Liver transplantation is the only definitive treatment modality of end stage liver diseases. Demyelinative disease is rarely seen in patients after liver transplantation, but a higher incidence has been noticed recently. The disease is liable to be misdiagnosed as immunosuppressant-induced psychiatric disorders at early stage. Central pontine myelinolysis is more disastrous and it will greatly influence the short-term survival and long-term life quality of the patients after liver transplantation. When it is manifested as peripheral nervous system disorder, the disease usually has a subtle onset, making it difficult for diagnosis and treatment. The causes for various demyelinative diseases should be understood to prevent it in patients after liver transplantation. The specific mechanism for demyelinative disease after liver transplantation remains unclear. We introduce the possible causes of common demyelinative diseases, hoping to provide reference for prevention and treatment of such conditions after liver transplantation.

Objective To observe the influence of serum containing Rhizoma Chuanxiong on the proliferation and differentiation of mouse embryos stem cells(ESCs).Methods Mouse ESCs were co-cultured with serum containing Rhizoma Chuanxiong.The proliferation of ESCs was detected by CCK-8 method.The expression level of specific gene beta-myosin heavy chain (β-MHC) in cardiac myoblasts was detected by reverse transcription-polymerase chain reaction (RT-PCR).Results Compared with the blank rat serum group and the blank fetal bovine serum group,the differences of activities of ESCs in serum containing Rhizoma Chuanxiong group were in significant(P ＞0.05) and the expression level of specific gene β-MHC in cardiac myoblasts was increased (P ＜ 0.05).Conclusion Serum containing Rhizoma Chuanxiong can promote the differentiation of ESCs into cardiac myoblasts.

Objective To explore a suitable method to estimate the quantity of removing skin in double-fold eyelid operation.Methods Upper eyelid skin gently pulled towards eyebrows was flattened with the eyelash being slightly curved,the double-fold eyelid line that was parallelled to blepharitis was marked at 6-7 mm far from blepharitis.Then opening eyelid and looking upwards,the suprapalpebral fold was formed,and several dots were marked on the fold margin. After slightly closing eyelid the suprapalpebral fold margin line was formed through connecting the dots.The skin between the two line was the quantity to remove in operation.If the two line overlaped,removing skin did not need.Results A total of 89 cases were enrolled in this study.The patients were followed up from three months to one year,and the results showed that 67 cases were satisfactory,22 cases were basically satisfatory,and no one was not satisfactory.Conclusions The designing method for removing skin according to double-fold eyelid line and suprapalpebral fold margin line is a good method to estimate the quantity of removing skin in double-fold eyelid operation.

Objective To study the immunosuppressive mechanism of alkaloid sinomenine (SIN) by observing the effects of SIN on the proliferation and intracellular protein expression levels of nuclear factor of activated T cells (NF-AT) and interferon-gamma (IFN-γ) in CD4+ T lymphocytes of human periphery blood. Methods CD4+ T lymphocytes were isolated from PBMC suspensions with immunomagnetic beads and divided into five groups to culture. (1) Negative control group: no medicine was added to cell culture medium; (2) Positive control group: CsA solution (final concentration: 50ng/ml) was added to cell culture media; (3) Low-concentration SIN group (L-SIN): low-concentration SIN solution (final concentration: 10 μmol/L) was added to cell culture media; (4) Middle-concentration SIN group (M-SIN): middle-concentration SIN solution (final concentration: 200 μmol/L) was added to cell culture media; (5) High-concentration SIN group (H-SIN): high-concentration SIN solution (final concentration: 1000 tmol/L) was added to cell culture media. The proliferations of CD4+ T lymphocytes were observed. Western blotting was performed to detect the protein expression levels of NF-AT. FCM was used to determine the levels of IFN-γ. Results Compared with negative control group, the cell proliferation was significantly inhibited in H- and M-SIN groups (P＜0. 01 ). SIN concentration-dependently inhibited the protein expression levels of NF-AT and IFN-γ in CD4+ T lymphocytes of human periphery blood (P＜0.01). The protein expression levels of NF-AT and IFN-γ were lowest in positive control group. There was a close negative correlation between intracellular levels of NF-AT and cell proliferation inhibition ratio in CD4+ T lymphocytes of human periphery blood (rs = - 0. 969, P = 0. 000). Conclusion SIN can inhibit the protein expression of NF-AT and IFN-γ in CD4+ T lymphocytes of human periphery blood probably by decreasing protein levels of NF-AT to inhibit the activity and proliferation of CD4+ T lymphocytes.

