Herpes simplex virus type 1 (HSV-1) is the causative agent of cold sores and other more serious diseases. HSV-1 infected-cell protein 27 (ICP27) is an immediate-early regulatory phosphoprotein homologous to gene products identified in all classes of herpesviruses so far. To raise the antiserum to ICP27 for further characterization of its biological function, the ICP27 gene was cloned into the pET-28a (+) vector, then ICP27 protein was expressed in E. Coli and purified by nickel-nitrilotriacetic acid (Ni2+-NTA) affinity resin column,finally the purified protein was used to raise antiserum. Western blot analysis demonstrated that the antiserum recognized the recombinant protein, and the antiserum was able to probe the ICP27 in HSV-1 infected cells with high specificity by immunofluorescence assay (IFA). Therefore, the specific antiserum will provide a valuable tool for further studies investigating ICP27's biological function during HSV-1 infection.

Objective To study the action of CNOT 7 (CCR4-NOT transcription complex subunit 7 human)gene and its mechanisms in the process of Vγ 9Vδ2T cell immunologic tolerance of HepG2 cells (Hepatoblastoma G2 Cell Line).Methods The shCNOT 7 (Recombinant plasmid of CNOT 7) and control vector shRNA were transfected into HepG2 cells.Vγ9Vδ2T cytokine stimulated each group before and after cell transfection,Cell apoptosis was detected by flow cytometry (FCM),CNOT 7 protein and STAT1,STAT3 expression level was detected by Western blot.CNOT 7,STAT1 and STAT3 protein expression levels of HepG2 liver cancer cell lines and L02 normal liver cell line was assayed by Western blot.Results When stimulated by Vγ9Vδ2T cytokine,the apoptosis rate of gene-knockdown group significantly improved from (7.55％ ±2.63％) to (20.59％ ±3.12％).Compared with L02 cells,the CNOT7 protein expression of HepG2 cells increased (F =28.76,P ＜ 0.01),STAT3 protein expression increased (F =110.29,P ＜ 0.01),while STAT1 protein expression was down-regulated (F =35.67,P ＜ 0.01).CNOT 7 knockout could induce HepG2 cells STAT1 expression (t =6.69,P ＜ 0.05).Conclusions CNOT 7 gene could induce HepG2 cells Vγ 9Vδ2T cellular immune tolerance.CNOT 7 knockout could reverse the Vγ 9Vδ2T cell immunologic tolerance of HCC.

To obtain recombinant human β2-microglobulin (rhβ2M) with properties of good solubility and high purity from E. coli, prokaryotic expression conditions were optimized and protein purification was performed in this study. After testing the effect of different IPTG concentrations, temperatures and induction times on the production of rhβ2M, the optimum expression conditions were determined, i. e. joining IPTG to final concentration being 0.8 mmol/L and inducing time 6 h and at temperature of 25 degrees C. Under the optimum induction conditions, the ratio of soluble rhβ2M to soluble bacterial protein was 63.7%. After purified by Ni Sepharose 6 Fast Flow, the purity of rhβ2M achieved a greater value of 95%. Western blot analysis revealed that rhβ2M possessed the antigen property that specifically interacted with anti-β2M antibody.
Blotting, Western ; Escherichia coli ; metabolism ; Humans ; Recombinant Proteins ; biosynthesis ; Solubility ; beta 2-Microglobulin ; biosynthesis

Objective To survey and control an extensively drug-resistant Acinetobacter baumannii (A.bauman-nii)(XDRAB)lower respiratory tract infection (LRTI)outbreak in intensive care unit (ICU)of a hospital. Methods From October 27,2013 to December 27,2013,5 cases of XDRAB LRTI occurred in ICU,epidemiological investigation was conducted,sources and transmission routes of infection were searched,intervention measures were performed.Results XDRAB were isolated from patients’sputum and environmental object surface,drug-resistant spectrums of isolated XDRAB were almost the same,suggesting the outbreak was due to the contamination of envi-ronment by the same pathogen.54 specimens were taken before disinfection,26 were positive,XDRAB were isola-ted from door handles,sheets and telephones.Investigation concluded that the outbreak of XDRAB infection was due to contamination of indoor environment and equipment,incomplete disinfection,and inadequate hand hygiene compliance of health care workers.After the implementation of a series of control measures,XDRAB was not found,and there was no new XDRAB infection cases.Conclusion Environment contamination is the main cause of this XDRAB healthcare-associated infection(HAI)outbreak,strict implementation of isolation,prevention and con-trol measures of MDROs can effectively control HAI outbreak caused by A.baumannii.

