Objective To study the expression of TFIIB-related factor 2 (BRF2) in hepatocellular carcinoma(HCC) and to determine its clinical significance.Methods 80 HCC patients who received ‘curative' hepatectomy at the Qilu Hospital were studied.Immunohistochemistry analysis was performed to examine the expression of BRF2 and CD34 (microvessel density) in tumor tissues,matched paraneoplastic tissues,and normal liver tissues.Statistical methods were performed to analyse the relationships of expressions of BRF2 and CD34 with clinicopathologic factors.Results BRF2 protein was expressed at a high rate of 61.3％ in HCC tumor,which was significantly higher than that in normal liver tissues and matched paraneoplastic tissues (31.6％,28.8 ％,respectively,P ＜ 0.05).BRF2 was significantly associated with tumor differentiation,number of nodules,tumor encapsulation,TNM stage,and microvessel density.Prognosis of the high expression BRF2 group was poor.The recurrence and metastasis rates in the high expression group were significantly higher than the low expression group (P ＜ 0.05).The survival rate in the high expression group was significantly lower than the low expression group (P ＜ 0.05).Conclusions The expression of BRF2 increased in HCC tissues,and its expression was closely associated with pathological features and prognosis of patients,indicating that it can be used as a predictor in assessing prognosis of patients with HCC.

Clinical hematology and hematologic examination is a strong practical curriculum, and experimental class is very important in the aspects of teaching.In order to improve the quality of teaching and train compound laboratory talents with high quality,several effective reform mea-sures were carried out based on the years of teaching practice by the department:improving the methods of experimental teaching;using modern means of teaching;emphasizing on the analysis of experimental results,formulating the rigorous experimental evaluation system and starting the second class etc.

Objective To investigate the expression of Semaphorin3B (SEMA3B) in hepatocellular carcinoma (HCC) and reveal its clinical significance.Methods Immunohistochemistry and Western blotting were performed to examine the expression of SEMA3B protein in 56 hepatocellular carcinoma samples,matched perineoplastic tissues and 14 normal liver samples.Microvessel density (MVD) was determined by counting CD34-positive endothelial cells.Statistical methods were performed to analyse the relationships between the expression of SEMA3B and clinicopathologic factors.Results The rate of the SEMA3B-positive in HCC tissues was 42.9％ and significantly lower than that in normal liver tissues and non-cancerous adjacent liver tissues(78.6％ and 85.7％,respectively)(P＜0.05).MVD in SEMA3B-positive HCC tissus was lower than that in SEMA3B-negative HCC tissus (P＜0.05).The expression of SEMA3B was closely related to tumor nodular number,tumor size,tumor capsulation and CLIP score (P＜0.05 for all).The recurrence and metastasis rate in SEMA3B-positive group was lower than that in the negative group(P＜0.05),and the survival rate in positive group was higher than that in the negative(P＜0.05).Conclusion The expression of SE-MA3B was decreased in HCC tissues,and its expression was closely associated with tumor progression,angiogenesis and prognosis,indicating that it might seve as a predictor of prognosis and a possible novel target of antiangiogenic therapy for patients with HCC.

Objective To detect the immune effect of FbaAmAb2 against the surface protein of group A Straptococcus (GAS),and explore the pathogenesis and therapy of GAS infections.Methods By subclonal and bacterial ELISA,the positive hybridoma cells were screened that can produce better titers of FbaAmAb2 against GAS-surface FbaA protein,and were injected into the peritoneal cavities of BALB/c mice to produce ascites.The collected ascites were performed to dilute,as follows,original ascite,1:2,1:4,1:8,and 1:16 to test tube agglutination.Based on the results,we selected appropriate dilution to passively immunize mice,and then challenged the mice with GAS,evaluating FbaAmAb2 neutralizing ability with GAS in mice by the survival rate of the immunized mice.Whether FbaAmAb2 could inhibit the binding of factor H to GAS was confirmed by the invasive inhibition assay.Results The IgG titer of bacteria solution ELISA is 1:160 and the titer of tube agglutination is 1∶8.The protect rates of FbaAmAb2 on preventing mice with GAS infections are as follows:66.67％ in original ascite and 1:2 diluted groups,and 50％ in 1:4 diluted group.Mice in each experimental group were evoked significantly protective immune responses compared with the PBS control by SPSS analysis.FbaAmAb2 can competitively inhibit factor H binding to the surface proteins FbaA of GAS,which decreased the entry of GAS into the cytoplasm of human epithelial cells through the binding of factor H.Conclusion FbaAmAb2 is promising to be used in emergent prevention or the clinical therapy for GAS infection and it is promising starting points for pharmacologic targeting and further development of new therapeutic agents for GAS.