Objective: To investigate the expression changes of Bcl-2-associated athanogene 1(BAG-1) and its regulatory effect on the glucocorticoid receptor(GR) activity in rat alveolar macrophages in conditions of cell inflammation and glucocorticoid therapy.Methods: The expression changes of BAG-1 were detected by Western blot after lipopolysaccharide(LPS) and Dexamethasone(Dex) treatment of rat alveolar macrophages(AMs),the interaction between BAG-1 and GR determined by immune coprecipitation experiment,and the transcriptional activation of GR measured by relative luciferase activity assay.Results:After LPS and Dex treatment,the expression of BAG-1L in total protein increased but that of BAG-1S remained changed,BAG-1L rather than BAG-1S was detected in nuclear protein and its expression increased gradually within 24 hours,the interaction between BAG-1L and GR was observed in nucleoli,and the transcriptional activation of GR decreased,with a negative correlation between BAG-1L expression and GR activity.Conclusion:LPS and Dex acting on rat alveolar macrophages,the expression of BAG-1L increases,which,coupled with GR,translocates into nucleoli and inhibits GR activity.This might be the important mechanism that underlies glucocorticoid resistance in inflammation.

OBJECTIVETo introduce practical diagnostic criterion of blood stasis syndrome (BSS), and to evaluate its reliability and validity.

METHODSBy referring to three diagnostic criteria of BSS [practical diagnostic criterion of BSS (criterion A), diagnostic criterion of BSS in 1986 (criterion B), Consensus of Integrative Medicine on BSS Diagnosis in 2011 (criterion C)], 712 patients from different departments of Xiyuan Hospital were recruited. The reliability of criterion A and its consistency with the other two criteria were assessed using Kappa coefficient. A Bayesian approach was also employed to assess the sensitivity and specificity of criterion A.

RESULTSAccording to the consistency check, criterion A presented good consistency when used by different researchers (the diagnostic accordance rate was 91. 96%, Kappa =0. 82, P <0.001). Meanwhile, there was an acceptable diagnostic consistency among the three diagnostic criteria. Bayesian estimation suggested that criterion A had higher sensitivity but similar specificity, as compared with criterion B or criterion C. Compared with criterion B [the median of sensitivity and specificity were 0. 762 (95% Cl: 0. 731 -0. 790) and 0. 902 (95% Cl: 0. 858 -0. 936) respectively, the median of sensitivity and specificity of criterion A were 0. 911 (95% CI: 0. 888 - 0. 930) and 0. 875 (95% CI: 0. 826 - 0. 915) respectively. Estimating the difference between criterion A and B, the median of sensitivity and specificity were 0. 149 (95% CI: 0. 112 -0.184) and -0. 026 (95% CI:-0. 085 -0. 033) respectively. Compared with criterion C [the median of sensitivity and specificity were 0. 831 (95% Cl: 0. 804 -0. 857) and 0. 892 (95% CI: 0. 848 - 0. 926) respectively], the median of sensitivity and specificity of criterion A were 0. 912 (95% CI: 0. 889 -0. 932) and 0. 880 (95%CI: 0. 833 - 0.919) respectively. Estimating the difference between criterion A and C, the median of sensitivity and specificity were 0. 081 (95% CI: 0.047 - 0.114) and -0.011 (95%CI: -0.070 -0.046) respectively.

CONCLUSIONCompared with criterion B and C, criterion A not only had better reliability, but also could significantly improve the sensitivity without obviously lowering the specificity.

This study aimed to analyze the etiology of the encephalitis outbreak in Longyan, Fujian Province, China in 2010, in order to provide valuable information for this prevention and control of this disease. Pathogens were confirmed from cerebrospinal fluid samples with fluorescent RT-PCR, virus isolation (RD cells), and neutralization tests. Then, the VP1 fragments or whole genome nucleotide sequences were determined for four virus strains using PCR. Homology was assessed using the MegAlign software, and a phylogenetic evolutionary tree was drawn using Mega 4.0 software. The results confirmed that the etiology of the outbreak was the ECHO6 intestinal virus, and the nucleotide sequence of the VP1 segment indicated that the C2 subtype was responsible. The genome sequence consisted of 7407 nucleotides, and resembled the genome of other ECHO and CoxB viruses with homology levels of 78.5%-87.3%. The encephalitis outbreak in Longyan in 2010 was caused by the ECHO6 C2 subtype intestinal virus, and its complete genome sequence length is similar to the standard strain (U16283) with a sequence homology of 80.4%.
Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; Echovirus 6, Human ; classification ; genetics ; isolation & purification ; Echovirus Infections ; epidemiology ; virology ; Encephalitis ; epidemiology ; virology ; Female ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny

