Objective To investigate the safety of left atrial appendage occlusion by silk thread ligation during open heart op‐eration in patients with rheumatic atrial fibrillation ,and to evaluate its effectiveness for prevention of cerebral embolism .Methods From April 2012 to March 2014 ,129 patients with rheumatic atrial fibrillation were undergone mitral valve replacement and left at‐rial appendage occlusion by ligation using two silk threads from the outside of the heart (ligation group) .The indexes related to the operation ,postoperative complications incidence ,and cerebral embolism incidence during the follow‐up period of ligation group were compared with the indexes of another 129 patients without ligation of left atrial appendage over the same period (control group) . Results The operation time ,the cardiopulmonary bypass time ,the clamp time ,the intensive care unit stay time ,the postoperative hospitalization time in ligation group were (235 ± 50)min ,(88 ± 24)min ,(57 ± 16)min ,(26 .5 ± 9 .3)h and (12 .4 ± 7 .5)d respective‐ly ,and significant difference was not found compared with control group (P>0 .05) .The thoracotomy for hemostasis(1 cases) ,low cardiac output syndrome(2 cases) ,acute renal failure(2 cases) ,pulmonary infection(3 cases) ,sternal wound dehiscence(2 cases) and other complications in ligation group had no significant difference ,compared with control group(P>0 .05);2 cases died in liga‐tion group ,3 patients died in control group ,the differences had no statistical significance(P>0 .05) .No cerebral embolism occurred in ligation group with 127 patients following‐up (23 .6 ± 11 .3) months ,but 5 patients suffered from cerebral embolism in control group with 126 patients following‐up (22 .9 ± 12 .1) months ,the difference had statistical significance(P<0 .05) .Conclusion Left atrial appendage occlusion by silk thread ligation during open heart operation in patients with rheumatic atrial fibrillation is simple and safe ,can reduce cerebral embolism incidence .

Objective To observe the PML protein expression of hepatocellar carcinoma tissue and cells lines and As 2 O3 regulate its expression .Methods Immunohistochemistry was used to examine the PML protein expression of hepatocellar carcinoma tissue . Western blot analysis were used to observe PML protein expression of hepatocellar carcinoma tissue of 12 cases ,5 hepatocellar car-cinoma cell lines ,such as HuH7 ,HepG2 ,Hep3B ,SMMC-7721 ,MHC97H .Western blot analysis was used to detected the PML pro-tein expression of these hepatocellar carcinoma cell lines after 72-96 h treated with 0 .25 μg/mL of As2 O3 .Results Immunohisch-enmical staining showed that the PML protein was expressed in both cytoplasm and nucleus ,did not well-distributed in hepatocellar carcinoma cells .There was no significant differences of PML protein expressed among differently differentiated stages of hepatocel-lar carcinoma cells .Western blot analysis found that hepatocellar carcinoma tissues of 12 cases with hepatocellar carcinoma ex-pressed PML protein ,and there was significant difference of PML protein expressed among 12 cases suffer with hepatocellar carci-noma .hepatocellar carcinoma cell lines ,such as HuH7 ,HepG2 ,Hep3B ,SMMC-7721 and MHC97H all expressed PML protein ,and there was little difference of PML protein expressed among hepatocellar carcinoma cell lines .The PML protein expression of HuH7 ,HepG2 ,Hep3B ,SMMC-7721 and MHC97H cell after 72-96 h treated with 0 .25 μg/mL of As2O3 significant decreased . Conclusion Hepatocellar carcinoma tissue and cells may express PML protein ,and As2 O3 may regulate this protein expression as well .PML protein may be the target molecule of As2 O3 treating HCC .

Background. Our previous studies indicated that the increased arginine vasopressin(AVP) in ischemic brain regions of gerbils could exacerbate the ischemic brain edema. This experiments is further clarify the relation between AVP and cerebral ischemia at the molecular level. Methods. The contents of AVP, AVP mRNA, AVP immunoreactive(ir) neurons in supraoptic nucleus(SON)and paraventricular nucleus(PVN) after cerebral ischemia and reperfusion were respectively determined by radioim-munoassay(RIA), immunocytochemistry( Ⅱ C), situ hybridization and computed image pattem analysis. Results. The contents of AVP in SON, PVN were increased, and the AVP ir positive neurons in SON and PVN were also significantly increased as compared with the controls after ischemia and reperfusion. And there were very light staining of AVP ir positive neurons in the other brain areas such as suprachiasmatic nucleus (SC) and periven-tricular hypothalamic nucleus (PE), but these have no significant changes as compared with the controls. During dif-ferent periods of cerebral ischemia (30～ 120 min) and reperfusion (30 min), AVP mRNA expression in SON and PVN were more markedly increased than the controls. Condusions. The transcription of AVP gene elevated, then promoting synthesis and release of AVP in SON,PVN. Under the specific condition of cerebral ischemia and repeffusion, the activity and contents of central AVP in-creased abnormally is one of the important factors which causes ischemia brain damage.

