abstract:
Objective To investigate the effects of adjuvant arthritis(AA) on renal function and the expression of organic anion transporter 3(OAT3) in rats. Methods The AA model was establishedviaintradermal injection of Freund′s complete adjuvant into the right hind toe of the rats. Blood and urine were collected at fixed time points to observe the dynamic changes of renal injury indicators. The kidney injury was observed by HE staining. Immunohistochemistry and Western blot were used to detect the expression levels of OAT3 protein and inflammatory factors interleukin-6(IL-6), interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) in renal cortex. Results Compared with the normal group, the arthritis index score, the secondary paw swelling, the number of joint swelling, and the whole body score of AA rats significantly increased(P<0.05). The X-ray results showed that the AA model rats had soft tissue edema and bone deformity. Biochemical indicators showed that the renal injury indicators of AA rats were significantly abnormal. The results of immunohistochemistry and Western blot showed that the expression level of OAT3 in renal cortex of AA rats significantly decreased, while the expression levels of inflammatory factors IL-6, TNF-α and IL-1β significantly increased(P<0.05). Conclusion The renal injury of AA rats may be related to the decreased expression level of OAT3 and the increased expression level of inflammatory factors in kidney.

abstract:
Objective To develop liver cancer-targeted nanoparticles(LA-CMGL) co-loaded with miR-145/camptothecin(CPT) and assess their targeting specificity, combined anti-tumor effects, and magnetic resonance imaging efficacy. Methods Laser scanning confocal microscopy and flow cytometry were utilized to evaluate the targeted uptake of lactobionic acid-modified and unmodified nanoparticles by HepG2 cells and HepaRG cells; quantitative polymerase chain reaction(qPCR) was employed to assess miR-145 content in tumor cells and tissues. The cytotoxicity of CPT, LA-CPT-LNPs and LA-CMGL on HepG2 cells were assessed using the CCK-8 assay. qPCR was also used to evaluate the effect of CPT, LA-CPT-LNPs and LA-CMGL on apoptosis of HepG2 cells. MRI was performed to measure the relaxation rate of LA-CMGL and evaluate its targeting imaging effect on liver cancer cells. Results The uptake rate of LA-CMGL by HepG2 cells surpassed that of CMGL significantly. The relative miR-145 content in liver cancer cells and mouse liver cancer tissues in the LA-CMGL group was markedly higher compared to free miR-145 and CMGL groups(P<0.001). The apoptosis rate of HepG2 cells in the LA-CMGL group exceeded that in the CMGL group and CPT group(P<0.01). At the same Gd3+concentration, the relaxation rate of LA-CMGL significantly surpassed that of Gd-DOTA, and the MRI signal of LA-CMGL in HepG2 cells markedly increased compared to CMGL and Gd-DOTA. Conclusion LA-CMGL exhibits promising liver cancer-targeted delivery, combined anti-tumor effects, and MRI liver cancer cell-targeted imaging, offering a novel avenue for combined drug/gene therapy for liver cancer.

abstract:
Objective To investigate the expression of long non-coding RNA(LncRNA) PICSAR in ovarian cancer, and explore the effects of LncRNA PICSAR on the proliferation, migration, invasion and apoptosis of ovarian cancer cells as well as its possible mechanism of action. Methods The expression levels of LncRNA PICSAR in ovarian cancer tissue and cell line A2780, OVCAR-3, HO-8910 and normal ovarian tissue and cell line IOSE386 were detected by fluorescence quantitative PCR. Ovarian cancer cell lines with the highest expression of LncRNA PICSAR were divided into control group and experimental group, and transfected with negative control small interfering RNA(siRNA-NC) or PICSAR knockout small interfering RNA(siRNA-PICSAR) by liposome transfection technique, respectively. The effects of LncRNA PICSAR knockdown on the invasion, migration, proliferation and apoptosis of ovarian cancer cells were analyzed by cell counting assay(CCK-8), clonogenic assay, scratch assay, transwell assay and flow cytometry and so on. The expression levels of autophagy related proteins and apoptosis-related proteins in each group were determined by Western blot. Ovarian cancer cells were treated with rapamycin and hydroxychloroquine as autophagy activator and inhibitor, and Western blot assay was used to detect apoptosis. Results The expression level of LncRNA PICSAR in ovarian cancer tissues was higher than that in normal ovarian tissues. Compared with IOSE386 cell line, LncRNA PICSAR expression levels in ovarian cancer cell lines HO-8910, OVCAR-3 and A2780 increased. Compared with the si-NC group, the proliferation, invasion and migration ability of ovarian cancer cells in si-PICSAR group decreased, and the apoptosis rate increased. The autophagy level of ovarian cancer cells in si-PICSAR group was lower than that in si-NC group. After transfection with siRNA-PICSAR, rapamycin activated autophagy to reduce apoptosis, while hydroxychloroquine inhibited autophagy to promote apoptosis. Conclusion LncRNA PICSAR is highly expressed in ovarian cancer tissues and cell lines, and the malignant biological behavior of ovarian cancer cells can be inhibited by knockout of LncRNA PICSAR. The knockdown of LncRNA PICSAR may promote the apoptosis of ovarian cancer cells by regulating autophagy.

