Objective To study the changes in miRNAs expression in the exosomes of human umbilical vein endothelial cells(HUVECs) after 60 Co γ-rays expose using microRNA(miRNA) chips and bioinformatics techniques so as to provide new clues to the mechanism of radiation-induced vascular tissue injury and its bystander effects.Methods HUVECs exosomes were collected in the control and 4 Gy irradiated cells by ultra-high-speed centrifugation,and further confirmed using transmission electron microscopy (TEM) and Western blotting of exosomes biomarkers.miRNA microarray was used to analyze miRNA expression profiles of exosomes and cells.Also,real-time quantitative PCR(qRT-PCR) was used to verify differentially expressed miRNAs,and the miRDB and TargetScan were performed to predict the target genes of the differentially expressed miRNAs.Bioinformatics analysis was performed using DAVID,KEGG and other online tools.Results Compared with the control exosomes from non-irradiated HUVECs,miRNA microarray analysis revealed that 5 up-regulated,and 13 down-regulated miRNAs were identified in the exosomes from HUVECs at 0.5 h after 4 Gy-irradiation,and 16 up-regulated and 5 down-regulated miRNAs at 2 h after 4 Gy-irradiation.Moreover,38 and 85 miRNAs were differentially expressed respectively in the HUVECs at 0.5 h and 2 h after radiation.The difference was statistically significant(P<0.01).The results of bioinformatics showed that these miRNAs might exert the radiation-induced bystander effect (RIBE) by regulating MAPK signal pathways,RAS and PI3K-Akt signal pathways.Conclusion The ionizing radiation injury significantly alters the components and expression levels of exosomal miRNAs,which play important roles in regulating the signal pathways in response to radiation.

Objective To investigate the correlation of P-gP,bel-2,STAT3,IL-6,IL-10 and the resistance lymphoma cells to chemotherapeutic agents.Methods There were 28 patients,among them were divided into three groups:refractory lymphoma,lymphoma which is sensitive to agents and lymphodenitis.P-glycoprotein(P-gp)on lymphoma cell membrane,STAT3,bcl-2,IL-6 and IL-10 level into lymphonla cell were detected using FCM,and the correlation between them and chemotherapy efficacy were analysed. Results In refractory lymphoma patients,P-gp and hcl-2 are significant higher than that of the group which is sensitive to agents(P:0.01,P=0.039),but STAT3,IL-6 and IL-10 were not significant different between these two groups(P>o.05).P-gp is significant higher in lymphoma than in lymphodenitis(P=0.01).STAT3 in lymphoma is significant lower than that of in lymphodenitis (P=0.04).The level bcl-2,IL-6 and IL-10 between lymphoma and lymphodenitis are not significant different(P>0.05).Conclusion The expression level of bel-2 and p-gP is correlated to resistance to the chemotherapeutic agents in lymphoma cells.STAT3 play a role in lymphoma cell signal transduction,but it is not certain in lymphoma cell muhidrug resistance.

Objective To investigate the correlation between XRCC1 R280H,XRCCl TSS+29C/T genetic polymorphisma and susceptibility to non-Hodgkin lymphoma (NHL). Methods The MassARRAY method was applied to detect the DNA repair gene XRCC1 genetic polymorphisms in 73 cases of NHL and 540 cases of normal healthy controls. Chi-square test was performed to calculate the adjusted odds ratios (OR) and 95% confidence intervals (CI). Results For XRCCl R280H genotypes, there was a significant difference between frequencies of the G and A among patients and controls (P=0.001). However, XRCCl TSS+29C/T genotypes had no statistical difference as for the T and C frequencies between patients and controls (P = 0.383). The frequency of XRCCI R280H with at least one A genotype was lower in the NHL cases than in controls, indicating a decreased risk for NHL development (OR=0.309, 95 % CI =0.168-0.567), comparing with GG genotype. In XRCC1 TSS+29C/T genotypes, the frequeney of TC and CC genotype was higher in NHL cases than in controls and associated with an increased risk of NHL development (P=0.472, OR =1.262, 95 % CI =0.669-2.379). Conclusion DNA repair XRCCl gene possesses significant correlation with NHL.

