Objective To construct the recombinant plasmid containing human filaggrin gene,purify and identify the immunoreactivity of the recombinant protein,and establish the indirect ELISA to detect AFA for diagnosis of RA.Methods The constructed plasmids were transformed into E. Coli Rosettagami(DE3).This fusion protein was purified by NAT chromatography.ELISA coated with the fusion protein Was established to detect the AFA in serum of patients,which included 114 cases of RA,56 cases of SLE,32 cases of OA and 40 cases of normal controls. The correlation between the results of AFA and anti-CCP in RA group were compared. Results 321 bp fragment of filaggrin gene was amplified and the recombinant expression vector pET-28a( + )-filaggrin was constructed. The sequence of filaggrin gene was the same as the sequence reported in the literatures. The Rosetta-gami (DE3) strains of E. Coli with recombinant vector showed high level of filaggrin protein after induction. The SDS-PAGE showed that the plasmid expressed the filaggrin fusion protein with molecule weight of 14 000 Da. The expression protein could be purified by Ni-NAT with activity. The absorbance value of AFA in RA group was 0.473 ±0. 248 while they were 0. 160 0. 088, 0. 121±0. 070, 0.050 0. ±018 in SLE, OA and normal groups respectively. There were significant differences of absorbance values of AFA between RA and SLE, OA, control group (t = 12.004, 14. 464, 18.078, P<0. 01, respectively). The positivities of anti-filaggrin in RA, SLE and OA were 48.2%, 5.4% and 3. 1% respectively. The positivities of AFA were significantly different between RA, OA and normal control groups (x~2 = 67. 088, P < 0. 01). There was positive correlation of results between AFA and anti-CCP antibody (r = 0.42, P < 0. 05 ) . The consistency rate of results between AFA and anti-CCP was 70. 1%. Anti-CCP was negative in 10 out of 114 patients with AFA positive. AFA can be used to diagnose RA with sensitivity of 48. 2% , specificity of 96.9% , positive predictive value of 93. 2% and negative predictive values of 67. 9% . Conclusions The purified human filaggrin fusion protein is successfully purified. The indirect ELISA method based on the recombinant protein shows good sensitivity and specificity. Joint detection with AFA and anti-CCP can improve the positive rate of detection.

ObjectiveTo explore the application and significance of assisted laparoscopic hepatectomy (ALH) in living-donor-hepatectomy.MethodsWe successfully performed 7 cases of ALH of right hepatectomy on living donor from 30/5/2011 to 1/9/2011.ResultsThe donors recovered well with ratio of remnant lver:32.10％ ～38.31 ％,good liver fuction,little pain and no surgical complications.All the wound sutured intracuteneously was taken out stitches 7 days after operation and healed perfectly.Liver function got normal 2 weeks after operation.Conclusions ALH,which gives the consideration to both the minimal invasion of laparoscopic surgery and safe of open surgery,can be applied safely in hepatectomy of living donor and highly acceptible for donor and receptor.

Objective This study was to explore deaf people's needs for a smooth communication with doctors and/or nurses during health consultation in the outpatient department or when being hospitalized and supply reference for compilation of sign language textbooks.Methods Purposive sampling was used and semi-structured interviews were conducted.Six deaf participants who met the inclusion and exclusion criteria were interviewed.It lased for 20 to 40 minutes for each interview.Interviews were recorded digitally and then transcribed and validated.Transcripts were analyzed using a method of thematic analysis.Results Three themes were identified,i.e.needs for sign language interpreters,needs for information and knowledge.and needs for psychosocial and emotional support.Needs for sign language interpreters meant that doctors and nurses were able to communicate with them using simple sign language instead of terminology.The required information and knowledge covered three aspects,i.e.health condition,pharmaceutical therapy,and self-care activity.Conclusions There may exist serious communicative barriers for deaf people during their health consultation in the outpatient department or when being hospitalized.There is a need for healthcare professionals who are competent in sign language to establish the health delivery environment with minimal barriers.