OBJECTIVETo construct a GFP/Puro double-labeled lentiviral expression vector for CK8 silencing and assess the effects of CK8 silencing on cell apoptosis.

METHODSThe siRNA sequences of CK8 were inserted into the lentiviral expression vector GV248 and transfected into 293T cells with the packaging plasmids PMD and SPA. The lentivirus was collected at 24 and 36 h post-transfection. Flow cytometry was used to detect the virus titer and the positive cells were selected with puromycin. The knockdown of CK8 was examined by Western blotting. The effect of CK8 down-regulation on cell apoptosis induced by cisplatin was detected with Annexin V/PI staining.

RESULTS AND CONCLUSIONWe successfully constructed CK8 interference lentiviral vector and obtained a stable cell line with CK8 knock-down that was sensitive to cisplatin-induced apoptosis.

Apoptosis ; Cell Line ; Down-Regulation ; Genetic Vectors ; Humans ; Keratin-8 ; genetics ; Lentivirus ; Plasmids ; RNA Interference ; RNA, Small Interfering ; Transfection

Objective:This paper is to propose a calibration model based on sine function which enables more choices to determine the functional relationship between the absorbance and the concentration of the tested substance.Methods:This paper uses Taylor series expansion and Levenberg-Marquardt to obtain the optimal parameters for the Sine model and then summarizes the characters of the Sine model. On the basis of these characters, this paper compares and evaluates the experimental data processed by the Sine model from four aspects: correctness, precision, linearity and correlation.Results:The generated sine function calibration model achieved deviations within ±3% of the national standard substance, precision ( CV%) less than 2%, and a linear correlation coefficient greater than 0.990 within the measurement range of 32-710 mg/L. The correlation coefficients between the sine model and other well-performing linear calibration models for 104 clinical samples were all greater than 0.990. Conclusions:The performance evaluation of the prealbumin assay kit using the sine function calibration model meets industry standards and shows good correlation with the results of clinical sample measurements. This indicates that the sine function calibration model can serve as a new calibration model for in vitro diagnostic research and clinical applications.

Eucommiae Folium(EF),a traditional Chinese medicine,has been used to treat secondary hypertension,including renal hypertension and salt-sensitive hypertension,as well as hypertension caused by thoracic aortic endothelial dysfunction,a high-fat diet,and oxidized low-density lipoprotein.The antihyperten-sive components of EF are divided into four categories:flavonoids,iridoids,lignans,and phenyl-propanoids,such as chlorogenic acid,geniposide acid and pinoresinol diglucoside.EF regulates the occurrence and development of hypertension by regulating biological processes,such as inhibiting inflammation,regulating the nitric oxide synthase pathway,reducing oxidative stress levels,regulating endothelial vasoactive factors,and lowering blood pressure.However,its molecular antihypertensive mechanisms are still unclear and require further investigation.In this review,by consulting the relevant literature on the antihypertensive effects of EF and using network pharmacology,we summarized the active ingredients and pharmacological mechanisms of EF in the treatment of hypertension to clarify how EF is associated with secondary hypertension,the related components,and underlying mechanisms.The results of the network pharmacology analysis indicated that EF treats hypertension through a multi-component,multi-target and multi-pathway mechanism.In particular,we discussed the role of EF tar-gets in the treatment of hypertension,including epithelial sodium channel,heat shock protein70,rho-associated protein kinase 1,catalase,and superoxide dismutase.The relevant signal transduction path-ways,the ras homolog family member A(RhoA)/Rho-associated protein kinase(ROCK)and nicotinamide adenine dinucleotide phosphate(NADPH)oxidase/eNOS/NO/Ca2+pathways,are also discussed.

