Hepatitis B virus (HBV) is a major cause of chronic liver disease, and frequently results in hepatitis, cirrhosis, and ultimately hepatocellular carcinoma. HBV polymerase (Pol) is an essential viral protein that is important for HBV replication and might be involved in the development of hepatocellular carcinoma. Protein-protein interactions appears to be crucial for its role. The aim of this study was to screen and identify the proteins that interact with Pol using a co-immunoprecipitation-based LC-MS/MS identification technique. The HBV Pol gene was amplified by polymerase chain reaction (PCR) and cloned into pCDNA3.1(+). The recombinant plasmid pCDNA3. 1(+)-Pol-flag was transfected into HeLa cells. Liquid chromatography and tandem mass spectrometry (LC-MS/MS) identified 45 proteins that co-immunoprecipitated with flag-tagged HBV Pol. Eleven of these have previously been reported as proteins that interact with HBV Pol. A proof-of-concept-based Ingenuity Pathway Analysis (IPA, www.ingenuity.com) was used to characterize the functions and pathways of these 45 identified proteins and HBV Pol. Among these proteins, four proteins may play a role in three major molecular cellular networks, and are therefore worthy of further investigation.
Cell Line, Tumor ; Chromatography, Liquid ; methods ; Gene Products, pol ; chemistry ; genetics ; metabolism ; Hepatitis B ; genetics ; metabolism ; virology ; Hepatitis B virus ; chemistry ; enzymology ; genetics ; Humans ; Immunoprecipitation ; methods ; Protein Interaction Maps ; Software ; Tandem Mass Spectrometry ; methods

To know the effect of subconjunctival injection of liposome encapsulated cytarabine on proliferation of conjunctival fibroblasts, the conjunctiva was isolated at 180 degrees from the injection site 3 days after subconjunctival injection of the normal saline (control), cytarabine, liposome encapsulated cytarabine, and 1 day after injection of cytarabine, and then those were inoculated in the culture media of fibroblasts. In the case of 3 days after injection of cytarabine, there was 49% and 42% inhibition of proliferation of conjunctival fibroblasts compared with the control respectively. Therefore, the authors concluded that the liposome encapsulated cytarabine is effective on inhibition of proliferation of conjunctival fibroblasts and reduces the frequencies of subconjunctival injection compared with the cytarabine itself.
Conjunctiva ; Culture Media ; Cytarabine* ; Fibroblasts* ; Liposomes*

AIM: To evaluate the ability of spectral domain optic coherence tomography ( SD-OCT ) parameters to detect progressive structural damage in primary open angle glaucoma ( POAG) by contrasting with visual field. ·METHODS: Retrospectively we evaluated 48 subjects (48 eyes ) of POAG, followed up 14 to 62mo. The parameters of SD-OCT and visual field were obtained. The correlation between the change of visual field measurements and OCT measurements were analyzed. Visual field progression was defined as reproducible drop of at least 2dB of mean deviation (MD) from the baseline visit. ROC curve was made to evaluated the ability of OCT parameters in detect progression of POAG. ·RESULTS: The 25 eyes were classified in progression group and 23 eyes were in non-progression group. No significant correlation was seen between OCT parameters and visual field in non-progression group. In progression group, OCT parameters which were significantly correlated with MD reduce were rim volume ( r=-0. 5997, P=0. 0007), C/D vertical ratio (r=-0. 6309, P=0. 0003), RNFL(r= 0. 4201, P= 0. 0260), and GCC(r= 0. 7080,P<0. 01). ROC curves showed the GCC reflected the reduce of MD accurately (P=0. 013). ·CONCLUSION: Part parameters of SD-OCT can reflect the progression of POAG accurately and provided a new method to detect the damage of visual function of POAG.

OBJECTIVETo evaluate the outcome of Titanium elastic intramedullary nailing (TEN) for the treatment of displaced midclavicular fractures.

METHODSBetween February 2010 and February 2013, 62 patients with displaced midclavicular fractures were treated by TEN, including 27 males and 35 females with an average age of 37.6 years old ranging from 15 to 67 years. The course of disease was from 1 to 9 days (means 2.7 days). Thirty-nine cases were treated by closed reduction and 23 cases by assistant small incision. Based on OTA (the Orthopaedic Trauma Association classification) classification, 31 cases of simple fractures involved 5 cases of 06-A1,15 cases of 06-A2,11 cases of 06-A3; 31 cases of wedge fractures involved 4 cases of 06-B1,12 cases of 06-B2,15 cases of 06-B3. Postoperative pain relief were evaluated by VAS score,and operation time,fracture healing time were recorded. After 6 weeks and after removing internal fixation shoulder joint function was evaluated by Constant score, and shoulder joint function were assessed by Herscovici score after 6 weeks.

