Endoplasmic reticulum stress(ERS) is a kind of subcellular pathological state,and associated with many diseases.Recently,the research of ERS on chondrocyte is at the beginning stage,and may be involved in pathogenisis of rheumatoid arthritis(RA) and osteoarthritis(OA).It has been proved that ERS can interfere the differentiation of chondrocyte,decrease the synthesis of abnormal protein,attenuate the injury of cell.But overreaction of ERS can cause chondrocyte apoptosis through an independent pathway without of Fas and NO.There are three signal transmission passages in ERS:ATF-6(activating transcription factor 6)、Ire 1(inositol-requiring 1)and PERK(PKR-like ER kinase).The three protein molecules activate apoptotic genes by TRAF-2(TNF receptor-associated factor 2)and GADD153(growth arrest and DNA-damage-inducible gene 153),initiate the chondrocyte apoptosis.Therefore,ERS may effect the pathogensis of RA and OA by modulating chondrocyte function and inducing apoptosis,but more research are needed to reveal the mechanism.

Objective To investigate early changes of angiopoietin-2(Ang-2)in multiple trauma patients and assess its clinical significance.Methods Forty-five multiple trauma patients aged 2060 years admitted to the hospital within one hour after injury were randomly divided into three groups according to injury severity score(ISS).Blood specimens were obtained immediately upon arrival in the emergency department and plasma samples were assayed for comparing changes of Ang-2,TNF-a and IL-6.Meanwhile,plasma level of Ang-2 was measured and analyzed under different oxygenation index,shock index and base deficit.Results Plasma level of Ang-2 was positively correlated with ISS(P ＜ 0.01)and was concordant with the plasma levels of TNF-a and IL-6(P＜0.01).Furthermore,plasma level of Ang-2 was elevated upon increase of shock index or decrease of oxygenation index(P ＜ 0.01).Plasma level of Ang-2 was elevated with the increase of base deficit(P ＜ 0.01).Conclusions High level of Ang-2 is a marker of endothelial activation and dysfunction early after trauma.Ang-2 is related tightly with the injury severity,inflammation factors,systemic oxygenation and tissue hypoperfusion and may have a tight relation with pathophysiological development and clinical outcome after trauma.

OBJECTIVE: To investigate the effects of Wenyang Huoxue Recipe (WRHXR), a compound traditional Chinese herbal medicine for warming yang and promoting blood flow, on the expression of angiopoietin mRNA in rats with chronic aristolochic acid nephropathy induced by Caulis Aristolochia Manshuriensis (CAM) decoction, and to explore the protection mechanism of WYHXR against kidney damage. METHODS: Twenty-eight male SD rats were randomly divided into normal control group, CAM group and WYHXR-treated group. Rats in the normal control group (n=8) and CAM group (n=10) were intragastrically administered with normal saline 10 ml/(kg.d) or CAM decoction 10 ml/(kg.d) respectively. Rats in the WYHXR-treated group (n=10) were intragastrically administered with WYHXR 30 g/(kg.d) and CAM decoction 10 ml/(kg.d). The expressions of Ang-l and Ang-2 mRNAs were detected by real-time reverse transcription polymerase chain reaction (RT-PCR) after 20-week treatment. RESULTS: Compared with the normal control group, the expression of Ang-l mRNA was significantly decreased, and the expression of Ang-2 mRNA was significantly increased in the CAM group (P<0.01). Compared with the CAM group, the expression of Ang-l mRNA was increased in the WYHXR-treated group (P<0.01). The expression of Ang-2 mRNA had no significant difference between the CAM group and the WYHXR-treated group (P>0.05). Renal pathology showed that renal damage in WYHXR-treated group was significantly reduced as compared with the CAM group. CONCLUSION: WYHXR can up-regulate the expression of Ang-l mRNA, which may be its action mechanism in protecting the kidneys.

Objective:To purify and characterize the major antigenic components of sand swimming crab,and provide theoretical evidences for the research of standardization of allergenic vaccine.Methods:The crude extract of Linnaeus was prepared using classical method, and electrophoresis of SDS-PAGE was used to separate the proteins of each group. The allergen proteins of 26 crab were identified using Western blot. To distinguish between the primary allergen proteins and secondary allergen proteins, FPLC(Gel Chromatography and ion exchange chromatography) was used to filtrate and identify the allergen protein.Results:SDS-PAGE analysis revealed that sand swimming crab proteins were composed of at lease 9 discrete protein bands, which molecular weight ranging from 13 000 to 90 000. Major bands were of 20 900, 24 200, 27 100, 29 200, 33 700, 38 900, 48 700, 74 700, 89 100 respectively. Western blot assay indicated that the crude extract reacted with sera obtained from 26 crab allergenic subjects and contained 5 allergen bands altogether, and the bands of 74 400 and 48 700 were the major allergenic components. The positive rates of the two major allergen proteins were both 100%, indicating the allergenic components maintained the immunocompetence after yielding from chromatography.Conclusion:The 74 400 and 48 700 bands are the major allergens of sand swimming crab. The major allergen proteins can be purified by chromatography.

