Objective To investigate the expression of FBXO43in various cancers and its relationship with immune cell infiltration and prognosis using bioinformatics.Methods Gene expression data for 33 cancers were obtained from The Cancer Genome Atlas(TCGA)and Genotype-Tissue Expression(GTEx)databases to analyze FBXO43expression.Clinical and survival data were sourced from the TCGA database.Gene Expression Profiling Interactive Analysis(GEPIA)was used to assess the correlation between FBXO43expression and the clinical stage.Kaplan-Meier survival analysis was used to evaluate the relationship between FBXO43expression and prognosis.R lan-guage was used to analyze the associations between FBXO43expression and clinical stages,immune cell infiltration,immune checkpoint genes,tumor mutation burden(TMB),microsatellite instability(MSI),and mismatch repair(MMR)genes.The potential biological mech-anisms of FBXO43were explored using gene set enrichment analysis(GSEA).Moreover,qRT-PCR was performed to measure FBXO43 expression in normal gastric mucosal cells(GES-1),gastric cancer cells(HGC-27,MGC-803,and MKN-45),normal liver cells(LO-2),and liver cancer cells(SMMC-7721,HEPG2,HuH7,and MHCC97-H).Results Combined TCGA and GTEx database statistics revealed higher FBXO43expression in 26 types of cancer tissues,including adrenocortical carcinoma(ACC),bladder urothelial carcinoma,breast invasive carcinoma,cervical squamous cell carcinoma and endocervical adenocarcinoma,cholangiocarcinoma,colon adenocarcinoma,and diffuse large B-cell lymphoma,than in normal tissues(P<0.05).However,FBXO43expression was lower in three types of cancer tissues:kidney chromophobe(KICH),testicular germ cell tumor(TGCT),and thyroid carcinoma(P<0.05).GEPIA data analysis showed that FBXO43expression positively correlated with the clinical stages of ACC,KICH,kidney renal clear cell carcinoma(KIRC),and kidney renal papillary cell carcinoma(KIRP)and negatively correlated with the clinical stages of ovarian serous cystadenocarcinoma(OV)and TGCT(P<0.05).Kaplan-Meier survival analysis indicated that abnormal FBXO43expression was associated with the prognosis of various cancers(P<0.05).Specifically,high FBXO43expression was a risk factor for ACC,KICH,KIRC,KIRP,low-grade glioma,liver hepato-cellular carcinoma,mesothelioma,and sarcoma,but protective in thymoma(THYM).The XCELL algorithm found that FBXO43expres-sion was negatively correlated with immune scores in nine types of cancer tissues,including glioblastoma multiforme,KIRP,acute myeloid leukemia,lung squamous cell carcinoma,and OV,and closely related to immune cell infiltration levels in 24 types of cancer tissues,espe-cially showing a significant negative correlation with most immune cells in SARC(P<0.01).Correlation analysis revealed a significant association between FBXO43and TMB,MSI,and MMR in all cancers.GSEA analysis indicated that FBXO43is involved in the cell cycle and immune-related functions in various tumors.qRT-PCR results showed that FBXO43expression was upregulated in liver cancer cells and downregulated in gastric cancer cells(all P<0.05).Conclusion Abnormal FBXO43expression is closely associated with the occur-rence and progression of multiple cancers.Thus,FBXO43may serve as a novel marker of immune cell infiltration and prognosis,thereby offering new directions for targeted cancer therapy.