abstract:
Objective To explore whether the locus coeruleus(LC) mediates the pain in mice with depressive-like behaviors and underlying mechanisms.Methods Male C57BL/6J mice were randomly divided into two groups,including control and chronic restraint stress(CRS) group.Novelty-suppressed feeding test,tail suspension test and forced swimming test were used to assess the formation of depressive-like behaviors and the establishment of CRS model.The mechanical pain threshold of mice was detected by Von Frey brush test.LC neuronal activity was detected by c-Fos immunofluorescence staining.Neuronal excitability in LC was detected using electrophysiology of isolated brain slices.The influence on pain and depressive-like behaviors was identified by chemogenetics in LC.Results Compared with the Control group,CRS mice displayed significant depressive-like behavior after 3 weeks of CRS(CRS 3W),and the model was successful;CRS 3W mice showed a significant decrease in mechanical pain threshold and developed mechanical pain sensitivity.A lot of c-Fos+cells of LC was observed in CRS 3W mice,90% of them were noradrenergic neurons.Compared with the Control group,the neuronal excitability of noradrenergic neurons decreased in LC of CRS 3W mice.Compared with the Control group,chemogenetic activation of the LC noradrenergic neurons significantly reduced mechanical pain sensitization in CRS 3W mice.Conclusion The excitability of noradrenergic neurons in LC of CRS 3W mice decreases,and chemogenetic activation of noradrenergic neurons in LC can relieve pain sensitization in CRS 3W mice.

abstract:
Objective To investigate the difference between imaging flow cytometry and immunofluorescence techniques for detecting Ahr entry in fibroblast-like synoviocytes. Methods Human-derived fibroblast-like synoviocytes line MH7A was cultured with 15% DMEM. The nucleation levels of Ahr in MH7A were detected by laser scanning confocal microscopy and quantitative imaging flow cytometry(ImageStreamX Mark II) in the blank control group, the Kyn group, and the Kyn+CH223191 group, respectively. The results of the two assay techniques were compared using nonparametric tests-correlation of the results. Results Compared with the blank control group, the Kyn and Kyn+CH223191 groups of MH7A cells increased Ahr nucleation ability as measured by immunofluorescence technique and imaging flow cytometry(P<0.05), and the three groups of results calculated by the two experimental methods were consistent, with the R2 values of 0.863 8, 0.928 7, and 0.901 8, respectively, which were statistically significant differences(P<0.05). Conclusion Compared with the operation and results of immunofluorescence experiments, imaging flow cytometry data processing is more complicated. However, the results are notably precise, avoiding the subjectivity of the experimenter and minimizing experimental inaccuracies.

abstract:
Objective To investigate the toxicity and mechanism of a novel Staphylococcus aureus with incomplete hemolytic phenotype(SIHP) filtrate on non-small cell lung cancer(NSCLC) HCC827 cell line.Methods The filtrates and diluents of novel SHHP and Staphylococcus aureus with complete hemolytic phenotype(SCHP) were coincubated with 1% human red blood cells and HCC827 cells.Hemoglobin release method was used to detect red blood cell toxicity,CCK-8 method was used to detect HCC827 cell activity.Ion selective electrode method,colorimetric method and immunofluorescence method were used to detect potassium(K+),lactate dehydrogenase(LDH) and interleukin-6(IL-6) concentrations in the incubation supernatants.Results The filtrate and diluent of novel SIHP could significantly damage the activity of HCC827 cells,causing K+and LDH leakage.The filtrate and 1:3 diluent of novel SIHP resulted in a decrease and an increase in IL-6 secretion in HCC827 cells,respectively.Compared with SCHP,the filtrate and diluent of novel SIHP had stronger red blood cell toxicity and more severe K+leakage in HCC827 cells.Conclusion The filtrate of novel SIHP can damage the cell membrane,lead to content release and kill NSCLC cells,with a stronger toxicity than SCHP.

abstract:
Objective To explore the microbiological characteristics ofStaphylococcus aureus(S.aureus) with novel incomplete hemolytic phenotype(SIHP). Methods Hemolytic phenotypes were detected and categorized by using the three-point inoculation method. A total of 11 novel SIHP and 33 randomly matchedS.aureuswith complete hemolytic phenotype(SCHP) were included. Antibiotic susceptibility test was performed using broth microdilution method. Coagulase test was performed with freeze-dried rabbit plasma. Catalase activity was detected by slide catalase test. Expression of hemolysin genes was detected by qRT-PCR. Toxicity to human red blood cells was assessed by microplate method. Microplate biofilm formation was measured using crystal violet staining method. Growth kinetic determination was performed through microcultivation assay. Results Compared with SCHP, the expression profiles of the four hemolysin genes(hla,hlb,hlc, andhld) in the new SIHP were different. The new SIHP had higher resistance rates to penicillin, oxacillin, gentamicin, quinolones, clindamycin, and trimethoprim-sulfamethoxazole. Furthermore, the new SIHP had stronger hemolytic toxicity, plasma coagulase activity, and biofilm formation ability. Additionally, the new SIHP grown faster in the logarithmic phase. Conclusion Taken together, the microbiological characteristics of the new SIHP are different from those of SCHP, including stronger antibiotic resistance and pathogenicity, which should be paid more attention by clinicians.

