Animals ; Apoptosis ; physiology ; Carcinoma, Squamous Cell ; drug therapy ; physiopathology ; prevention & control ; Cyclooxygenase 2 ; Cyclooxygenase 2 Inhibitors ; Cyclooxygenase Inhibitors ; therapeutic use ; Head and Neck Neoplasms ; drug therapy ; physiopathology ; prevention & control ; Humans ; Membrane Proteins ; Neoplasm Invasiveness ; Neoplasm Metastasis ; drug therapy ; Neovascularization, Pathologic ; drug therapy ; metabolism ; Prostaglandin-Endoperoxide Synthases ; metabolism

The three immediate-early proteins of HSV-1, ICPO, ICP22, and ICP27, have specific and pivotal functions in transcriptional activation and inhibition, multiple regulatory and control processes of viral genes. In this paper, the expression and localization of these three proteins were studied in neuroblastoma cells using biochemical assays, and their possible and potential interactive functions are discussed. The data show that the three proteins are localized in different structures, specifically in the PML-NB-associated structure, which is a specific nuclear structure composed of many protein molecules and bound tightly to the nuclear matrix in neuroblastoma cells. The results suggest that the activating and suppressive functions of ICPs are mostly dependent on their transcriptional and regulatory roles, including the PML-NB-associated structure.

Different degrees of HCV re-infection exist in patients with hepatitis C after liver transplantation. Its pathogenesis is different according to different phases of the disease. Factors affecting its recurrence include HCV gene type, viral load, HLA matching between donor and recipient, time of recurrence, donor's age and so on, in which the application of immunosuppressants is the most important influencing factor. The virological response can be used to evaluate the effects of treatment. Now, it is widely accepted that the best choice and therapeutic plan is Pegy interferon alfa-2a/2b combined with Ribavirin.

Objective Summarizing single clinical experience with extracorpomreal membrane oxygenation(ECMO) as a supplement to extracorporeal cardiopulmonary resuscitation(ECPR) in adult patients with cardiac arrest to explore new ideas.Methods We retrospectively analyzed the characteristics of 17 patients who underwent ECMO as part of ECPR from July 2005 to September 2014 at Fuwai Hospital,and analyzed the differences between the survival group(n =6) and the in-hospital death group.Results The mean CPR time was(44.53 ± 21.39) min.The support duration of ECMO was(106.38-± 70.43) h.12 patients of all were successfully weaned from ECMO,and 6 patients survived to hospital discharge.There were significant differences between the two groups in terms of the last serum creatinine and blood lactate acid level before ECMO,and the time to lactate normalization.11 patients died,7 patients developed bleeding,and 8 cases developed infection.Conclusion Single-center data showed that applying ECMO as a means of ECPR improved the survival rate in cardiac arrest patients.Additionally,creatinine and lactic acid were good indicators for assessing prognosis.Refractory circulatory dysfunction and neurologic complications have an adverse impact on the survival of cardiac arrest patients.

Herpes simplex virusⅠ(HSVⅠ) regulating the pathway of transcription and translation modify in host cell is a very systematic and complicate system. A clear understanding of the concrete mechanisms of infection will greatly help to comprehend the virus replication and the interaction with the host cell. By the analysis of 2-DE, the heterogeneous nuclear ribonucleoprotein H2 in human fetal liver cell represent distinction after the HSVⅠinfection.Utilization of Northern blot and Western blot technologies verified the expression of hnRNP H2 in different stage of virus infection is varied.

HIV-1 envelope glycoprotein gp41,which is a hopeful target for HIV-1 fusion inhibitors,plays a critical role in the fusion of viral and cellular membranes.In order to build up the screening assay of HIV-1 fusion inhibitors targeting gp41,HIV-1 gp41 5-helix and 6-helix were expressed in prokaryotic cells.Gp41 5-helix and 6-helix recombined plasmids were constructed by using PCR,enzyme digestion and ligation taking the clade B HIV-1 genome as a template.The plasmid was transferred into E.coli BL21(DE3)and then induced by IPTG.The expressed protein was purified by affinity chromatography after denaturation and renaturation.The SDS-PAGE analysis was used during expression and purification.Native-PAGE was used to identify the interaction between gp41 5-helix and T-20.The result will be helpful to build up the screening assay of HIV-1 fusion inhibitors targeting gp41.