OBJECTIVETo evaluate the effectiveness of small interfering RNA (siRNA) on inhibiting severe acute respiratory syndrome (SARS)-associated coronavirus replication, and to lay bases for the future clinical application of siRNA for the treatment of viral infectious diseases.

METHODSVero-E6 cells was transfected with siRNA before SARS virus infection, and the effectiveness of siRNA interference was evaluated by observing the cytopathic effect (CPE) on Vero-E6 cells.

RESULTSFive pairs of siRNA showed ability to reduce CPE dose dependently, and two of them had the best effect.

CONCLUSIONsiRNA may be effective in inhibiting SARS-associated coronavirus replication.

Animals ; Cercopithecus aethiops ; RNA, Small Interfering ; pharmacology ; SARS Virus ; drug effects ; Transfection ; Vero Cells ; Virus Replication ; drug effects

Objective : To explore the type and control measure of black dot⁃like contaminants in cell cultures. Methods: The growth state of bacteria was investigated under an inverted microscope ; Their morphological characteristics were analyzed by Gram and auramine O staining as well as electron microscopy; 16S rDNA gene sequencing was used to analyze bacterial species ; Drug sensitivity test was used to screen antibiotics against the bacteria;Cryopreserved SH⁃SY5Y cells were resuscitated by cell culture supernatant of RAW264 cells. Results: Inverted microscopic real⁃time observations showed that black dot⁃like substances had two growth states : static and moving.They were negative for Gram staining while positive for auramine O staining. Electron microscopy revealed that they were short rod⁃shaped bacteria with a polar flagellum during moving phase. 16S rDNA gene sequencing showed that these bacteria were phenylobacterium zucineum HLK1. Ceftriaxone , carboxycillin and imipenem were screened by drug sensitivity test to have inhibitory effects on the bacteria , but cell culture experiments showed that they could not remove the bacteria from SH⁃SY5Y cells. Contaminated cells could not be cryopreserved for a long time , but resuscitation with RAW264. 7 cell culture supernatant significantly improved the survival rate of cells. Conclusion The black dot⁃like contaminants in cell cultures are a special type of oligotrophic bacterium with strong viability that can invade the cells and cannot be cleared with antibiotic treatment. RAW264. 7 cell culture supernatant seems contain some substances against bacteria , and resuscitating frozen cells with RAW264. 7 cell culture supernatant may significantly improve the survival rate of cells.

【Objective】 To investigate the negative emotions among university students during home quarantine under the epidemic of coronavirus disease 2019 (COVID-19) so as to provide scientific basis for psychological counseling. 【Methods】 We conducted an online questionnaire survey to collect 405 undergraduates’ and postgraduates’ DASS-21 scale scores and responses to epidemic prevention knowledge, and analyzed the university students’ negative emotions during the epidemic and influencing factors. 【Results】 Of the 405 students surveyed, 178 individuals (44.0%) had depression, 171(42.2%) felt anxious, and 119(29.4%) felt stressed. Individuals in the DASS21+ group spent much time on entertainment on the cell phone daily than those in the DASS21- group (P<0.001), and the degree of impact by COVID-19 on daily life was significantly higher (P<0.001). Correlation analysis showed that entertainment time on the cell phone, the severity of COVID-19, and the impact of the epidemic were positively related to DASS-21 scores, with correlation coefficients of 0.231, 0.143 and 0.259, respectively (all P<0.01). Binary logistic regression analysis found that mobile entertainment time for over 5 hours per day (OR=3.370, 95% CI: 1.369-8.294, P=0.008) was a risk factor for negative emotions during quarantine at home. 【Conclusion】 During quarantine at home in the epidemic of COVID-19, nearly half of university students are prone to breed negative emotions, like depression, anxiety or stress, which may be related to long-term mobile phone entertainment.