abstract:
Objective To explore the effects and action mechanism of miR-181c-5p on biological behaviors of prostate cancer cells. Methods The pathological relationship between BIRC5, miR-181c-5p and prostate cancer was analyzed based on prostate cancer data in TCGA database. The target binding site of miR-181c-5p and BIRC5 was analyzed by miRNA target gene prediction database, and was verified by double luciferase activity assay. The expression of BIRC5 protein in miR-181c-5p overexpression cells was detected by Western blot. The prostate cancer cells PC3 and DU145 were selected to construct cell line with miR-181c-5p overexpression(miR-181c-5p group) and its negative control(miR-NC group), and qRT-PCR verification was conducted. The cells proliferation [optical density at 450 nm site(OD450 nm)] was detected by CCK-8. Distribution of cell cycles and apoptosis rate were detected by flow cytometry. Expressions of proliferation and apoptosis related proteins were detected by Western blot. The cell line with miR-181c-5p/BIRC5 overexpression was constructed(miR-181c-5p+BIRC5 group). Cells growth, distribution of cell cycles, apoptosis rate and expressions of related proteins were detected by the above methods. Results The expression of BIRC5 was up-regulated in prostate cancer tissues, and it was higher in patients with high tumor invasion, lymph node metastasis and recurrence. Patients exhibiting high expression of BIRC5 demonstrated poor survival rates. The expression of miR-181c-5p was down-regulated in prostate cancer tissues. The level of miR-181c-5p was negatively correlated with BIRC5 level, and miR-181c-5p could inhibit BIRC5 expression. In PC3 and DU145, miR-181c-5p level in miR-181c-5p group was higher than that in miR-NC group(P<0.05); OD450 nmand percentage of S-phase cells were lower than those in miR-NC group(P<0.05), percentage of cells in G0/G1phase; apoptosis rate and expressions of BAX, caspase-3 and PARP proteins were higher than those in miR-NC group(P<0.05); expressions of CDK2, CCNB1 and BCL-2 proteins were lower than those in miR-NC group(P<0.05). The expression of BIRC5 protein and OD450 nmin miR-181c-5p+BIRC5 group were higher than those in miR-181c-5p group(P<0.05), percentage of cells in G0/G1phase was lower than that in miR-181c-5p group(P<0.05); percentage of S-phase cells was higher than that in miR-181c-5p group(P<0.05); apoptosis rate was lower than that in miR-181c-5p group(P<0.05); expressions of CDK2, CCNB1 and BCL-2 proteins were higher than those in miR-181c-5p group; expressions of BAX, caspase-3 and PARP proteins were lower than those in miR-181c-5p group. Conclusion miR-181-5p can inhibit the proliferation of human prostate cancer cells by targeting BIRC5, block cells in G0/G1phase and promote cells apoptosis.

abstract:
Objective To analyze the clinical significance of elevated CHMP2B expression in pancreatic cancer, and to explore the oncogenic mechanism of CHMP2B in pancreatic cancer. Methods A transcriptomic analysis conducted on the GEPIA website revealed CHMP2B expression in over 20 types of malignant tumors. The relationship between CHMP2B expression and disease-free survival and overall survival of pancreatic cancer was analyzed. The correlation between CHMP2B, MAP1LC3B and P62 expression was analyzed by TCGA database. The overexpression plasmid of CHMP2B factor was transfected with Lipofectamine 2000 in MIA PaCa-2 cells, and the overexpression of CHMP2B factor in MIA PaCa-2 was verified by Western blot. The effect of overexpression of CHMP2B on tumorigenic ability of MIA PaCa-2 cells was verified by clonal formation experiment. The effect of overexpression of CHMP2B on proliferation, migration and invasion of MIA PaCa-2 cells was verified by CCK-8 and Transwell experiments. The localization of CHMP2B factor was detected by immunofluorescence staining. The correlation between CHMP2B, MAP1LC3B and P62 expression was analyzed by TCGA database. The expression of MAP1LC3B and P62 in CHMP2B overexpression was verified by qPCR. Results The expression of CHMP2B factor increased in pancreatic cancer(P<0.01). Overexpression of CHMP2B factor was associated with overall survival and disease-free survival(P<0.01). The high expression of CHMP2B promoted the tumorigenesis of MIA PaCa-2 cells in pancreatic cancer, and also promoted the proliferation, migration and invasion of pancreatic cancer. Immunofluorescence staining showed that CHMP2B was distributed in both nucleus and cytoplasm and correlated with vesicle secretion. Moreover, CHMP2B was positively correlated with the expression of autophagy marker MAP1LC3B while inhibiting the expression of P62. Conclusion The high expression of CHMP2B in clinical pancreatic cancer tissue samples is consistent with the high expressionin vitropancreatic cancer cell lines. CHMP2B is localized in both nucleus and cytoplasm, involving in vesicle transport and lysosomal autophagy. In the pancreatic cancer cell line MIA PaCa-2, CHMP2B is positively correlated with the expression of autophagy marker MAP1LC3B and inhibits the expression of P62. It is concluded that CHMP2B is related to autophagy in promoting the malignant degree of pancreatic cancer. CHMP2B meets the high nutritional metabolic requirements of pancreatic cancer cells by regulating vesicular transport and autophagy, thus promoting the malignant biological behavior of pancreatic cancer.

abstract:
Objective To investigate the effect of DEAD-box RNA helicases(DDX5 helicase) on the proliferation, apoptosis and differentiation of leukemia cell line K562 cells. Methods Data from the Gene Expression Profiling Interactive Analysis(GEPIA) cancer gene database was utilized to analyze the expression of DDX5 mRNA in tissues of leukemia patients. Survival curve analysis was conducted to assess the relationship between DDX5 mRNA expression levels and the prognosis of leukemia patients. Small interfering RNA(siRNA) was used for transient transfection of K562 cells to knock down DDX5. Real-time quantitative PCR(RT-PCR) was employed to verify the silencing effect, and Western blot was used to detect protein expression levels. CCK-8 assay was conducted to examine cell proliferation capability, and Western blot and immunofluorescence were utilized to detect the expression levels of cell proliferation-related proteins. Flow cytometry was used to detect cell apoptosis, and the expression levels of cell differentiation-related proteins were assessed by flow cytometry and Western blot. Lastly, the effects of DDX5 silencing on the expression levels of P21 and P53 proteins were examined by RT-PCR and Western blot. Results GEPIA database analysis revealed that the expression level of DDX5 in human leukemia bone marrow tissues was significantly higher than that in healthy individuals, and patients with low DDX5 expression had longer survival time.In vitroexperiments demonstrated that knocking down DDX5 significantly inhibited the proliferation of K562 cells. Flow cytometry results indicated that DDX5 silencing promoted apoptosis and induced cell differentiation by enhancing the expression of CD11b and CD14 proteins. Furthermore, silencing DDX5 up-regulated the expression levels of P21 and P53 proteins. Conclusion DDX5 may potentially inhibit the proliferation of leukemia cell line K562, promote apoptosis, and induce differentiation by targeting P53.