Highly Active Anti-Retroviral Therapy (HAART) has effectively inhibited the prevalence of HIV-1 and reduced the death rate caused by AIDS. In recent years,the emergence of resistance-conferring RT gene mutations in HIV-1 strains has become the major reason for HAART failure. The detection of drug resistance is important for the HAART regimen choice and novel drug development. A novel assay for the detection of HIV-1 RT drug resistance mutations was developed. HIV-1 drug resistance and wild strains in B subtypes were investigated using Two-Step Mutagenically-Separated PCR (MS-PCR),and point mutations including M41L,K70R,K103N,Y181C,T215F were detected. A longer mutant type primer was designed,using microplates hybridization and ELISA technique to detect several point mutations within a mixed mutant-wild type population. The results indicate that the Two-Step MS-PCR is as sensitive and specific as that in the traditional MS-PCR and MS-PCR combined with ELISA can give a good P/N quotient with better sensitivity,low cost,relatively less time consumption and high-throughput screening. It will be used in clinic usage for the detection of HIV-1 drug resistance mutations as well as other point mutations.

OBJECTIVETo explore the mechanism of polypeptide extract from scorpion venom (PESV) on inhibiting angiogenesis.

METHODSThe H22 hepatoma tumor model was established by subcutaneously implanting H22 hepatoma cells into mice. The tumor-bearing mice were randomly divided into 4 groups, i.e., the control group, the high dose PESV group, the low dose PESV group, and the 5-fluorouracil (5-Fu) group, 10 mice in each group. The intervention was lasted for 14 days. The growth curve of the tumor volume was drawn and the inhibition rate calculated. Pathological changes of the tumors were observed by HE staining. The microvessel density (MVD) was detected using SP method. The protein expression levels of phosphatidylinositol 3-kinase (P13K), phosphoprotein kinase B (P-Akt), hypoxia-inducible factor-1 alpha (HIF-1 )alpha, and vascular endothelial growth factor-A (VEGF-A) were detected by immunohistochemical assay and Western blot.

RESULTSThe tumor inhibitory rate was 64.8%, 43.7%, and 32.4% in the 5-Fu group, the high dose PESV group, and the low dose PESV group. Compared with the control group, the protein expression of PI3K, P-Akt, HIF-1alpha, and VEGF-A were obviously inhibited by PESV and 5-Fu (P <0. 05,P <0. 01). The MVD also decreased in the high and low dose PESV groups (P < 0.05).

CONCLUSIONSPESV could inhibit the angiogenesis of H22 hepatoma. The mechanisms might be associated with suppressing the expression of PI3K, P-Akt, HIF-1 alpha, and VEGF-A.

Angiogenesis Inhibitors ; pharmacology ; Animals ; Cell Line, Tumor ; Fluorouracil ; pharmacology ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Liver Neoplasms ; Male ; Mice ; Peptides ; pharmacology ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Scorpion Venoms ; pharmacology ; Vascular Endothelial Growth Factor A