RESULTSExcept 2 cases were lost to follow-up, 2 cases did not remove internal fixation, 3 cases' fractures were nonunion, the remaining 55 patients received follow-up for an average time of 11.4 months. The average preoperative VAS score was 5.20±0.71, and it turned to be 1.550.59 at 3 days after operation. The average operative duration was 40 min (15 to 65 min). The average bone healing time was (2.71±0.54) months (2 to 5 months). No difference about bone healing time was found between simple fractures and wedge fractures,and between the patients treated by closed reduction and assistant small incision. According to Herscovici standard, the shoulder function was excellent in 49 cases, good in 4, fair in 1, and poor in 1. Simple fractures achieved better Constant score than wedge fractures at 6 weeks postoperatively, whereas no difference was found at 4 weeks after fixation removal.

CONCLUSIONTitanium elastic nails (TEN) for treatment of displaced midclavicular fractures has advantages of good clinical effect,rapid pain relief,fast recovery of shoulder joint function. For OTA classification type B patients with comminuted fracture can replace steel to achieve very good effect, but to grasp the appropriate.

Adolescent ; Adult ; Clavicle ; injuries ; surgery ; Female ; Fracture Fixation, Intramedullary ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged

Objective To monitor the expression patterns of bcr-abl in chronic myeloid leukemia (CML) patients during treatment with imatinib mesylate and evaluate the detection of MRD by RQ-PCR method. Methods The ABI Prism 7500 Sequence Detection System using Taqman fluorogenic probes was used to quantify target gene. bcr-abl mRNA was detected by RQ-PCR in 106 CML patients. The normalized quotient (NQ) of bcr-abl mRNA was calculated as followings: NQ=bcr-abl mRNA copy numbers/abl mRNA copy numbers. Results The NQ of BCR-ABL mRNA was well correlated with the progression of disease and the number of Ph+ cell (r =0.9824 and 0.9346, respectively). The NQ was decreased rapidly in 62 patients and kept in low level for a long time, and only 2 of them were relapsed. For 8 patients, after treatment the NQwere decreased initially and increased sharply, 7 of them were relapsed after 5-9 months. After treatment the NQ of 31 patients were still>0.1, 11 patients were relapsed after a short remission and 7 were ineffective or progression. Out of 5 patients whose NQ were fluctuated and had little regularity, but all of them had a continuing remission. Conclusion RQ-PCR is a more sensitive technique in the detection of bcr-abl fusion gene.It is an important method to monitor the tumor cell during the treatment with imatinib mesylate in CML patients.

Objective:To investigate the role of extracellular signal-regulated kinase 1/2(ERK1/2)signal pathway in the pathogenesis of stress-induced gastric ulcer.Methods:Animal model of stress-induced gastric ulcer was established in rats with water-immersion restraint(WIR)stress.The mucosal activation of ERK1/2 was observed before and 5,15 and 30 min,and 1, 2 and 3.5 h after WIR stress.Some animals were also treated with an intravenous injection of PD98059(1 mg/kg),a specific ERK1/2 inhibitor,1 h prior to WIR stress.Expression of total ERK1/2 and caspase-3 were detected by Western blot analysis; ERK1/2 activity was measured by kinase activity assay using myelin basic protein as a specific substrate.DNA-binding activities of the transcription factors activator protein-1(AP-1)and nuclear factor-?B(NF-?B)were determined by electrophoretic mobility shift assays(EMSA).Mucosal TNF-?and IL-1?mRNA expression was analyzed by Northern blot analysis.The degrees of the gastric mucosal lesions were expressed as ulcer index(UI)and pathological evaluation.Apoptosis in the gastric mucosa was examined by an in situ TdT-mediated dUTP nick-end labeling(TUNEL)method.Results:Activated ERK1/2 was very weakly expressed in the gastric mucosa of normal rats.ERK1/2 was rapidly activated in the gastric mucosa of rats 15 min after WIR stress and the activity reached the maximal after 3.5 h.Pretreatment with PD98059 significantly inhibited ERK1/2 activation,decreased AP-1 and NF-?B activities and TNF-?and IL-1?mRNA expression,and obviously relieved gastric mucosal lesions,accompanied by caspase-3 activation and increased apoptosis.Conclusion:The present results indicate that ERK1/2 activation plays an important role in the development of stress-induced gastric ulcer.

Objective To investigate effects of emergent percutaneous coronary intervention(EPCI)on the prognosis of patients with cardiac arrest(CA)and cardiopulmonary resuscitation after acute myocardial infarction(AMI).Methods Thirty-two patients with CA and carcliopulmonary resuscitation after AMI were retrospectively reviewed and divided into EPCI group(n=12)and eonservative treatment group(n=20).EPCI was used during or after primary eardiopulmonary resuscitation in EPCI group but not in conservative treatment group.Mortality and occurrence rates of severe arrhythmia,of heart failure,of cardiac shock and of severe bleeding in hospital were compared between the two groups.Results The mortality in hospital was 17% in EPCI group and 70% in conservative treatment group,respectively(P＜0.001).Heart failure was developed in 3 patients from EPCI group and 16 patients from conservative treatment group,respectively(P＜0.01).There was no significant difference in severe arrhythmia,cardiac shock and severe bleeding between two groups(P＞0.05).Logistic regression showed that EPCI decreased mortality but the prolonged cardiopulmonary resuscitation(＞20 min)increased the mortality.Conclusion Early EPCI might improve the prognosis of patients with cardiac arrest and cardiopulmonary resuscitation after acute myocardial infarction.