Objective To explore the correlations between serum soluble growth stimulation gene 2 protein (sST2), brain natriuretic peptide (BNP) and the degree of carotid atherosclerosis in patients with acute ischemic stroke. Methods A total of 205 patients with acute ischemic stroke were selected as study group, and were divided into mild group with 100 cases, moderate group with 70 cases, and severe group with 35 cases according to the severity of carotid atherosclerosis. Another 150 healthy people who received physical examination during the same period were selected as control group. Serum levels of sST2 and BNP were compared among all groups. The correlations of sST2 and BNP levels with the degree of carotid atherosclerosis were analyzed. The influencing factors of different degree of carotid atherosclerosis in patients with acute ischemic stroke were analyzed. The predictive value of sST2 combined with BNP on the degree of carotid atherosclerosis in patients with acute ischemic stroke was analyzed. Results The serum sST2 level of the study group was significantly lower than that of the control group, and the serum BNP level was significantly higher than that of the control group (P < 0.05). The sST2 levels in moderate and severe groups were significantly lower, BNP levels were significantly higher than that in the mild group (P < 0.05). BNP level in the severe group was significantly higher, sST2 level was significantly lower than that in the moderate group (P < 0.05). The degree of carotid atherosclerosis in patients with acute ischemic stroke was correlated with age, history of hypertension, history of smoking, history of diabetes, levels of total cholesterol (TC), C-reactive protein (CRP), homocysteine (Hcy) and low density lipoprotein (LDL) (P < 0.05). Age, hypertension history, smoking history, diabetes history, TC, CRP, Hcy, LDL, BNP and sST2 were the influencing factors for degree of carotid atherosclerosis (P < 0.05). Serum BNP level was positively correlated with the degree of carotid atherosclerosis, while sST2 level was negatively correlated with the degree of carotid atherosclerosis (P < 0.001). The area under the curve (AUC) for the combined prediction of sST2 and BNP was significantly greater than that for the individual prediction of sST2 and BNP (P < 0.001). Conclusion Clinical observation of the changes in serum levels of sST2 and BNP in patients with acute ischemic stroke can effectively assess the progression of the disease, which is beneficial for improving the severity of carotid atherosclerosis in patients. The combined assessment of sST2 and BNP shows good diagnostic performance for evaluating the severity of carotid atherosclerosis in patients with acute ischemic stroke.

Marsdenia tenacissima injection, a standard Marsdenia tenacissima extract (MTE), has been approved as an adjuvant therapeutic agent for various cancers. Our previous study showed that MTE inhibited the proliferation and metastasis of prostate cancer (PCa) cells. However, the underlying mechanisms and active ingredients of MTE against PCa were not completely understood. This study revealed that MTE induced significant decreases in cell viability and clonal growth in PCa cells. In addition, MTE induced the apoptosis of DU145 cells by reducing the mitochondrial membrane potential and increasing the expression of Cleaved Caspase 3/7, Cyt c, and Bax. In vivo, DU145 xenografted NOD-SCID mice treated with MTE showed significantly decreased tumor size. TUNEL staining and Western blot confirmed the pro-apoptotic effects of MTE. Network pharmacology analysis collected 196 ingredients of MTE linked to 655 potential targets, and 709 PCa-associated targets were retrieved, from which 149 overlapped targets were screened out. Pathway enrichment analysis showed that the HIF-1, PI3K-AKT, and ErbB signaling pathways were closely related to tumor apoptosis. Western blot results confirmed that MTE increased the expression of p-AKT^Ser473 and p-GSK3β^Ser9, and decreased the expression of p-STAT3^Tyr705in vitro and in vivo. A total of 13 compounds in MTE were identified by HPLC-CAD-QTOF-MS/MS and UPLC-QTOF-MS/MS. Molecular docking analysis indicated that six compounds may interact with AKT, GSK3β, and STAT3. In conclusion, MTE induces the endogenous mitochondrial apoptosis of PCa by regulating the AKT/GSK3β/STAT3 signaling axis, resulting in inhibition of PCa growth in vitro and in vivo.
Mice ; Animals ; Male ; Humans ; Mice, Inbred NOD ; Mice, SCID ; Marsdenia ; Proto-Oncogene Proteins c-akt ; Glycogen Synthase Kinase 3 beta ; Molecular Docking Simulation ; Phosphatidylinositol 3-Kinases ; Tandem Mass Spectrometry ; Prostatic Neoplasms ; Apoptosis ; STAT3 Transcription Factor