abstract:
Objective This study examined the influence of c-Myc regulated long noncoding RNA TBL1XR1-5(lnc-TBL1XR1-5) on the progression of oral squamous cell carcinoma(OSCC). Methods Based on the expression of c-Myc, bioinformatics analysis was performed on head and neck squamous cell carcinoma samples in theTCGA database, and lnc-TBL1XR1-5 with closely related c-Myc expression was screened.The effects of c-Myc overexpression and knockdown on lnc-TBL1XR1-5 were detected. Expression relationship of c-Myc and lnc-TBL1XR1-5 in OSCC and para-cancer tissues. Dual-luciferase reporter assays were used to verify the binding of c-Myc to the lnc-TBL1XR1-5 promoter region. RNA FISH were used to determine the localization of lnc-TBL1XR1-5 in OSCC cell lines. The effects of overexpression, knockdown of lnc-TBL1XR1-5 on migration, metabolism, and proliferation of OSCC cells were observed by quantitative real-time polymerase chain reaction(qRT-PCR), scratch tests, transwell assays, medium color and pH changes, cell counts, CCK-8 assay, and colony formation assays. Results It was found that c-Myc positively regulated the expression of lnc-TBL1XR1-5 in OSCC cell lines and tissues. The binding of c-Myc to the lnc-TBL1XR1-5 promoter region was verified by dual-luciferase reporter assays. RNA FISH showed that lnc-TBL1XR1-5 was localized in the nuclei in OSCC cells. Overexpression of lnc-TBL1XR1-5 promoted migration, metabolism and proliferation in OSCC cell lines, while knockdown of it had the opposite effect. Conclusion c-Myc positively regulates lnc-TBL1XR1-5 in OSCC. Lnc-TBL1XR1-5 affect the migration, proliferation, and metabolism of OSCC by influencing the tumor microenvironment.

abstract:
Objective To explore the role of mitochondrial autophagy in ovarian inflammation associated with polycystic ovary syndrome(PCOS) based on the NOD-like receptor thermoprotein domain-related protein 3(NLRP3) pathway. Methods Human ovarian granulosa cell line SVOG was treated with 25 nmol/L dihydrotestosterone(DHT) for 24 h to establish PCOS cell model. SVOG cells were transfected with adenovirus carrying NLRP3(Ad-NLRP3) and negative vector(Ad-EV) or NLRP3 shRNA(sh-NLRP3) and negative control(sh-NC) to overexpress or knockdown NLRP3. Mito-Tracker staining and GFP-LC3 staining were used to evaluate mitochondrial autophagy in cells. TUNEL staining, JC-1 staining and Mito-SOX staining were used to analyze the apoptosis, mitochondrial membrane potential and mitochondrial-derived superoxide production. 32 female BALB/c mice were randomly divided into three groups: control(Con) group, DHEA group, DHEA+sh-NC group and DHEA+sh-NLRP3 group, with 8 mice in each group. Except the control group, all other groups treated mice with dehydroepiandrosterone(DHEA) to establish PCOS mouse model. DHEA+sh-NLRP3 group and DHEA+sh-NC group were administrated with sh-NLRP3 or sh-NC encapsulated in lentivirus at a concentration of 1×109TU/mlviatail vein injection. The ultrastructure of mitochondria in ovarian tissue of mice in each group was observed by transmission electron microscope. Results Compared with DHT+sh-NC group, the level of NLRP3 of SVOG cells in DHT+sh-NLRP3 group decreased(P<0.05). The co-location of GFP-LC3 and mitochondria in SVOG cells in DHT+sh-NLRP3 group was higher than that in DHT+sh-NC group(P<0.05). Compared with DHT+sh-NC group, the number of TUNEL positive cells and Mito-SOX fluorescence density of SVOG cells in DHT+sh-NLRP3 group decreased, and the ratio of polymer JC-1 to monomer JC-1 increased(P<0.05). Compared with Con+Ad-EV group, the level of NLRP3, the number of TUNEL-positive cells and the fluorescence density of mito-SVOG in Con+Ad-NLRP3 group increased(P<0.05), and the co-location level of GFP-LC3 and mitochondria decreased; the ratio of polymer JC-1 to monomer JC-1 decreased(P<0.05). Compared with the control group, TUNEL positive cells, relative ROS intensity and percentage of damaged mitochondria in the ovarian tissue of mice in DHEA group increased(P<0.05). Compared with DHEA+sh-NC group, TUNEL positive cells, relative ROS intensity and percentage of damaged mitochondria in DHEA+SH-NLRP group decreased(P<0.05). Conclusion Inhibition of mitochondrial autophagy induced by activation of NLRP3 leads to mitochondrial dysfunction and promotes mitochondrial-related apoptosis in GCs. Knockdown of NLRP3 is beneficial to mitochondrial homeostasis and improves the resistance of GCs to oxidative stress injury, thus promoting the recovery of PCOS.