The ergosterols were produced from corn straw hydrolysates fermented by ergosterol yeast,which was obtained from protoplast electrofusion.The effects on the yield of ergosterol were studied in the condition of shaker,such as initial sugar concentration,nitrogen source,pH value and fermentation time.The technical conditions were optimized according to the DPS center-united experimental design principles and the method of response surface analysis with four factors and three levels.The results indicated that the four factors had significant correlation to ergosterol accumulation.The biomass and the ergosterol content could be up to 8.67g/L and 2.37% respectively after cultivated for 32h under optimal technical condition.The structure of ergosterol crystal was characterized by UV,IR and SEM.A new approach of biomass source application was presented.

Objective To compare the effect of right ventricular outflow tract (RVOT) and right ventricular apex (RVA) pacing on ventricular systolic synchrony using gated blood pool SPECT (GBPS).Methods A total of 50 patients implanted with pacemaker due to high degree or complete atria-ventricular block were enrolled in the study. Twenty-three patients were RVOT paced ( Group A, n = 23) and 27 were RVA paced (Group B, n=27). Twenty-four patients with malignancy, normal echocardiographic findings and no history of cardiac diseases were scheduled for pre-chemotherapy evaluation of cardiac structure and function and were enrolled as control group ( Group C, n = 24). All patients underwent GBPS imaging and the values of phase angle (PS), mean phase of each wall, standard deviation (SD) of mean phase of each wall, lateral-septal motion delay of left ventricle ( LV Sep-Lat Delay), septal-right ventricular (RV) delay of LV ( LV Sep-RV Delay) and LV-RV Delay were acquired. The parameters of ventricular systolic synchrony among the three groups were compared using one-way ANOVA. Results The mean phase of LV lateral wall in Groups A and B were significantly higher than that in Group C: Group A (120.50 ±40.58) ms; Group B (103.23±28.34) ms; Group C (84.63 ±22.38) ms (F=7.72, P ＜0.05). There was no significant difference between Groups A and B ( t = 1.30, P ＞ 0.05 ). The mean phase of RV in Group A was significantly larger than those in Groups B and C: Group A ( 137.05 ± 39.27) ms, Group B ( 100.85 ± 23.79) ms,Group C (59. 13 ±30.52) ms (F=35.55, P＜0.05). PS, SD and LV Sep-Lat Delay in Groups A and B were significantly higher than those in Group C: (85.73 ± 12.00)°vs (89.85 ± 15.61 )°vs (58.95 ±9.87)°, (27.68±10.66) ms vs (26.15 ±13.02) ms vs (15.63 ±8.35) ms, (25.06±34.23) ms vs (2. 62 ± 60. 31 ) ms vs ( - 23.66 ± 31.39) ms, F = 41.54,8.55,6.81, all P ＜ 0.01 ), however, there was no significant difference between Groups A and B ( t = 0. 68, 0.68, 1.30, all P ＞ 0.05 ). LV Sep-RV Delay and LV-RV Delay were significantly different among the three groups ( LV Sep-RV Delay: Group A (57.60 ±56.77) ms, Group B (6.36 ±61.88) ms, Group C ( -41.89 ±35.78) ms; LV-RV Delay:Group A (47.36 ±42.59) ms, Group B ( 3.08 ± 38.81 ) ms Group C ( - 26.50 ± 20.99 ) ms, F = 20. 32,25.38, both P ＜ 0.01 ). Conclusion Both RVA and RVOT pacing increase the segmental phases detected by GBPS, causing inter- and intra- ventricular asynchrony compared with patients without pacemakers.

Objective:To establish a human lung squamous carcinoma cell line and to study its biological characteristics. Methods:Lung squamous carcinoma specimens were freshly resected during operation;the tissues were incubated in vitro and the cell line was named CHLH-1.The biological characteristics of the cells were studied by light microscopy,electron microsco- py,chromosome analysis and transplantation experiment.Results:Cells from the specimens of the primary tumor,the CHLC- 1 cell line and the cells from transplanted tumor possessed the characteristics of malignant squamous epithelium under light and electron microscope.The cell growth curve,doubling time and mitotic index were also observed in vitro.Nuclear chromosome analysis revealed that the tumor was a subtriploid with a mode of 60-68 per cell.Tumor nodes were observed under the skin of nude mice by heterogenic transplantation.Conclusion:The characteristics of the established cell line suggest that it is a newly established human squamous carcinoma cell line.