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 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.

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.

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.

Objective To study the relationship between myofibril fragmentation index (MFI) of human skeletal muscle and postmortem interval (PMI). Methods The protein concentrations of human right bi-ceps brachii muscle and right quadriceps femoris muscle were obtained at different PMI, and detected at room temperature by biuret method. The MFI of skeletal muscle at 540 nm was measured by ultraviolet spectrophotometer. Regression analysis was performed with time of death as independent variable (x) and MFI as dependent variable (y). Results In early PMI, the MFI of human skeletal muscle increased obviously according to the prolongation of PMI, and peaking by 12 h and then tended to steady. Within 12 h after death, the regression equations of right biceps brachii muscle and right quadriceps femoris muscle were y=32.660+3.227 x(r=0.9879) and y=32.380+3.495 x(r=0.9839), respectively. Conclusion There's high correlation between MFI and PMI. Combining with forensic practice, MFI can be used for the estimation of early PMI (especially in 12 h).

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.

Bullous pemphigoid (BP) is the most common autoimmune subepidermal blistering skin disease,and a large number of eosinophils (EOS) obviously infiltrate the dermis of BP lesions.Additionally,EOS and their activated cytokines and chemokines are abundantly present in blisters and peripheral blood of BP patients.It is also proved that EOS can induce the dermal-epidermal separation.The level of interleukin (IL)-5 is markedly increased in the sera and blister fluids of BP patients,and IL-5 secreted by Th2 cells and EOS can regulate the differentiation,activation and survival of EOS.All the above evidence indicates that EOS are associated with the occurrence of BP.Researches on EOS and their cytokines will develop a new field of targeted therapy for BP.

OBJECTIVETo investigate the expression characteristics of human beta-defensin-2 (HBD-2) mRNA and protein in salivary gland benign and malignant tumor tissues, as well as in salivary gland inflammation.

METHODSThe expression of HBD-2 in salivary gland benign tumor, salivary gland cancer, inflammation tissues and normal salivary gland tissues were detected by polymerase chain reaction (PCR), real-time polymerase chain reaction (Real-Time PCR) and immunohistochemical. The differences expression of HBD-2 mRNA and protein were analyzed.

RESULTSRT-PCR results showed that HBD-2 mRNA expression in salivary gland benign tumors, salivary gland cancer, and inflammation tissues was 6.468-, 0.334-, and 10.563-fold higher than that in normal tissues, respectively (P < 0.05). HBD-2 was expressed in the nuclei of these organs and malignant tissues.

CONCLUSIONHBD-2 mRNA and protein expressions are significantly increased in salivary gland benign tumor tissues and inflammation tissues compared with those in normal salivary gland tissues, but are significantly decreased in salivary gland cancer. The protein nuclear transfer in salivary gland cancer tissues is also significantly increased.

Humans ; Inflammation ; Polymerase Chain Reaction ; RNA, Messenger ; Salivary Gland Neoplasms ; Salivary Glands ; beta-Defensins ; biosynthesis