This paper was aim to screen microorganisms with attenualed efficiency for Chinese medicine containing aristolochic acid A by liquid-state fermentation. Twelve Chinese medicine were detected by UPLC and aristolochic acid A was only founded in four species of Aristolochia, those were Caulis Aristolochiae Manshuriensis, Aristolochiae Radix, Aistolochia Contorta Bunge and Herba Aristolochiae Mollissima,but not in the others. With the four Chinese medicine containing aristolochic acid A as raw material, ten microorganisms were tested, and the content of aristolochic acid A was detected by UPLC. The results showed that one microorganism can decrease content of aristolochic acid A in all those four Chinese medicine.
Aristolochic Acids ; analysis ; metabolism ; Bacteria ; metabolism ; Biotransformation ; Drugs, Chinese Herbal ; analysis ; metabolism ; Fungi ; metabolism ; Plants, Medicinal ; chemistry ; microbiology

At present time, the widely used hepatitis B virus( HBV) vaccines consist of only the small hepatitis B surface antigen expressed in yeast or CHO cells. The frequency of non-responders to these vaccines has increased the demand for a more immunogenic vaccine. Some studies have suggested that the addition of preS region to the vaccine will improve its efficacy. However, the large protein (L) containing the whole preS region can not be effectively expressed in vitro. To overcome this problem, two chimeric contructs, SS1, surface gene containing preS1 region at C-terminus and SS2, surface gene containing preS2 region at C-terminus, were constructed and effectively expressed in our previous studies. Here we further constructed an expression vector containing both SS1 and SS2 expression cassettes by separation and ligation the SS2 cassette to a linearized SS1 expression vector pAO815-SS1. The recombinant vector was transformed into Pichia pastoris GS115 by electroporation. A high-level expression strain (GS115-SS1S2) was established by primary screening for His+ transformants and further analysis for induction products. ELISA results demonstrated that the expressed protein had S, preS1 and preS2 antigenicities simultaneously. Western blotting showed that the product can bind to all of the three antibodies, anti-S, anti-preS1 and anti-preS2. The expression protein was present in the form of particles of 20-35 nm diameter and the yield of recombinant particles reached 300-600 mg/L by fermentation. The SS1 and SS2 polypeptides kept intact in purified particles, suggesting that the stability of preS region has been significantly improved.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Epitopes ; genetics ; immunology ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B Vaccines ; immunology ; Peptide Fragments ; genetics ; immunology ; Pichia ; genetics ; Protein Precursors ; genetics ; immunology ; Vaccines, Synthetic ; immunology

Objective To investigate the effects of different dialysates on expression of protein kinase C-δ (PKCδ) and apoptosis of U937 cell line. Methods Different dialysates were added into culture fluid with U937 cell line at exponential phase of growth, and groups were divided: fluid A+fluid B group (dialysate A+dialysate B), fluid A+fluid B+rottlerin (PKCδ specific inhibitor)group, fluid A+powder B group (dialysate A+powder B) and fluid A+powder B + rottlerin group. Besides, blank control group and normal control group were established. Cells were harvested 24 h and 48 h after treatment, morphological changes were observed by Hoechst33258 fluorescence staining, cell apoptosis was measured by Annexin-V-FITC/PI double staining, and expression of PKCδ mRNA and protein was detected by RT-PCR and Western blotting, respectively. Results Cell apoptosis significantly increased in fluid A+powder B group, with typical morphology of apoptosis. After treatment for 24 h and 48 h, cell apoptosis rates in fluid A+powder B group were significantly higher than those at corresponding time points in blank control group , normal control group and fluid A+powder B+rottlerin group (P<0.05). Compared with normal control group, blank control group and fluid A+powder B+rottlerin group, the expression of PKCδ mRNA and protein of U937 cells in fluid A+powder B group were significantly increased (P<0.05). There was no significant difference in cell apoptosis rates and expression of PKCδ mRNA and protein between fluid A+fluid B group and blank control group, normal control group and fluid A+fluid B+rottlerin group (P＞0.05). Conclusion Fluid A+powder B can significantly increase apoptosis of U937 cell line, the mechanism of which may be associated with the up-regulation of expression of PKCδ. Compared with fluid A+powder B, fluid A+fluid B is superior in reducing apoptosis of peripheral blood monouclear cells.