The pol gene of HIV-1 encodes mainly three enzymes: reverse transcriptase (RT), protease (PR) and integrase (IN). Currently, FDA approved drugs targeting RT and PR are available and administered in various combinations, while no anti-IN drug was approved. HIV-1 integrase is an essential enzyme for the viral replication and an interesting target for the design of new pharmaceuticals for multi-drug therapy of AIDS. The 288 amino acids of IN (32kDa) recognizes specific sequences in the long terminal repeats (LTRs) of the retroviral DNA. The IN protein catalyzes the 3′-processing step and the 5′-strand transfer step reaction in vivo, which was called integration and this reaction could be analysed by ELISA Assay in vitro. It has been reported that F185K and C280S mutations of HIV-1 integrase would improve the enzyme solubility, and the catalytical activity of the enzyme was the same as that of the wild-type enzyme in vitro. In order to build the platform of screening inhibitor against integrase of HIV-1 virus, the IN enzyme was expressed and the function of integrase protein was assayed. The cDNA of clade B HIV-1 genome was used as a template, overlapped PCR was used to construct site mutagenesis of F185K/C280S and NdeI/Xho I restriction sites were brought in. The PCR product was cloned into the prokaryotic vector pET-28a(+) to form a recombined plasmid, transferred into the host cell E.coli(BL21 DE3). The recombined clones were identified by PCR and Nde I/Xho I digestion .The positive plasmid was sequenced, and the successfully recombined plasmid in the host cell was induced by IPTG. The expressed IN protein was puriied sy the Co+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombined IN protein. The recombinant protein was soluble, and expressed highly and stably in E.coli. The molecular weight of the expression product was identical to the expectation. The IN protein was proved to be functional in 3′ processing and 5′strand transfer by ELISA. It will be helpful to build the platform of screening inhibitors against HIV-1 integrase.

Objective:To explore the application effect of 5A nursing model based on WeChat platform in patients with chronic obstructive pulmonary disease (COPD).Methods:A total of 82 patients with COPD were enrolled in our hospital from February 2018 to March 2019. According to the order of establishment, the patients were divided into study group and control group, with 41 cases in each group. The control group received routine care, and the study group adopted a 5A care model based on the WeChat platform on the basis of the control group. Pulmonary function before and after intervention in the two groups [1 second forced expiratory volume (FEV1), forced vital capacity (FVC), FEV1/FVC)], mood disorder [Self-rating Anxiety Scale (SAS), Depression Self-rating Depression Scale (SDS)], self-efficacy [General Self-Efficient Energy Meter (GSES)] supplements the Chinese name of this scale), quality of life [St. George Respiratory Questionnaire (SGRQ)] supplements the Chinese name of this scale) and nursing job satisfaction.Results:The FEV1, FVC and FEV1/FVC of the study group were (0.98±0.07)L, (1.59±0.22)L, (61.64±1.88)%, and the control group was (0.99±0.08)L, (1.61±0.20). L, (61.49±2.05)%; after 3 months of intervention, the study group was (1.45±0.18)L, (2.01±0.24)L, (72.14±1.49)%, and the control group was (1.21±0.12)L, (1.83±0.21) L, (66.12±1.75)%, there was no significant difference between the two groups before the intervention of FEV1, FVC, FEV1/FVC ( t value was 0.602, 0.431, 0.345, P>0.05), intervention 3 The difference between the two groups was statistically significant ( t value was 7.104, 3.614, 16.771, P<0.05). The SAS and SDS scores of the study group were 59.17±3.25 and 61.02±4.06; and 58.96±3.72, 60.75±3.84 in the control group; after 3 months of intervention, the study group scored 42.05±3.10, 44.26±3.25, and the control group scored 48.59±3.55, 49.97±3.41. There was no significant difference in the scores of SAS and SDS between the two groups before intervention ( t value was 0.272, 0.309, P>0.05). After 3 months of intervention, the difference between the two groups was statistically significant ( t value was 8.885, 7.762, P<0.01); the GSES score of the study group was 22.19±3.11 before the intervention, and the control group scored 22.58±2.97. After 3 months of intervention, the study group scored 33.65±2.14. The scores of the group were 28.71±2.03. There was no significant difference in the GSES scores between the two groups before intervention ( t value was 0.581, P>0.05). After 3 months of intervention, the difference between the two groups was statistically significant ( t value was 10.724, P<0.01). Conclusion:The 5A nursing model based on WeChat platform can significantly improve lung function, reduce mood disorder, enhance self-efficacy, improve quality of life, and patients showed high satisfaction with nursing work.