Objectives: To study the correlation between plasma matrix metalloproteinase 9(MMP 9) concentration and acute lung injury following cardiopulmonary bypass(CPB). Methods: Human plasma was obtained after informed consent from twenty patients undergoing CPB. Plasma was collected at the beginning of CPB, 5 minutes after the initiation of CPB, at the termination of CPB, 1 hour after the termination of CPB and 6 hours after the termination of CPB. All samples were analyzed by standard enzyme linked immunosorbent assay (ELISA). A aDO 2 and respiratory index (RI) was measured at the termination, 1 hour and 6 hours after termination of CPB. The cross clamp times, CPB times and the time to extubation was recorded. Data were expressed as means ?SE and assessed by analysis of variance (ANOVA).The regression analysis was utilized to define the correlations of variables measured( A aDO 2 ,RI, cross clamp times, CPB times and the time to extubation ) at the end of CPB. Results: Plasma MMP 9 concentration was significantly increased at the end of CPB (430.6?50)?g/L( P

Objective To study the effects of using 25 kDa linear and branched PEI to transfer plasmid DNA pEGFP -C1 (pDNA ,encoding the enhanced green fluorescent protein reporter gene ) to the basilar membrane of the C57BL/6 mice cochlea in vitro .Methods L -PEI/pDNA and B -PEI/pDNA polyplexes were generated in 0 .1M phosphate buffer solution (PBS) or 5% glucose solution .Polyplexes were characterized by transmission electron mi-croscopy .Agarose gel retardation assay was used to determine the plasmid binding ability of L -PEI and B -PEI . The toxicity was investigated by MTT assay .The transfection was firstly evaluated in 293T cell line ,and then the appropriate amount of PEI and plasmid were applied for cochlear explant transfection of P4 mice pups .Results Un-der the same condition ,B -PEI had better transfection efficiency than L -PEI ,but its toxicity was also higher . When generated in PBS ,the polyplexes had lower toxicity than in glucose solution .L -PEI-pDNA nanoparticles could transfect the spiral limbus fibrocytes ,some spiral ganglion neurons and supporting cells ,but the efficiency was low .Conclusion L -PEI could be used as the non -viral vector for the transfection of the cultured basilar mem-brane of P4 mice pups ,but it should be modified to reach higher efficiency .

Background The proliferation and migration of vascular endothelial cells is a primary link during angiogenesis.Studies showed that heat shock protein B6 (HspB6) promotes the secretion of multiple angiogenesis-related factors and therefore leads to neovascularization.Understanding the effects of neutralizing HspB6 antibody on the biological behavior of human choroidal vascular endothelial cells has an important significance in the target treatment of choroidal neovacularization diseases.Objective This study was to address the role and mechanism of neutralizing HspB6 antibody in tube formation of human choroidal vascular endothelial cells.Methods Human choroidal vascular endothelial cell line was normally cultured and harvested for total RNA extraction.Expressions of HspB6 mRNA and protein in human choroidal vascular endothelial cells were detected by reverse transcription PCR (RT-PCR) and flow cytometry (FCM).The cells were seeded on 96-well plate covered with matrigel at the density of 2×104/hole.Then the neutralizing HspB6 antibody at the concentration of 100 μg/Land 500 μg/L was added into the medium respectively,and the control cells were set without the addition of HspB6 antibody.The number of capillary tubes was calculated 12 hours after culture by three-dimensional matrigel assay.In addition,0,50,100,500 μg/L of neutralizing HspB6 antibody were added into the cell medium separately for 24hours,cell counting kit-8 (CCK-8) method was employed to assay the inhibitory rate(IR) of the cells.Transwell test was used to count the cell number across chamber membrane for the evaluation of migration ability of the cells.The apoptosis of the cells was assayed by FCM.Results Both HspB6 mRNA and protein were expressed on human choroidal vascular endothelial cells.The number of capillary tube formation of human choroidal vascular endothelial cells was (67.25±5.75),(60.39±6.41) and (39.76±10.73) /field in the 0,100 and 500 μg/L neutralizing HspB6 antibody groups,with significant difference among them (F =10.210,P =0.012),and the tube number was significantly less in the 500 μg/L neutralizing HspB6 antibody group compared with 0 μg/L neutralizing HspB6 group (P =0.005).The IR of neutralizing HspB6 antibody to the cellular proliferation and migration was enhanced with the increases of concentration and time lapse(Fconcentration =7.485,P =0.002 ; Ftime =16.684,P =0.001).The number of the cells through Transwell chamber membrane was 14.0 ± 2.5,11.1 ± 0.8,6.6 ± 0.1,6.7 ± 0.2 in the 0,50,100,500 μg/L neutralizing HspB6 antibody group respectively,and that in the 100 μg/L and 500 μg/L neutralizing HspB6 antibody group was lessened in comparison with the 0 μg/L neutralizing HspB6 antibody group(both at P=0.000).The apoptosis rate of the cells was (22.73 ± 2.53)％ in the neutralizing HspB6 antibody group,which was significantly lower than (13.33±2.08) ％ of the control group (t=4.967,P=0.008).Conclusions Neutralizing HspB6 antibody inhibits capillary tube formation of human choroidal endothelial cells in vitro in dose-and timedependent manner,probably through suppressing the proliferation and migration and promoting the apoptosis of choroidal endothelial cells.