abstract:
Objective To explore the effect of miR-296-3p on hepatic fibrosis induced by bile duct ligation(BDL) in rats. Methods 25 SD rats were randomly divided into sham group, model(BDL) group, NC adv group, miR-296-3p adv group and miR-296-3p sponge adv group, with 5 rats in each group. The pathological changes were observed in rat liver tissueviaHE, Masson and Sirius Red staining; the levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST) and total bilirubin(TBIL) in the serum of rat in each group were detected; the expression levels of miR-296-3p, interleukin(IL)-6, IL-1β, tumor necrosis factor(TNF)-α and smooth muscle actin(α-SMA), type I collagen(Col1A1), and connective tissue growth factor(CTGF) mRNA in rat liver tissue were detected by qRT-PCR; the expression levels of α-SMA, Col1A1 and CTGF proteins were detected by Western blot. Immunohistochemical staining(IHC) was performed to detect the expression of α-SMA in liver tissue. Target genes of miR-296-3p was predicted by bioinformatic analysis using the online database. Zinc finger and BTB domain-containing protein20(ZBTB20) mRNA and protein expression levels were detected. Results The pathological staining results showed that compared with sham group, a large number of infiltrated inflammatory cells and collagen deposition were observed in the liver tissues of rats in the BDL group and NC adv group. Compared with NC adv group, the inflammatory cells and collagen deposition decreased in the liver tissues of miR-296-3p adv group. However, in miR-296-3p sponge adv group, collagen product and inflammatory reaction increased. Compared with sham group, the contents of ALT, AST and TBIL in serum of rats in BDL group and NC adv group increased, the expression level of miR-296-3p decreased, the mRNA expression levels of IL-6, IL-1β, TNF-α increased, and the mRNA and protein expression levels of α-SMA, Col1A1 and CTGF increased(allP<0.05). Compared with the NC adv group, the contents of ALT, AST and TBIL in serum of rats in miR-296-3p adv group decreased, the expression level of miR-296-3p increased, the mRNA expression levels of IL-6, IL-1β and TNF-α, and the mRNA and protein expression levels of α-SMA, Col1A1 and CTGF in liver tissues decreased(allP<0.05). The results of miR-296-3p sponge adv group were opposite to those of miR-296-3p adv group(allP<0.05). The bioinformatics website predicted that ZBTB20 might be a candidate target gene of miR-296-3p. Compared with sham group, the expression of ZBTB20 mRNA and protein in the liver tissues of BDL group and NC adv group increased(P<0.05), and the expression of ZBTB20 in the liver tissues of miR-296-3p adv group decreased compared with NC adv group(P<0.05). However, the expression of ZBTB20 in liver tissues of miR-296-3p sponge adv group increased(P<0.05). Conclusion miR-296-3p expression decreases in BDL-induced hepatic fibrosis in rats, and miR-296-3p may inhibit hepatic fibrosis in BDL rats by targeting ZBTB20.

abstract:
Objective To investigate the effects of β-caryophyllene(BCP) on the browning of white adipose tissue in obese mice and the related mechanisms. Methods An obese mouse model was establishedviaintraperitoneal injection of a high-fat diet supplemented with propylthiouracil saline solution [14.4 mg/(kg·d)] in male Kunming mice. Obesity model mice were randomly divided into a model group(Model group) and a BCP administration group(BCP-50 group); normal diet mice were set up as a control group(Control group), with 8 mice in each group. BCP administration was given by gavage at a dose of 50 mg/kg once in the morning and once in the evening in the BCP-administered group, while the rest of the group was administered by gavage with aqueous solution of Tween 80 for 4 weeks. The oral glucose tolerance test was performed at the end of 4-week administration, and mice were executed after overnight fasting at the end of the experiment, and blood samples and adipose tissues were rapidly collected for subsequent experimental tests. The kit was used to detect serological-related indexes; hematoxylin-eosin staining was conducted to observe the morphology of adipose tissue; immunohistochemical staining was carried out to observe the expression of uncoupling protein 1(UCP1) in adipose tissue; Western blot was employed to detect expression of peroxisome proliferator-activated receptor γ coactivator1-α(PGC1α), peroxisome proliferator-activated receptor γ(PPARγ), UCP1 and cannabinoid receptor 2(CNR2) proteins in epididymal white adipose(eWAT). Results Compared with the model group, the body mass of obese mice in the BCP-50 group was significantly reduced(P<0.05), food intake was decreased(P<0.01), insulin resistance was improved(P<0.000 1), and the serum content of low-density lipoprotein cholesterol(LDL-C) and nonesterified fatty acid(NEFA) in the obese mice was significantly reduced(P<0.000 1 andP<0.01). Total cholesterol(TC), triglyceride(TG), and high-density lipoprotein cholesterol(HDL-C) contents did not change significantly. In addition, the adiposity coefficient and eWAT specific gravity of obese mice in the BCP-50 group were significantly decreased(P<0.05); the adipocytes in eWAT and BAT were reduced; and the expression of the UCP1 protein was significantly elevated(P<0.01 andP<0.05). In addition to UCP1, the expression levels of PGC1α, PPARγ, and CNR2 proteins in the eWAT of obese mice in the BCP-50 group were also significantly elevated(P<0.01,P<0.05, andP<0.001). Conclusion β-caryophyllene promotes white adipose tissue browning through up-regulating PPARγ/PGC-1α/UCP1 pathway expression, thus improving obesity.

abstract:
Objective To explore the effect of astilbin(astilbin-CS-NPs)-loaded chitosan nanoparticles(CS-NPs) on high glucose(HG)-induced ferroptosis of renal tubular epithelial cellsviaregulating mitogen-activated protein kinase 14(MAPK14)/heat shock protein 27(HSP27). Methods HK-2 cells were divided into the following groups: normal glucose(NG) group, HG group, HG+astilbin-CS-NPs(0, 5, 10, 20 mg/L) group, and under HG condition, pcDNA3.1-MAPK14 group, pcDNA3.1-MAPK14+astilbin-CS-NP group, si-HSP27 group, pcDNA3.1-MAPK14+si-HSP27 group, and pcDNA3.1-MAPK14+si-HSP27+astilbin-CS-NPs group. Cell viability was detected using CCK-8 assay, cell apoptosis was detectedviaTunel assay. Meanwhile, iron ion levels, lactate dehydrogenase(LDH) activity, glutathione(GSH) levels, and reactive oxygen species(ROS) levels were measured using assay kits. Rat model of diabetic kidney disease(DKD) was constructed and intervened with astilbin-CS-NPs to explore the effects of astilbin-CS-NPs on DKDin vivo. Results Compared with the NG group, the HK-2 cell viability in the HG group was significantly reduced, apoptosis increased, iron ion levels, LDH activity, and ROS levels were significantly elevated, while GSH levels significantly decreased(allP<0.05). Treatment with astilbin-CS-NPs significantly reversed the effects of HG on the biological behavior of HK-2 cells and ferroptosis-related indicators. Additionally, compared to the pcDNA3.1 group, the pcDNA3.1-MAPK14 group showed increased ferroptosis, which was improved by knocking down HSP27 or co-intervention with astilbin-CS-NPs.In vivoexperimental results showed that astilbin-CS-NPs could improve DKD rat kidney injury, inhibit iron ion levels and the expression of MAPK14/HSP27. Conclusion Astilbin-CS-NPs may improve HG-induced ferroptosis of renal tubular epithelial cellsviainhibition of MAPK14 and HSP27 expression.