Objective: To investigate the epidemiological characteristics of pulmonary tuberculosis ( PTB ) incidence and mortality in Zhejiang Province from 2016 to 2020, so as to provide the evidence for formulating the PTB control strategy. Methods: The data regarding the PTB patients in Zhejiang Province from 2016 to 2020 were collected from the Tuberculosis Management Information System of the Chinese Disease Control and Prevention Information System. The reported cases and deaths of PTB cases were descriptively analyzed by time, population and regions. Results: A total of 129 985 PTB cases were reported in Zhejiang Province from 2016 to 2020, and the overall reported incidence rate of PTB was 45.81/105, with a tendency toward a decline in the reported incidence ( χ2trend=298.899, P<0.001 ) and an annual decline rate of 3.72%. The prevalence rates of rifampicin resistance, positive etiological tests, negative etiological tests and no etiological results were 1.51%, 47.17%, 46.67% and 4.65%, respectively. A total of 546 PTB deaths occurred in Zhejiang Province from 2016 to 2020, and the reported mortality of PTB was 0.19/105, with a tendency towards a decline ( χ2trend=10.818, P<0.001) and an annual decline rate of 11.99%.The prevalence rates of rifampicin resistance, positive etiological tests, negative etiological tests and no etiological results were 1.83%, 82.97%, 13.37% and 1.83%, respectively. The highest reported incidence ( 92.18/105 ) and mortality ( 1.19/105 ) of PTB was found in patients aged 65 years and older, and farmer was the predominant occupation of PTB cases (60 774 cases, 46.75%) and dead PTB cases ( 345 cases, 63.19% ). In addition, the top three cities with the highest incidence of PTB included Quzhou City ( 66.43/105 ), Jinhua City ( 64.24/105 ) and Lishui City ( 53.91/105 ), and the top three cities with the highest mortality of PTB included Quzhou City ( 0.67/105 ), Lishui City ( 0.27/105 ) and Jiaxing City ( 0.26/105 ). Conclusions Both the reported incidence and mortality of PTB appeared a tendency towards a decline in Zhejiang Province from 2016 to 2020; however, the proportion of positive etiological tests increased. Higher attention should be paid to the elderly aged 65 years and older and and farmers, and active screening and health education are recommended; in addition, PTB control requires to be intensified in highly prevalent regions, including Quzhou City, Jinhua City and Lishui City.

Objective: To analyze the epidemiological characteristics of pulmonary tuberculosis cases at ages of 60 years and older in Quzhou City, Zhejiang Province, so as to provide the evidence for formulating the pulmonary tuberculosis control strategy among the elderly. Methods: The data pertaining to pulmonary tuberculosis cases at ages of 60 years and older in Quzhou City from 2010 to 2020 were collected through the Tuberculosis Management Information System of the China Disease Prevention and Control Information System. The trends for incidence, population distribution, diagnosis and treatment of pulmonary tuberculosis cases aged 60 years and older were analyzed using a descriptive epidemiological method. Results: Totally 8 754 pulmonary tuberculosis cases aged 60 years and older were reported in Quzhou City from 2010 to 2020, accounting for 47.65% of all pulmonary tuberculosis cases, and the number of pulmonary tuberculosis appeared a tendency towards a rise ( χ2trend=173.320, P<0.001 ), while the incidence of pulmonary tuberculosis showed a tendency towards a decline ( χ2trend=389.820, P<0.001 ), with an annual decline rate of 177.11/105. There were 4 307 smear-positive pulmonary tuberculosis cases, accounting for 58.71% of all smear-positive cases, and the number of smear-positive pulmonary tuberculosis cases showed a tendency towards a rise ( χ2trend=126.320, P<0.001 ), while the proportion of smear-positive cases showed a tendency towards a decline ( χ2trend=21.680, P<0.001 ), with an annual smear-positive rate of 87.14/105. The incidence of pulmonary tuberculosis was 256.94/105 among males and 91.43/105 among females at ages of 60 years and older ( χ2=20.903, P<0.001 ). The highest incidence of pulmonary tuberculosis was seen in patients aged 80 to 84 years ( 235.17/105 ), and farmers were the predominant occupation ( 7 171 cases, 81.92% ), while the highest number of cases was reported in Quzhou City (8 676 cases, 99.11%). There were 7 752 treatment-naïve cases ( 88.55% ), while 5 830 cases with delay in seeking healthcare services, and the proportion of delay in seeking healthcare services showed a tendency towards a rise from 2010 to 2020 ( χ2trend=4.853, P=0.028 ), with an annual mean delay rate of was 66.60%. Conclusion The incidence of pulmonary tuberculosis appeared a tendency towards a decline among patients aged 60 years and older in Quzhou City from 2010 to 2020, and the elderly aged 80 years and older and farmers should be paid more attention.