Objective To retrospectively analyze and conclude the characteristics of imaging appearances of solid psedopapillary tumor of pancreas.Methods Among 7 cases with pathologically proved solid psedopapillary tumor of pancreas,6 cases underwent CT examinations of upper abdomen preoperatively,and the rest One had MRI examination.The mean age of these 7 cases(all female)was 30.7 years(range,14—44 years).Results The tumors were usually quite large in the largest diameter ranged from 2.8 to 15.9cm(mean largest diameter,7.9 cm);Tumors were all well demarcated,and 5 of them were of capsule on CT or MR imaging.All tumors were well-encapsulated on pathologic specimens, except for the capsule of 1 tumor was partially invaded;In 6 cases underwent CT examination,scattered, punctate and linear calcification were noted in the capsule of 2 tumors and the rim of another one;Except for 1 tumor was almost solid,the other 6 tumors contained both solid and cystic components;Scattered sheets of high attenuation shown in the cystic or solid parts on CT imaging in several cases and the high signal intensity on T_1-weighted MR imaging signified the possibility of bleeding in tumors,which then was testified by pathologic evaluation.Conclusion The solid psedopapillary tumor of pancreas has comparatively characteristic clinical and imaging features.

OBJECTIVETo study the expression ratio of AML1-ETO9a (AE9a) isoform in t(8;21) acute myeloid leukemia (AML) and its clinical significance.

METHODSBone marrow samples from 44 newly diagnosed t(8;21) AML patients co-expressed AE9a and AE were screened by RT-PCR. The alteration of the AE9a expression ratio was monitored during follow-up by using quantitative real-time RT-PCR (qPCR).

RESULTSThe expression level of AE9a was markedly lower than that of AE in these patients. There was a positive correlation between the expression level of AE9a and AE in most of bone marrow samples. The transcript level of both AE9a and AE was decreased in the 44 patients after one course of standard chemotherapy, but the percentage of AE9a expression level was increased in comparison with that before treatment (P < 0.05). After one course of standard chemotherapy treatment, the percentage of AE9a in incomplete remission (ICR) patients was significantly higher than that in CR patients (P < 0.05). Relapsed patients had a higher AE9a ratio than the unrelapsed patients (P < 0.05). During the remission, the percentage of AE9a in 11/17 relapsed patients obviously elevated even while the expression of AE fusion gene at low level.

CONCLUSIONSAE9a and AE co-expressed in most of AML patients with t(8;21) translocation. The expression level of AE9a was lower than that of AE, and there is a positive correlation between the expression level of these two isoforms. The sensitivity of AE9a gene to the standard chemotherapy is less than that of the AE fusion gene. Monitoring the AE9a to AE ratio during the CR can predict the early relapse of the disease compared to monitoring the AE alone.

Adolescent ; Adult ; Child ; Chromosomes, Human, Pair 21 ; Chromosomes, Human, Pair 8 ; Core Binding Factor Alpha 2 Subunit ; genetics ; Female ; Gene Expression ; Humans ; Leukemia, Myeloid, Acute ; genetics ; pathology ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; Protein Isoforms ; genetics ; RUNX1 Translocation Partner 1 Protein ; Translocation, Genetic ; Young Adult

Objective To explore the clinical efficacy and safety of hyper-early embolotherapy in treatment of intracranial ruptured aneurysm.Methods A retrospective analysis was made on 33 patients with intracranial ruptured aneurysm.Preoperative Hunt-Hess grade:grade Ⅰ-Ⅱ in 16 patients,gradeⅢin 5 patients,grade Ⅳ in 9 patients,grade Ⅴ in 3 patients.All patients were confirmed with subarachnoid hemorrhage (SAH) by angiography and then underwent embolization under general anesthesia by detachable coils within 6 h from onset.Results After operation,25 patients (75.8％) recovered well,4 patients (12.1％) were with mild disability with paralysis and aphasia,4 patients (12.1％) were dead (1 patient for intraoperative aneurysm rupture,1 patient for postoperative pneumonia,1 patient for infection of hematoma at puncture site and 1 patient for postoperative gastrointestinal bleeding).Followed up 1-6 months,no rebleeding occurred.Conclusions Hyper-early embolotherapy could avoid rebleeding of the aneurysm,and relieve the vasespasm,without increasing the intra-operative rebleeding rate.Moreover hyper-early embolotherapy could greatly decrease the mortality of poor-grade SAH patients.