Objective To evaluate percutaneous intratumoral injection of chemotherapeutic agents lipiodol emulsion (CALE) for the treatment of primary liver cancer. Methods This study included 57 patients of hepatocellular carcinoma (n=49) and intrahepatic cholangiocarcinoma (n=8).53 were male and 4 were female,with a mean age of 48.02 years(range,19～70 years).In all,ninety CALE injections were assigned to 90 target areas within the lesions.Before the procedures,transcathetcr arterial chemoembolization (n=55) or infusion (n=2) was carried out in these patients.By arteriography,low blood supply of target areas was showed or it was concluded that superselective catheterization of supply arteries of lesions could not accomplished.Percutaneous intratumoral CALE injection was carried out under fluoroscopy or CT guidance.Therapeutic effect,side effect and complications were assessed based on clinical manifestation,laboratory examination and fluoroscopy or CT one week after procedure.Follow-up was carried out after 1,3,6 months and 1 year,and once every six months thereafter.Local recurrences were treated according to patients'will.Results 90 sessions of percutaneous injection were successfully performed on 57 patients,with 100% technique success rate.The volume of CALE iniected per session ranged 3.0-7.0 ml(mean,6.0 ml) in target size less than 3 cm,12.0-20 ml(mean,15 ml)in target size of 3-5 cm and 24-40 ml in target larger than 5 cm.Serum AFP was positive in 43 patients and decreased to normal in 14 patients(28%).54 lesions(60%)were with well distribution of the lipiodol-chemotherapy mixture on CT 1 month after procedure.Follow-up ranged from 2 months to six years(mean,16 months).The median survival time was 400 d.The cumulative survival rates at 200 d,600d was 85%and 30%,respectively.Complications included fever (n=22,24.4%),nausea and vomit(n=11,12.2%),and pain at the puncture site (n=17,18.9%).Conclusions Percutaneous intratumor CALE injection is safe and effective for the treatment of primary liver cancer.

Communication between doctors and patients is an important way to establish good doctorpatient relationship.It is crucial to master doctor-patient communication skills and arts for general practitioners who will extensively serve the people in community for life-long time after completion of residency training.To improve the communication skills and to enhance the clinical competency of general practitioners,we applied the Balint special group activities as core of doctor-patient communication course in residency training program.Through 3 years of practice,we found that the application of Balint group enriched the teaching contents and form of general practice residency training;improved doctor-patient communication skills,and enhanced the competency and professionalism of general practice trainees.

OBJECTIVE: To investigate the effect of danusertib (Danu), an inhibitor of Aurora kinase, on the proliferation, cell cycle, apoptosis, and autophagy of hepatocellular carcinoma HepG2 cells and explore the underlying mechanisms. METHODS: MTT assay was used to examine the effect of Danu on the viability of HepG2 cells to determine the IC50 of Danu. The effect of Danu on cell cycle distribution, apoptosis and autophagy were determined using flow cytometry. Western blotting was used to detect the expressions of the proteins related to cell cycle, apoptosis and autophagy. Chloroquine was used to suppress Danuinduced autophagy to test the apoptosis-inducing effect of Danu. RESULTS: Danu significantly inhibited the proliferation of HepG2 cells with IC of 39.4 μmol and 14.4 μmol at 24 h and 48 h, respectively. Danu caused cell cycle arrest in G/M phase in HepG2 cells and led to polyploidy accumulation via up-regulating the expressions of p53 and p21 and down-regulating the expressions of cyclin B1 and DC2. Danu also caused apoptosis of HepG2 cells through up-regulating the expressions of Bax, Puma, cleaved caspase-3, cleaved caspase-9, cleaved PARP and cytochrome C and down-regulating the expressions of Bcl-xl and Bcl-2. Danu induced autophagy via activating AMPK signaling and inhibiting PI3K/PTEN/AKT/mTOR axis, and inhibition of Danu-induced autophagy with chloroquine enhanced the pro-apoptotic effect of Danu. CONCLUSIONS Danu inhibits cell proliferation and induces cell cycle arrest in G/M phase, apoptosis and cytoprotective autophagy in HepG2 cells.
Apoptosis ; drug effects ; Autophagy ; drug effects ; Benzamides ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; Cell Proliferation ; drug effects ; Hep G2 Cells ; Humans ; Liver Neoplasms ; pathology ; Neoplasm Proteins ; metabolism ; Protein Kinase Inhibitors ; pharmacology ; Pyrazoles ; pharmacology