Objective:To investigate the effect of sialic acid combined with immunoglobulin-like lectin 15 (SIGLEC-15) on laryngeal squamous cell carcinoma (LSCC) and underlying mechanisms.Methods:The Cancer Genome Atlas (TCGA), Cancer Cell Line Encyclopedia (CCLE) and Gene Expression Profiling Interactive Analysis (GEPIA2) databases were used for bioinformatics analysis. Cell Counting Kit-8 (CCK8), flow cytometry, and Transwell method were used respectively to detect proliferation, apoptosis, cell cycle, metastasis and invasion behaviors of the cells. Gene chip method was used for detecting up-regulated and down-regulated genes and performing enrichment analysis of differential genes. Western Blotting (WB) and Quantitative Real-time PCR (qPCR) were used to detect the expressions of proteins. Tumor formation experiments in nude mice were used to detect the effect of SIGLEC-15 on the growth of transplanted tumors. Wilcoxon rank sum test, t-test and Log-rank test were used for statistical analysis. Results:SIGLEC-15 was highly expressed in laryngeal squamous cell carcinoma and closely related to life in being. TU686 SIGLEC-15-with low expression of SIGLEC-15 was constructed. Compared to TU686 SIGLEC-15+, TU686 SIGLEC-15-showed significantly reduced activities of proliferation (48 h: 1.32±0.23 vs. 2.56±0.37, t=6.59, P<0.05), migration (1 036.52±51.22 vs. 1 819.62±180.24, t=7.22, P<0.05) and invasion (469.21±112.25 vs. 961.45±102.03, t=7.85, P<0.05); early increased apoptosis ((23.27±1.12)% vs. (5.64±1.61)%, t=11.32, P<0.05); blocked cell cycle at G0/G1 ((59.32±3.65)% vs. (35.46±3.57)%, t=9.85, P<0.05); the knockdown of SIGLEC-15 resulted in up-regulation of 864 genes, down-regulation of 357 genes, with significant changes in molecules of cell cycle, apoptosis and JAK/STAT signal pathways, and the expressions of p-JAK2, p-STAT3, Caspase-3, Bad, Bcl-2, and Cyclin d1 proteins. Tumor formation experiments in nude mice showed that at 8 weeks after the tumors were implanted, the growth transplanted tumors of TU686 SIGLEC-15-cell group was slower than that of TU686 SIGLEC-15+cell group, with significant difference in the mean tumor weights between two groups ((0.382±0.054) g vs. (1.277±0.126) g, t=8.44, P<0.05), while the expression of SIGLEC-15 was lower in the transplanted tumors of SIGLEC-15 knockdown group compared to control group, with significant difference(11.29±2.17 vs. 36.25±7.56, t=9.28, P<0.05). Conclusion:SIGLEC-15 is highly expressed in LSCC and can promote the progression of LSCC through the JAK2-STAT3 pathway.

Objective:To observe the presence or absence of necroptosis in PC12 cells after radiation injury, and to detect the expression of receptor-interacting protein 3(RIP3) and evaluate its regulatory effect on necroptosis.Methods:PC12 cells were treated with different doses of irradiation and their necroptosis was detected by lactate dehydrogenase (LDH) release at different time points. After pretreatment with necroptosis inhibitor Necrostatin-1(Nec-1), the changes of cell necroptosis were detected by LDH. The expression level of RIP3 after irradiation intervention was detected by Western blot (WB). After pretreatment with the RIP3-specific inhibitor GSK′872, the changes of cell necroptosis were detected by LDH. The best transfection sequence of RIP3 knockout was screened by WB. The cells were divided into the control group, irradiation group, solvent control group, no-load control group and pretreatment group. WB, immunofluorescence staining, MTT, LDH and Annex V-fluorescein Isothiocyanate/Propidium Iodide (AnnexV-FITC/PI) flow cytometry were used for detection and analysis.Results:After 4 Gy irradiation, the degree of cell necrosis was the highest after 3 hours of culture, and the expression level of RIP3 protein was up-regulated. The cell necrosis was decreased after Nec-1, GSK′872 and RIP3 gene knockdown pretreatment.Conclusions:The radiation injury of 4 Gy can induce the necroptosis of PC12 cells, and the most significant effect can be observed when cultured for 3 hours after irradiation. RIP3 is involved in the process of necroptosis of PC12 cells induced by radiation injury, and plays a pivotal positive regulatory role.