abstract:
Objective To investigate the molecular mechanism underlyingPorphyromonas gingivalis(Pg)-induced autophagy in esophageal squamous cell carcinoma(ESCC). Methods After Pg infected KYSE70 cells and KYSE140 cells pretreated with siAtg7 or Chloroquin(CQ), Western blot was used to measure protein levels of Atg7, LC3-Ⅱ/LC3-Ⅰ, and p62; Immunofluorescent confocal imaging analysis was used to detect autophagosome and autolysosome; CCK-8 assay was used to test cell viability; Transwell assay was used for ESCC cell migration and invasion potentials. Likewise, miR-21-5p inhibitor, RASA1 overexpression plasmid, or U0126 were used to block miR-21-5p/RASA1/ERK signaling pathway prior to Pg infection, followed by the aforementioned methods. In addition, immunohistochemistry was used to examine Pg abundance and LC3 protein levels, and RT-PCR was used to evaluate miR-21-5p expression in ESCC and adjacent tissue samples, followed by correlation analyses between Pg and LC3, and Pg and miR-21-5p. Results Pg infected KYSE70 cells and KYSE140 cells showed upregulation Atg7 protein and LC3-Ⅱ/LC3-Ⅰ protein but downregulation of RASA1 protein and p62 protein, enhanced cell proliferation, migration, and invasion as well as immunofluorescent spots of red, green, and yellow in mRFP-GFP-LC3-labeled ESCC cells. Pretreatment with CQ or siAtg7 abolished the above alterations induced by Pg. Consistently, pretreatment with miR-21-5p inhibitor, U0126, or RASA1 overexpression plasmid also blocked Pg-stimulated autophagy. In ESCC samples, Pg abundance was correlated with upregulation of miR-21-5p and LC3. Conclusion Pg promotes autophagy in esophageal squamous cell carcinomaviamiR-21-5p/RASA1/ERK signaling pathway.

abstract:
Objective To understand how warfarin inhibition is hindered by the vitamin K epoxide reductase(VKORC1) resistant mutations. Methods An electrophoretic mobility assay was conducted based on the principle that binding with warfarin resulted in changes in VKORC1 electrophoretic mobility. The activity of VKORC1 and the half maximal inhibitory concentration(IC50) of warfarin inhibiting VKORC1 were detected by ELISA. These experiments evaluated the binding ability of VKORC1 resistant mutants to warfarin. Results With warfarin bound, VKORC1 was protected from N-ethylmaleimide(NEM) modification and showed increased electrophoretic mobility, which was dependent on an unmodified cysteine on the flexible transmembrane helix 1(TM1) of VKORC1. Increasing the warfarin concentration could shift more VKORC1 towards the species with the fast mobility. Compared to the wildtype VKORC1, the fast mobility fraction became less or disappeared in warfarin-resistant mutants, indicating weakened binding of warfarin. In addition, VKORC1 mutants with the weaker electrophoretic mobility shift indicated the stronger warfarin resistant. Conclusion Weakened warfarin binding is the primary cause of warfarin resistance.

abstract:
Objective To explore the effect and mechanism of adenosine A2A receptor(A2AR) on lipopolysaccharide(LPS)-induced inflammatory injury of Caco-2 intestinal epithelial cells. Methods Caco-2 cells were divided into control group, LPS group(treated with 10 μg/ml LPS for 12 h), A2AR agonist(CGS21680) group(pretreated with 50 nmol/L CGS21680 for 10 min), CGS21680+LPS group(pretreated with 50 nmol/L CGS21680 for 10 min, treated with 10 μg/ml LPS for 12 h), cell viability was determined using CCK-8 assay, the secretion levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) and interleukin-6(IL-6) in cell supernatant of each group were determined using ELISA. mRNA expression levels ofTNF-α,IL-1β and IL-6in cells of each group were detected by real-time fluorescence quantitative PCR, the protein expression levels of microtubule associated light chain protein 3(LC3)-Ⅱ/LC3-Ⅰ and autophagy associated protein(Beclin1) in cells of each group were analyzed using Western blot analysis. Caco-2 cells were then divided into control group, LPS group(pretreated with 50 nmol/L CGS21680 for 10 min), CGS21680+LPS group(pretreated with 50 nmol/L CGS21680 for 10 min, treated with 10 μg/ml LPS for 12 h), CGS21680+LPS+Rapa group(pretreated with 50 nmol/L CGS21680 for 10 min, treated with 10 μg/ml LPS and 5 μmol/L Rapa for 12 h), cell viability was determined using CCK-8 assay, the secretion levels of TNF-α, IL-1β and IL-6 in cell supernatant of each group were determined using ELISA. Results Compared with the control group, the viability of Caco-2 cells in LPS group significantly decreased(P<0.05), the levels of TNF-α, IL-1β and IL-6 in supernatant significantly increased(P<0.05), the mRNA relative expressions ofTNF-α,IL-1β,IL-6in cells significantly increased(P<0.05), the LC3-Ⅱ/LC3-Ⅰ ratio and the relative expression of Beclin1 protein were significantly up-regulated(P<0.05). Compared with LPS group, the viability of Caco-2 cells in CGS21680+LPS group significantly increased(P<0.05), the levels of TNF-α, IL-1β and IL-6 in supernatant significantly decreased(P<0.05), the mRNA relative expression levels ofTNF-α,IL-1β,IL-6in cells were significantly down-regulated(P<0.05), and the ratio of LC3-Ⅱ/LC3-Ⅰ and the relative expression of Beclin1 protein were significantly down-regulated(P<0.05). In addition, compared with CGS21680+LPS group, the viability of Caco-2 cells in CGS21680+LPS+Rapa group significantly decreased(P<0.05), and the levels of TNF-α, IL-1β and IL-6 in the supernatant also significantly increased(P<0.05). Conclusion A2AR agonist can reduce the inflammatory injury of Caco-2 intestinal epithelial cells induced by LPS and improve cell viability, which may be related to its inhibition of autophagy.