Objective To observe the changes of pulmonary function parameters in healthy middle-aged and elderly people in Shanghai.Methods A total of 298 subjects aged 45 years and over with normal pulmonary function and no smoking history were selected among subjects who underwent routine physical examination from Jun 2009 to Sep 2011.The subjects were divided into three groups:45-59 years,60-69 years,and ≥70 years.The tested pulmonary function included 13 parameters:vital capacity (VC),residual volume (RV),functional residual capacity (FRC),total lung capacity (TLC),RV/TLC,forced vital capacity (FVC),forced expiratory volume in one second (FEV1),FEV1/FVC,peak expiratory flow (PEF),forced expiratory flow at 25 ％ of FVC exhaled (FEF25),forced expiratory flow at 50％ of FVC exhaled (FEF50),diffusion capacity of the lung for carbon monoxide (DLCO),and specific diffusion capacity of CO (KCO).Results Compared with the young adults,7 pulmonary function parameters including VC,FVC,FEV1,PEF,TLC,DLCO and KCO were declined significantly in healthy middle aged and elderly people Compared with the young adults,pulmonary function parameters including RV and RV/TLC were increased [male (1.9 ±0.4)Lvs.(1.6±0.3)L,(34.7±4.9)％ vs.(26.4±4.3)％,P＜0.01;female (1.5±0.3)L vs.(1.3±0.3)L,(36.1±5.3)％ vs.(30.3±5.5)％,P＜0.01].Although RVincreased with aging in middle-aged and elderly people,but there was no statistical significance (P＞0.05).FRC didn't change obviously.FEV1/FVC was lower in old people than in young people[male (80.7±4.3)％ vs.(84.2±5.6)％,P ＜0.01;female (82.5± 5.5) ％ vs.(86.4±4.8)％,P＜0.01],but the difference between 3 groups of middle-aged and the elderly was not significant.The FEV1/FVC of men and women aged 70 and more were (80.2±4.5)％ and (81.8±5.6)％,respectively.Conclusions Both ventilation function and diffusion function of healthy elderly people are decreased with ageing,RV is increased and FRC is steady,and the declining tendency of FEV1/FVC obviously slows down with aging in middle aged and elderly people.

Pancreatic cancer is one of the most malignant tumors with a high mortality rate attributed to its widespread metastasis.A number of cellular signal transduction pathways involved in multiple genes play an important role in regulating this complex metastatic cascade of pancreatic cancer.NF-kappa B is one of the crucial signaling pathways.Studies has indicated that NF-kappa B could modulate a series of biological events relevant to tumor progress by controlling multiple targeted genes expression,such as cell proliferation,anti-apoptosis,angiogenesis,epithelial-mesenchymal transition,inflammation,stress response,etc.Furthermore,it can also up-regulate Hedgehog and MMPs signaling pathways.To help us better understand the potential mechanism and identify more sensitive tumor markers and selective targets,this review will underline the significant roles of NF-kappa B signaling pathway in regulatory network of pancreatic cancer metastasis.

Objective To evaluate the effect of the combination of dexmedetomidine and sufentanil on monitoring anesthesia care during burr-hole surgery for patients with chronic subdural hematoma. Methods 96 pa-tients underwent burr-hole surgery for chronic subdural hematoma with MAC were randomly divided into two groups:Group D and Group DS (n=48 in each group). Local anesthetic block was started at least 10 min after DEX and sufentanil infusion. Ramsay sedation scale of the two groups was maintained to 3. Anesthesia onset time, hemo-dynamics, the amount of rescue midazolam or fentanyl, the time to first dose of rescue midazolam or fentanyl, the to-tal number of intraoperative patient movements, postoperative recovery time, patient and surgeon satisfaction scores, and the adverse events were recorded. Results Compared with group D, anesthesia onset time was significantly less in group DS (13.68 ± 3.13 vs. 11.82 ± 2.43 min, P=0.002). More patients in group D required rescue midazol-am to achieve RSS=3 compared with group DS (31.25%15/48 vs. 12.50%6/48, P=0.023). Compared with group D, significantly fewer patients in group DS required rescue fentanyl to relieve pain (10.42%5/48 vs. 27.08%13/48, P = 0.036). Additionally, the total dose of rescue fentanyl in group DS was significantly higher (89.48 ± 23.27 vs. 125.28 ± 33.52μg, P=0.000), and the time to first dose of rescue fentanyl was longer than group D(18.34 ± 4.45 vs. 14.34 ± 3.63 min, P=0.000). The total number of patient movements during the burr-hole surgery was higher in group D than group DS (35.42%17/48 vs. 16.67%8/48, P=0.036). The time to recovery for discharge from the PA-CU (time to an Aldrete score ≥ 9) was significantly shorter in group DS compared with group D (17.54 ± 5.92 vs. 12.57 ± 5.28 min, P=0.000). Results from the patient and surgeon satisfaction scores showed significant differenc-es favoring group DS (P<0.05). More patients in group D showed higher levels of the overall incidence of bradycar-dia (37.50% 18/48 vs. 18.75% 9/48, P = 0.041) and hypotension(37.50%18/48 vs. 14.58%79/48, P=0.011)com-pared with group DS. Conclusions Compared with DEX alone, DEX-sufentanil associated with fewer number of in-traoperative patient movements, less amount of rescue scheme, could be safely and efficiently used for MAC during burr-hole surgery for patients with chronic subdural hematoma.

Objective To establish a stable animal model of transplanted human colorectal cancer.Methods BALB/c nude mice were randomly divided into orthotopic colorectal model group and subcutaneous inoculation model group by random number table,and separately inoculated with 0.1 ml human colon cancer cell HCT116 under the density of 2×107/ml into the orthotopic colorectal (with the self-made inoculator) and right forelimb pit subcutaneously.The mice were observed for 60 days to compare the tumor formation and tumor growth in the two groups.Results The tumor formation rate of all 18 animals was 100 ％ (18/18) in the orthotopic colorectal group.The average tumor weight was (2.78±1.86) g and the average survival time was (45.00±11.99) d.The tumor formation rate was 27.78 ％ (5/18) in the subcutaneous inoculation group.The average tumor weight was (1.74±0.82) g,and the average survival time was (60.00±0.00) d.Conclusion 0.1 ml (2×107/ml) human colon cancer cell suspension HCT116 inoculated into BALB/c nude mice orthotopic colorectal with self-made inoculator could establish human colorectal cancer animal model successfully.