Objective To explore the clinical feasibility of predicting synchronous liver metastases based on MRI radiomics nomogram based on T2WI in rectal cancer. Methods The imaging and clinical data of 261 patients with primary rectal cancer admitted to Zhejiang People′s Hospital from April 2012 to May 2018 were retrospectively analyzed. 101 patients were accompanied by synchronous liver metastasis All cases were divided into training group (n=182) and verification group (n=79). T2WI image of each patient was selected to extract texture features by AK analysis software of GE company. A radiomics signature was constructed after reduction of dimension in training group by the least absolute shrinkage and selection operator (LASSO). Univariate logistic regression was used to select for independent clinical risk factors and multivariate logistic regression along with imaging omics tags were used to construct predictive models and nomogram. ROC was used to assess the accuracy of the nomogram in the training group and to verify them by the validation group. Finally, the clinical efficacy of each patient′s synchronized liver metastasis risk factor was calculated based on the nomogram. Results A total of 328 texture features were extracted from the T2WI. Seven most valuable features were selected after reducing the dimension by LASSO algorithm, including 3 co-occurrence matrices (GLCM) and 4 run-length matrices(RLM). Tumor staging and radiomic signatures were included in the Multifactor logistic regression to build the prediction model and nomogram. The accuracy of predicting SRLM was 0.862 and 0.844 in the training and the verification group, respectively. To evaluate the accuracy of the nomogram, radiomics signature and the tumor staging in all cases were 0.857, 0.832 and 0.663, respectively. There was no significant difference in the number of SRLM cases between the high risk group and the low risk group based on nomogram (P>0.05). Conclusion The radiomics nomogram based on T2WI can be used as a quantitative tool to predict synchronous liver metastases of rectal cancer.

OBJECTIVETo evaluate the difference in the clinical therapeutic effects on anal fissure at Ⅰ and Ⅱ stages between the acupoint catgut embedding therapy and western medication.

METHODSSixty patients of anal fissure at Ⅰ and Ⅱ stages were randomized into an embedding therapy group and a western medication group, 30 cases in each one. In the embedding therapy group, the acupoint catgut embedding therapy was applied at bilateral Tianshu (ST 25), Changqiang (GV 1), bilateral Chengshan (BL 57) and Tigangxue (Extra), once a week. In the western medication group, the external inunctum on the wound was given with 0.2% nitroglycerin ointment, once every morning and evening a day. The treatment lasted for 4 weeks continuously in the two groups. The follow-up visit was done for 3 months after treatment. The visual analogue scale (VAS) and anal pain duration were observed and recorded before treatment and on the 3rd day and the 7th day of treatment separately. The clinical therapeutic effects were compared between the two groups.

RESULTSAfter treatment, on the 3rd day and the 7th day of treatment, VAS score and anal pain duration were all reduced significantly as compared with those before treatment in the patients of the two groups (all<0.01). The differences in the embedding therapy gruop were better than those in the western medication group before and after treatment (<0.01,<0.05). In the 2nd and 4th weeks after treatment, the clinical therapeutic effects in the embedding therapy group were better than those in the western medication group (both<0.05). In 3-month follow-up, the recurrent case in the embedding therapy group was one, and the recurrent case in the western medication group was six.

CONCLUSIONSThe acupoint catgut embedding therapy is safe and effective in the treatment of anal fissure at Ⅰ and Ⅱ stages and its recurrent case is lower as compared with the treatment of western medication.

BACKGROUND:The etiology of anterior cruciate ligament injury remains unclear yet, and some researchers have pointed that interior and exterior factors both contribute to anterior cruciate ligament injury;additionally, the genetic factor interior factors stand out. Collagen genes COL1A1, COL5A1, and COL12A1 are reported to be associated with anterior cruciate ligament injury in Caucasian populations. OBJECTIVE:To investigate the association of polymorphisms of COL1A1, COL5A1 and COL12A1 genes with anterior cruciate ligament injury in Chinese Han population . METHODS:105 patients with anterior cruciate ligament injury were enrolled and 110 patients without history of anterior cruciate ligament injury were as controlls. The first intron rs1800012 in COL1A1, rs127722 and rs13946 in the 3'-UTR region of COL5A1 gene, rs970547 and rs240736 in the 65 and 29 regions of COL12A1 extron were detected and classified by restriction fragment length polymorphism and genetic sequencing technology. RESULTS AND CONCLUSION:rs1800012, rs12722 and rs13946 genotypes, phenotypes and haplotypes in COL1A1 and COL5A1 genes showed no significant differences between groups. rs970547 and rs240736 genotypes as well as phenotypes and haplotypes in COL12A1 also showed no significant differences between groups. However, there was a significant difference in rs970547 gene frequence in male patients between groups. In conclusion, the Sp1 binding site of COL1A1 rs1800012 is not the susceptibility locus of anterior cruciate ligament injury in Chinese Han population. COL5A1 genes rs12722 and rs13946 in COL5A1 are not closely related to anterior cruciate ligament injury. COL12A1 rs970547 and rs240736 have a certain association with anterior cruciate ligament injury in Chinese men. Male individuals with COL12A1 rs970547 A allelicgene and AA genotype are likely to be susceptible to anterior cruciate ligament injury in Chinese Han population.