Lee’s index and the blood glucose level in the phillyrin 10 and 20 mg / kg groups and the metformin group were decreased, the degree of weight gain and lipid accumulation were significantly improved in rats, the levels of TC, TG, LDL-C ALT, AST and ALP levels were decreased, the level of HDL-C was increased, the SOD and GSH-Px content were increased, the MDA content was decreased, and the expression levels of p-Nrf2 / Nrf2 and HO-1 were increased (P < 0. 05). Conclusion Phillyrin can relieve the oxidative stress and lipid metabolism disorder in obese rats, which possibly through the activation of Nrf2 / HO-1 pathway.

Transient channel receptor potential cation 6 (TRPC6) is one of the most essential proteins of the slit diaphragm (SD) on podocytes in the kidney. It interacts with the actin, surface proteins, and other SD components to keep the podocytes functioning normally. The over activation of TRPC6 causes intracellular calcium overload, which injures or destroys podocytes. It is now known that a pathogenic mutation in the TRPC6 gene might cause focal segmental glomerulosclerosis. TRPC6 has been linked to the onset and progression of proteinuric kidney disease in recent researches. However, there is no particular targeted medication for the clinical therapeutics. The molecular mechanisms of podocytes injuries due to TRPC6, as well as the relationship between genetic mutations in TRPC6 and clinical phenotypes are discussed in this review

Lung cancer remains the leading cause of cancer-related death world-wide, with a 5-year survival rate of only about 16 % . As the main host cells in the tumor microenvironment (TME), cancer-associated fibroblasts (CAFs) play an important regulatory role in the occurrence and development of lung cancer. Chemokines derived from CAFs can activate downstream signal pathways by binding with receptors, and play roles in tumor proliferation, invasion, metastasis, angiogenesis, drug resistance, radiation resistance, immune evasion and so on. This review summarizes the latest research progress of chemokines derived from CAFs in lung cancer, discusses the potential of using them in the diagnosis and treatment of lung cancer, and provides new targets and directions for the future diagnosis and treatment of lung cancer.