Objective To construct and identify the specific interference vector for second mitochondria -derived activator of caspase (SMAC)gene in mice and perform lentiviral packaging.Methods According to the SMAC gene sequences in mice,three small interfering RNAs (siRNAs)(siRNA1,siRNA2,and siRNA3)and one negative control sequence (siRNAn)were designed and synthesized,and mouse hepatocytes were transfected with the above siRNAs using Lipofectamine 2000.The inhibitory effects of these siRNAs on SMAC mR-NA expression were evaluated by real-time PCR,and the optimal siRNA was screened out accordingly.Oligo DNA was synthesized based on the optimal siRNA and then connected to GV1 15 vector to obtain recombinant interference plasmid.The candidate clones were identified by PCR and sequenced.The recombinant interference plasmid,as well as pHelper 1.0 and pHelper 2.0,was used to transfect 293T cells for lentivirus packaging.Then,cell supernatants were collected and concentrated,and the lentiviral titer was determined by gradient dilution. Comparison between groups was made by analysis of variance,and multiple comparisons were made by SNK-q test.Results The three siRNAs had different inhibitory effects on SMAC mRNA expression in hepatocytes;siRNA1 showed the strongest inhibitory effect,with an inhibition efficiency of 70.3%.Based on siRNA1 ,the oligo DNA was successfully synthesized and correctly connected to the lentiviral vec-tor,as proved by sequencing.The lentiviral titer was 6 ×108 TU/ml,as determined by gradient dilution.Conclusion The recombinant lentivirus that inhibits SMAC expression in mice has been constructed successfully,laying the foundation for further studies on the role of SMAC gene in liver failure.

Oxidative damage theory is currently one of the predominant theories on the mechanisms of aging. Previous research has shown that antioxidants can extend the lifespan in the model organism by scavenging free radicals,inducing the expression of stress related genes and hormesis. However, recent studies have suggested that these pharmaceuticals may cause serious side effects,such as promoting oxidation,increasing the risk of cancer,and destroying the metabolic balance. The low absorption and targeting property also limit the efficiency of most antioxidants. As a result ,the correlation between antioxidants and lifespan extension remains to be demonstrated. We reviewed the research progress in the field of lifespan extension by antioxidants in recent years and provided references for future research in related areas.

Objective To discuss the regulatory effect of 5-aza-2 ,-deoxycytidine on P16 and MGMT in cervical cancer cells. Methods After four kinds of cervical cancer cells (HeLa, SiHa, C33A and CaSki) were treated with 5-Aza-dC , MSP was used to detect the methylation variation of P16 and MGMT , and fluorogenic quantitative PCR and Western blot were employed for determination of P16 and MGMT expression. MTT and Annexin V-FITC/PI double staining were adopted for detection of cell proliferation and apoptosis. Results Both P16 and MGMT exhibited methylation in four kinds of cervical cancer cells , and after treatment with 5-Aza-dC ,their methylation levels were reversed. 5-Aza-dC was able to inhibit p16 and MGMT expression in the cervical cancer cells, and can also suppress cell proliferation and promote apoptosis. Conclusions Although methylation of P16 and MGMT are present in cervical cancer cells, their expression level was still high. Therefore, regulation of P16 and MGMT expression may be affected by other factors. 5-Aza-dC can suppress the growth of cervical cancer cells. Although 5-Aza-dC reverse the methylation levels of P16 and MGMT, it inhibits their gene expression. More experiments are needed to verify the hidden reasons and mechanisms.

Cell-penetrating peptides (CPPs) offer a non-selective and receptor-independent mode to promote cellular uptake. Although the non-specificity of CPP-mediated internalization allows this approach applicable to a wide range of tumor types potentially, their universality is a significant obstacle to their clinical utility for targeted delivery of cancer therapeutics and imaging agents. Accordingly, many reports have focused on selective switching of systemically delivered inert CPPs into their active form in lesions (tumor). In this review, our attention is mainly confined to such an enzyme-sensitive domain incorporated delivery system with activatable CPPs (ACPPs), which have displayed the exciting strength in balancing the CPPs' pros and cons, and potential in the treatment and diagnosis of some diseases.

Objective To research the molecular biology characteristics and transient expression in BHK-21 cells of E geue segment from Japanese encephalitis virus(JEV) and construct an eukaryotic expression vector pEGFP-C1-JEV.Methods E gene segment of JEV was amplified by RT-PCR,construct the recombinant vector pEGFP-C1-JEV,which could express EGFP label proteins.Transfect pEGFP-C1-JEV vector into BHK-21 via LipofectAMINETM 2000,to observe expressing of EGFP label protein and transcription of aim gene,and to check up localization and antigenicity of expressed E protein by Immunohistochemistry and Western blot.Results It showed that the recombinant plasmid pEGFP-C1-JEV was successfully constructed and transfected to BHK-21 cells,the normal expression of green fluorescent protein expression rate was higher.RT-PCR showed that gene transcription in BHK-21 and normal expression,expression protein was mainly distributed in the cytoplasm of BHK-21 cells and the envelope in,and can with guinea pig anti-JEV antibody binding.Conclusion pEGFP-C1-JEV vector in BHK-21 cells was normal expression and there were no effect on cell growth and morphology.Meanwhile,on eukaryotic antigens was good antigenicity.This research as a base foundation for E protein gene of JEV eukaryotic expression and function in vitro and applied research.

Performance appraisal of public hospitals have given a guidance for the development of public hospitals at all levels.A Class A tertiary hospital reviewed the problems in the development of the hospital at the present stage and focused on the following four aspects:①insufficient fine management;②No clear orientation of discipline development;③The bottleneck of the improvement of medical operation efficiency;④New challenges in the reform of payment mode.The tertiary hospital launched a fine management practice in May 2022,in order to solve the problems by taking the Department of Surgery as a pilot area,laying the foundation for fine management through information system construction,improving the efficiency of medical operation through management process optimization,improving the overall competitiveness of disciplines through the construction of sub-specialty and Discipline Alliance and adjusting the performance appraisal index system to play the role of performance incentives.The measures effectively improve the overall capacity and efficiency of hospital medical services and help the hospital to achieve high-quality development.

Autologous ozonized blood transfusion(AOBT) is a therapy of re-transfusion of 100-200 mL of autologous blood after shaking and agitation with appropriate amount of oxygen-ozone in vitro. The oxidation of blood through the strong oxidation of ozone can enhance the non-specific immune response of the body, regulate the internal environment and promote health. This therapy has been increasingly applied in clinical practice, while no unified standard for the operation process in terms of ozone concentration, treatment frequency and treatment course had been established. This operation process of AOBT is primarily explored in order to standardize the operation process and ensure its safety and efficacy.