The preparation process and immunogenicity of a novel hepatitis B vaccine containing preS1, preS2 and S epitopes were investigated in this study. A Pichia pastoris stain GS115-SS1S2 harbouring two chimeric HBsAg gene constructs, SS1 and SS2 was cultivated by high-density fermentation. 300-600 mg/L of the expression level was achieved through 48-72 h methanol induction. SSIS2 antigen was extracted and purified by silica adsorption, HIC and SEC to 99% purity from the harvested cells. 82 mg purified antigen could be achieved from one liter of fermentation culture. The immunogenicity of the purified antigen was evaluated in NIH mice. Three groups of female NIH mice, 14-16 g in weight, were injected once intraperitoneally with 2.5, 0.625, 0.156 microg of each of the two vaccines: SS1S2 or a commercially available S vaccine. Part of the mice were bled in 30 days after injection to compare the ED50 of the two vaccines. For the SSIS2 vaccine, the ED50 is 0.46, 0.29 and 0.84 microg respectively for the preS1, preS2 and S antigens. For the S vaccine, the ED50 is 0.99 microg for the S antigen. Another part of the mice were bleed in 7 or 14 days to detect preS1, preS2 and S antibodies. Higher ratios of mice were seroconverted for preS1 and preS2 antibodies as compared to the S antibody in these two time points. These results suggest that the SS1S2 vaccine may be more immunogenic than the conventional S vaccines.
Animals ; Epitopes ; immunology ; Female ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B Vaccines ; biosynthesis ; immunology ; Mice ; Pichia ; genetics ; metabolism ; Protein Engineering ; Protein Precursors ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Viral Envelope Proteins ; immunology

This article introduces a new-type anti-rotation reduction internal fixator, which can be applied in various spine fractures and dislocations in order to shorten the operation time, to raise reduction effect, and to reduce the complications such as the loss of reduction, broken nail, broken rod etc. Biomechanical tests and clinical applications have proved that the internal fixator has the features of a short operation time, a definite fixation and few complications.
Bone Nails ; Bone Plates ; Equipment Design ; Fracture Fixation, Internal ; instrumentation ; methods ; Humans ; Internal Fixators ; Spinal Fractures ; surgery

OBJECTIVETo study the effect of concentrations of glucose on the initial adherence of Streptococcus mutans (S. mutans) to saliva-coated hydroxyapatite (SHA), and to compare the initial adherence of S. mutans from caries-active group with that of S. mutans from caries-free group.

METHODSEach 10 clinical isolates of S. mutans from caries-active and caries-free subjects were used in this study. And S. mutans UA159 was also included in this experiment. SHA was used to simulate tooth surface in oral cavity. S. mutans clinical isolates and strain UA159 were cultured in TPY liquid medium containing 3H-TdR in the same radioactive concentration and glucose in 0.2%, 1.0%, 5.0% concentration. Then grown cells were harvested to produce a suspension. SHA and radiolabelled bacterial suspension (A550(nm) = 0.52) were mixed for 90 minutes, samples were assayed by using liquid scintillation counter, and binding abilities of strains were evaluated by the count per minute (CPM).

RESULTSThe initial adherence ability of S. mutans from caries-active group was higher than that of S. mutans from caries-free group (P < 0.05). And the initial adherence ability of S. mutans cultured in different concentration of glucose was also significantly different (P < 0.05), 5.0% glucose group had the highest adherence ability, and 0.2% glucose group had the lowest adherenceability.

CONCLUSION(1)Difference of the initial adherence of S. mutans might relate to difference of carious experiences; (2) Glucose may play an important role in S. mutans initial adherence, to some extent, S. mutans cultured in the higher concentration of glucose has the higher initial adherence property.

OBJECTIVETo study the pattern of polymorphism expression of heat-shock protein 70 (HSP70) family in A549 cell line treated with different concentrations of benzo(a)pyrene (BaP) and its probable biological effect.

METHODTwo-dimensional polyacrylamide gel electrophoresis (2D-PAGE) was used for the HSP70 expression analysis.

RESULTS2D-PAGE showed that when A549 cells were exposed to different concentrations of BaP (0.1, 1.0, 5.0, 10.0 micromol/L) for 24, 48 h respectively, the HSP72 in A549 gradually declined as BaP concentrations increased [the integral OD (IOD)] for 24 h were: 150.36 +/- 26.03, 98.57 +/- 13.34, 64.92 +/- 15.03, 34.65 +/- 19.10, 32.92 +/- 18.71 respectively, for 48 h: 126.85 +/- 17.41, 106.19 +/- 15.32, 73.64 +/- 21.02, 35.18 +/- 11.95, 16.27 +/- 9.35 respectively), while the IOD of HSP73 did not show any remarkable change (24 h: 102.29 +/- 21.24, 87.71 +/- 18.70, 71.19 +/- 14.08, 71.87 +/- 15.16, 72.78 +/- 17.31 respectively; 48 h: 86.66 +/- 16.86, 75.67 +/- 10.61, 66.83 +/- 12.63, 67.29 +/- 10.26, 91.37 +/- 13.68 respectively).

CONCLUSIONBaP can inhibit HSP72 expression and with certain dose-effect relationship, but cannot affect HSP73 expression.