Objective:To investigate the diagnostic value of independent and combined subtests of the psychometric hepatic encephalopathy score (PHES) in mild hepatic encephalopathy(MHE) of patients with liver cirrhosis, so as to optimize the PHES.Methods:This was a prospective, multicenter and real-world study which was sponsored by the National Clinical Research Center of Infectious Diseases and the Portal Hypertension Consortium. Twenty-six hospitals from 13 provinces, autonomous regions and municipalities countrywide participated in this study, induding Tianjin Third Central Hospital, the Fourth People′s Hospital of Qinghai Province, the Second Affiliated Hospital of Baotou Medical College, the Third People′s Hospital of Taiyuan, the Fifth Medical Center of PLA General Hospital and so on. From October 2021 to February 2022, outpatients and hospitalized patients with liver cirrhosis and no obvious hepatic encephalopathy were consecutively enrolled. All patients received 5 PHES subjects in the same order: number connection test(NCT)-A, NCT-B, digit symbol test(DST), line tracing test(LTT) and serial dotting test(SDT), and the scores were calculated. The total score of PHES <-4 was taken as the cut-off value for diagnosing MHE. Compare the differences in each subtest between MHE group and non-MHE group. Receiver operating characteristic curve(ROC) and area under the curve(AUC) was performed to assess the diagnostic value of independent and combined subtests in MHE. Mann-Whitney U test and DeLong test were used for statistical analysis. Results:A total of 581 patients with liver cirrhosis were enrolled, 457 were diagnosed as MHE, and the incidence of MHE was 78.7%. The results of NCT-A, NCT-B, SDT, LTT, DST of MHE group were 60.00 s(47.01 s, 88.00 s), 90.45 s(69.32 s, 125.35 s), 74.00 s(57.65 s, 96.60 s), 74.72(60.00, 98.61) and 27.00(20.00, 36.00), respectively. Compared those of non-MHE group(34.00 s(29.15 s, 44.48 s), 50.00 s(40.98 s, 60.77 s), 50.00 s(41.07 s, 63.03 s), 46.23(38.55, 59.42) and 42.00(34.00, 50.75)), the differences were statistically significant( Z=12.37, 12.98, 9.83, 11.56, 10.66; all P<0.001). The AUC(95% confidence interval(95% CI)) of subtests of PHES NCT-B, NCT-A, LTT, DST and SDT alone in MHE diagnosis were 0.880(0.849 to 0.910), 0.862(0.828 to 0.896), 0.838(0.799 to 0.877), 0.812(0.772 to 0.851) and 0.788(0.743 to 0.832), respectively. The combination of 2 PHES subtests significantly increased the diagnostic efficacy. Among them the diagnostic efficacy of the combination of NCT-B and LTT was the best, the AUC(95% CI) was 0.924(0.902 to 0.947), the specificity was 91.9% and the sensitivity was 79.2%, which was better than a single PHES subtest (NCT-A, NCT-B, SDT, LTT and DST) and the combination of NCT-A and DST(AUC was 0.879, 95% CI0.847 to 0.910) which was recommended by guidelines on the management of hepatic encephalopathy in cirrhosis, the differences were statistically significant ( Z=3.78, 3.83, 5.57, 5.51, 5.38, 2.93; all P<0.01). Furthermore, compared between the combination of NCT-B and LTT and the combination of 3 subests of PHES, only the diagnostic efficacy of combination of NCT-B, LTT and SDT (AUC was 0.936, 95% CI 0.916 to 0.956) was better than that of the combination of NCT-B and LTT, the difference was statistically significant( Z=2.32, P=0.020). Conclusion:Based on the diagnostic efficacy and clinical feasibility of PHES subtests and their combinations, the combination of NCT-B and LTT is recommended for the diagnosis of MHE.