abstract:
Objective To investigate the risk factors of short-term adverse prognosis and the predictive value of Charlson comorbidities index(CCI) in patients with central pulmonary embolism(PE). Methods 115 cases of central PE patients were retrospectively analyzed. According to the adverse prognosis during hospitalization, the subjects were divided into adverse event group and no adverse event group. The clinical characteristics of the adverse event group were analyzed. Multivariate Logistic regression analysis was performed for statistically significant indicators. Results The most common clinical symptoms of central PE patients were chest distress or dyspnea(77.4%), followed by cough(35.7%), chest pain(28.7%), syncope(9.6%) and hemoptysis(7.8%). There were no statistically significant differences in gender, smoking history, drinking history, symptoms and signs between the two groups. In univariate analysis, CCI, grouping score of thrombus location, white blood cell count, neutrophil count and urea nitrogen were associated with adverse events in central PE patients, with statistical significance(P<0.05). After Logistic regression multivariate analysis, increased neutrophil count(OR=1.494, 95%CI: 1.073-2.080,P=0.017) was an independent risk factor(P<0.05). The CCI in the group with adverse events was higher than that in the group without adverse events(P=0.004). Multivariate analysis showed that increased CCI(OR=1.342, 95%CI: 1.022-1.763,P=0.034) was an independent risk factor, and the risk of adverse events increased by 34.2% for every one-point increase in CCI. The thrombus location score of the group with adverse events was significantly higher than that of the group without adverse events(OR=2.586, 95%CI: 1.366-4.896,P=0.004), and the risk of adverse events increased 1.586 times with each increase of thrombus location score. Conclusion Increased neutrophil count, CCI, and thrombus location score are associated with poor short-term prognosis in central PE patients.

abstract:
Objective To investigate the ocular biological parameters of children with idiopathic short stature(ISS) and compare them with those of children with growth hormone deficiency(GHD) and normal children, and to explore the characteristics of ocular biological parameters in this group, so as to provide a reference for the screening of visual acuity and the safety of growth hormone therapy in children with ISS. Methods A total of 15 children aged 5-14 years old with ISS were selected as the observation group, 32 children with GHD were selected as the control group, and 47 children of normal height who underwent routine visual acuity screening were selected as normal controls. The ocular biological parameters of children with ISS were studied. The differences of vision-related parameters between the above three groups were compared. The influencing factors affecting the visual development of children with ISS were analyzed. Results The axial ratio of ISS was significantly higher than that of the GHD group and normal children, and the intraocular pressure of the ISS group was significantly higher than that of the GHD group and normal children. There was no significant difference in axial length between the ISS group and the GHD group, as well as normal children(P>0.05), but the axial length of the GHD group was significantly shorter than that of normal children. The corneal curvature of ISS was significantly greater than that of normal children. The axial rate ratio of the ISS group was positively correlated with the peak value and corneal curvature of growth hormone provocation test(β=1.052,P<0.05; β=0.004,P<0.05). Conclusion Children with ISS may have high intraocular pressure and high risk of myopia. Higher peak results of growth hormone provocation test and large corneal curvature may be the risk factors for myopia.

abstract:
Objective To understand the carrying situation and common variation of pathogenic genes of single gene hereditary disease in childbearing age population in Anhui province, to explore the establishment of clinical application network and referral model of carrier screening in Anhui province, and to explore the application value of expansible carrier screening(expanded carrier screening, ECS) in clinic. Methods Samples were collected from 604 individuals of childbearing age, all exhibiting a normal phenotype and a family history of inherited disease. These samples were obtained during the first trimester or early stages of pregnancy(≤13+6 weeks). Based on high-throughput sequencing and special PCR analysis techniques, pathogenic variants associated with 220 diseases were detected, and related genes were detected in the spouses of positive carriers. Results As of May 16, 2023, 604 tested samples had been collected, and 340 carriers of the target disease had been detected; The positive rate of pathogenic variation detection was 56.29%; A total of 499 pathogenic variants were detected, with each tested individual carrying 0-5 variants; 216 cases, accounting for 35.76%, carried a single gene recessive disease pathogenic variation, which was the most common. There were 95 cases carrying two types of single gene recessive genetic disease pathogenic variation, accounting for 15.73%. As of now, 302 couples have been reported, and a total of 7 high-risk couples have been found through screening, with a high-risk rate of 2.32%. There are a total of 5 pairs with autosomal recessive genetic pattern(both spouses carry the same pathogenic gene), and 2 pairs with X-linked genetic pattern(the female carries the X-linked pathogenic gene). Conclusion In this study, we obtained the overall carrier and clinical application of target diseases as well as the carrier rates of causative genes of common single-gene genetic diseases in 604 subjects who underwent ECS testing, which could provide scientific guidance for the establishment of a clinical application network and referral model for carrier screening in Anhui Province.