This article was aimed to study immunomodulatory effect of chemical split fractions ofMori Cortex, in order to initially explain effective parts that played a role in immunomodulatory effect ofMori Cortex. The carbon clearance test, serum hemolysin test, E-rosette test, and lymphocyte transformation test were carried out to explore influence of these chemical split fractions ofMori Cortex on immune organs, nonspecific immunity, humoral immunity and cellular immunity. The results showed that in the carbon clearance test, 50% ethanol fraction markedly reduced the thymus index (P<0.01) and the correction indexα (P<0.05). In hemolysin test, the half value hemolysis (HC50) was improved by 30% ethanol fraction and fatty oil fraction (P<0.05). Besides, in the E-rosette test, the E-rosette ration was increased in the 30% ethanol fraction group (P<0.05). In the lymphocyte transformation test, the 30% ethanol fraction can promote the thymus and spleen lymphocytes proliferation (P<0.05 orP<0.01), while the 50% ethanol fraction inhibited the proliferation (P<0.05 orP<0.01). It was concluded that the 30% ethanol fraction can boost both the humoral immunity and cellular immunity; the 50% ethanol fraction can induce the growth of thymus with a suppressive effect on nonspecific immunity and cellular immunity; the fatty oil fraction can improve humoral immunity.

Objective To investigate the effect of remote ischemic preconditioning on nuclear factor erythroid-2 related factor 2 (Nrf2) signaling in the liver of aged rats after warm ischemia reperfusion injury.Methods A 70％ hepatic warm ischemia-reperfusion model was established in aged rats (15-16 months old) (1 hour after ischemia and 2 hours after reperfusion).12 male Sprague-Dawley rats were divided into two groups:pretreatment group and control group.The level of serum alanine aminotransferase (ALT) was detected after operation and the liver tissues were harvested for the determination of malondialdehyde (MDA) content and the activity of antioxidant enzyme-superoxide dismutase (SOD).The pathological changes of liver were observed.The change of Nrf2 protein expression in liver tissue was examined by Western blot.Results Serum ALT and MDA in the pretreatment group were significantly lower than those in the control group.The liver pathological damage of pretreatment group rats were lighter than the control group (P ＜ 0.05).Compared with the control group,the Nrf2 protein expression and the activity of SOD increased in the liver of pretreatment group (P ＜ 0.05).Conclusions Remote ischemic preconditioning can reduce 70％ hepatic ischemia-reperfusion injury in aged rats,and its mechanism may be related to its activation of Nrf2 signaling pathway.

OBJECTIVETo investigate of the regulatory effect of Rho-kinase pathway activation on angiotensin II (Ang II)-induced contraction of human airway smooth muscle cells (HASMCs) in vitro.

METHODSCultured primary HASMCs were divided into control group, AngII group, AngII + irbesartan group and AngII + Y-27632 group with corresponding treatment. AngII-induced contraction of HASMCs was evaluated using collagen gel lattices and observed morphologically using immunofluorescence assay. Western Blotting was significantly performed to examine the protein expression of Rho-kinase signal pathway.

RESULTSAngII-induced HASMC contraction was inhibited by treatments with irbesartan and Y-27632 as shown by gel contraction assay (P<0.001). Y-27632 treatment produced a stronger inhibitory effect than irbesartan on the expression of phosphorylated moesin, a substrate of Rho kinase (P<0.05).

CONCLUSIONAngII induces the contraction of HASMCs partially as a result of activation of Rho-kinase pathway.

Amides ; pharmacology ; Angiotensin II ; pharmacology ; Asthma ; physiopathology ; Biphenyl Compounds ; pharmacology ; Bronchi ; cytology ; Humans ; Muscle Contraction ; drug effects ; Muscle, Smooth ; cytology ; Primary Cell Culture ; Pyridines ; pharmacology ; Signal Transduction ; drug effects ; Tetrazoles ; pharmacology ; rho-Associated Kinases ; metabolism

Objective This study aimed to explore issues about dysandia female during pregnancy,peripartum and contraception and to supply reference for their reproductive health improvement. Methods Purposive sampling strategy was used. With the aid of an interview guide, 12 dysaudia women who met inclu-sion and exclusion criteria were invited to be interviewed. Interviews were recorded, validated and then ana-lyzed. Results Four themes were generated, limited knowledge, passive information acquisition, total depen-dence on family and reluctant to talk about contraception. Total dependence on family was noticeable during pregnancy and peripartum. Though little was known about health protection related to pregnancy, peripartum and contraception, participants were passive at seeking for relevant information. Conclusions Deaf women may experience more threats to reproductive health in relation to pregnancy, peripartum and contraception that they require more guidance and assistance.