abstract:
Objective To explore the changes in posterior tibial slope after tibial plateau fracture surgery and its relationship with knee joint function. Methods The cases of 83 patients who underwent tibial plateau fracture surgery were collected. The posterior tibial slope(PTS) of the patient's unaffected side, the affected side after surgery and the affected side at the last follow-up were measured. He knee joint score data of Lysholm, HSS and Rasmussen were evaluated and recorded, and the patients were divided into PTS<9° group and PTS≥9° group according to the PTS angle after operation. The mean of three kinds of knee joint function scores in the two groups were statistically analyzed, and the mean of PTS after operation and final follow-up was also statistically processed. The difference of PTS between the last follow-up and the healthy side was calculated, and the correlation between the difference and the knee joint function data at the last follow-up was analyzed. Results There was no significant difference in general conditions between the two groups of patients. The Lysholm, HSS, and Rasmussen score data of knee joints in PTS<9° group were significantly higher than those in the PTS ≥9° group(P<0.05). There was a significant difference in postoperative PTS between the two groups of patients compared to the last follow-up PTS(P<0.05). The difference between the last follow-up and the PTS of the unaffected side(b) was correlated with Lysholm, HSS and Rasmussen knee score data(P<0.05). Conclusion Postoperative rehabilitation exercise leads to changes in the angle of PTS. The knee joint function of patients in PTS<9° group is better than that of patients in group PTS ≥9° after surgery. The difference between the last follow-up PTS and the healthy side PTS is correlated with knee joint function. Therefore, it is necessary to try to restore the anatomical alignment of the plateau fracture during surgery or control postoperative PTS<9°.

abstract:
Objective To investigate the difference of facial soft tissue changes in patients with different vertical bone facial types after orthodontic treatment.Methods A total of 137 female patients with class Ⅱ malocclusion aged 18 to 30 years old were selected for retrospective analysis using facial soft tissue 3D model data.According to the mandibular plane angle(FH-MP) angle,they were divided into high angle group,average angle group and low angle group.The EinScan Pro 2X 2020 handheld high-precision 3D scanner was used to capture facial soft tissue images of patients before treatment(TO) and at 6 months during treatment(T1) and after treatment(T2).The patients' facial images were overlapped using reverse engineering software Geomagic Wrap 2021,and the differences within and between groups were statistically analyzed using SPSS 26.0 statistical software.Results Before and after orthodontic treatment,the average overall facial changes in the high angle group were(-3.25±0.22)mm,in the average angle group was(-3.28±0.30) mm,and in the average low angle group was(-3.69 ±0.36) mm.Compared with the other two groups,the changes in the low angle group decreased more,and the difference was statistically significant(P<0.05).The mandibular angle area and temporal area decreased the most in the low angle group,which were(-2.78±0.18) mm and(-2.27±0.35) mm,respectively,and the difference was statistically significant compared with the other two groups(P<0.05),while there was no statistically significant difference among the other groups(P>0.05).Conclusion The whole face and all facial regions of the three groups had some negative changes,but the collapse in the mandibular angle area and the temporal muscle area of the low angle group was more obvious than that of the other two groups.

abstract:
Objective To analyze the relationship between microorganisms and endodontic disease by using 16 S rDNA sequencing to compare the composition of the microbial community in the root canals of teeth with pulpitis and apical periodontitis.Methods Clinical samples were collected from teeth requiring root canal treatment.The total DNA of the bacteria in the samples and the gene fragments of the V3-V4 highly variable region on the 16S rDNA fragments were amplified through PCR.After sequencing by NovaSeq,statistical and bioinformatic analysis,including phylogenetic analysis,diversity analysis and analysis of group differences,were performed.Results In total,6 teeth with pulpitis and 7 teeth with apical periodontitis were collected,and a total of 8 510 OTUs were obtained after next-generation sequencing,and the analysis of bacterial diversity showed that the difference between pulpitis and apical periodontitis in terms of the composition of the bacterial flora was statistically significant(P<0.05).In particular,the relative abundance of Proteobacteria,Acidobacteriota and Actinobacteriota phylum was significantly higher in the roots of teeth affected by pulpitis than apical periodontitis.The relative abundance of Bacteroidota phylum and Synergistota phylum was significantly higher in the root canals of teeth with apical periodontitis.Conclusion There is a complex diversity of infecting microorganisms in the root canals of teeth affected by endodontic diseases.The microbial communities in the infected root canals of pulpitis and apical periodontitis show some differences,and changes in the microbial composition of the root canals may be associated with the development of endodontic diseases.

abstract:
The cranial suture complex is made up of fibrous tissue with mesenchyme, an osteogenic front, the dura mater beneath, and an overlying periosteum. The dura mater is a layer of fibrous connective tissue that protects the brain and spinal cord, containing blood and lymphatic vessels. It is important for the progression of cranial osteogenesis during early development, as well as the cranial morphology and the state of the cranial suture later in life. Cranial suture mesenchymal stem cells are primarily found in the cranial suture and can proliferate, differentiate into osteoblasts, generate cranial bone, and aid in the repair of cranial bone after injury. Understanding how cranial suture mesenchymal stem cells interact with the dura mater is critical for cranial growth and development, as well as the treatment of cranial diseases. This article reviews the roles of dura mater and cranial suture mesenchymal stem cells in cranial bone formation.

abstract:
Epithelial splicing regulatory protein 1(ESRP1) is an epithelial cell-specific splicing factor that involved in selective splicing and translation of multiple genes. ESRP1 regulates signaling pathways that affect cell proliferation and tumor growthviamediating target genes and regulating cyclic RNA cyclization and biogenesis in a variety of cancers, including head and neck squamous cell carcinoma, carcinoma of colon, and prostate cancer. Through integrating and analyzing the link between ESRP1 and related cancers, provide new ideas for the treatment of tumor cells in which ESRP1 as a key factor.