Objective To summarize the perioperative characters of femoral neck fractures in patients aged at over 80 years by analyzing the surgical treatment procedures. Methods A retrospective study was done on data of patients with femoral neck fractures treated surgically from June 2000 and June 2008. There were 25 males and 36 females, at average age of 83.4 years (80-94 years). Of all, there were 53 patients with fresh fractures including six with type Garden Ⅰ or Ⅱ fractures and 47 with type Garden Ⅲ or Ⅳ fractures, eight with old fractures ( all type Garden Ⅳ). Fifty-one patients (84%) were accompanied with preexisted internal medical or neurological diseases. Six patients with type Garden Ⅰ or Ⅱ fractures were treated with cannulated screw fixation (CSF). Among 55 patients with type Garden Ⅲ or Ⅳ fractures inclu-ding 47 fresh fractures and 8 old ones, 53 patients were treated by hip hemiarthroplasty (HHA) but the other two by cemented total hip arthroplasty (THA) because of preexisting hip osteoarthritis. Results The average operation duration and blood loss were 51 minutes and 50 ml in CSF group, 81 minutes and 180 ml in HHA group, 105 minutes and 350 ml in THA group. Fifteen patients (25%) had postoperative com-plications, mostly hypotension, cardiac insufficiency and atrial fibrillation, accounting for 12%. Postoperative hemoglobin was decreased by 11.9%, 17.1% and 18. 1% on average respectively in CSF group, HHA group and THA group, and postoperative albumin decreased by 10.8%, 18. 1% and 20.2% on average respectively in CSF group, HHA group and THA group. There were 17 patients (28%) with hemoglobin below 100 g/L and four (7%) with albumin below 30 g/L after operation. Conclusions The patients aged ≥80 years with femoral neck fractures are usually accompanied with preexisted internal medical or neurological diseases and have high risk in operation. Adequate preoperative preparation and active prevention and treatment of perioperative complications are important for successful operation and good postoperative results.

Objective To identify hepatitis C virus(HCV)-specific cytotoxicity T lymphocytes (CTL)epitopes by the combination of T epitopes prediction software and in vitro assays.Methods HCVspecific CTL epitopes were predicted by T epitope prediction software Rankpep and then candidate HCV-specific CTL epitopes were selected.Candidate HCV-specific CTL epitopes were used to stimulate PBMC of HCV-infected patients and healthy volunteers.and then enzyme-linked immunospot(ELISPOT)and intracellular cytokine staining(ICS)were used to measure the frequencies of IFN-γ-producing cells in total PBMC and the percentages of IFN-γ+CD8+T cells in total CD8+T cells,respectively.Results Five candidate CTL epitopes[NS3 450(TVPQDAVSR),NS3 594(GPTLLYRL),Ns4b 78(sMMAFSAAL),NS5a 416(SEENVSVVF)and NS5a 367(TVSSALAEL)]were used to stimulate PBMC of ten HCV-infected patients and two healthy volunteers.PBMC of seven HCV-infected patients secreted IFN-γ while PBMC of healthy volunteers did not produce IFN-γ.The frequencies of peptide-specific IFN-γ-producing cells ranged from 5 to 36 SFC/105 PBMC and the percentages of peptide-specific IFN-γ+CD8+T cells ranged from 0.02%-0.25%.Conclusion Resuhs of ELISPOT assay and ICS assay confirm that these five peptides NS3 450,NS3 594,NS4b 78,NS5a 416 and NS5a 367 are identified as Hovel HCV-specific CTL epitopes.

Objective To identify human leucocyte antigen (HLA)-A* 0201-restricted hepatitis C virus (HCV)-cytotoxic T lymphocyte (CTL) epitopes. Methods Based on the prediction results of RANKpep and SYFPEITHI prediction programs, six candidate CTL epitopes were selected and synthesized. The affinity of candidate CTL epitopes to HLA-A* 0201 molecules of T2 cells was explored. Subsequently, enzyme-linked immunosorbent spot (ELISPOT) assay and intracellular cytokine staining (ICS) were utilized to determine whether candidate CTL epitopes could induce the recall positive response in peripheral blood mononuclear cells (PBMC) of HLA-A* 0201 positive HCV-1b-infected patients. Results Among six candidate CTL epitopes, peptides C_181(LLSCLTTPV) and NS2_172 (VLQAGLIRV) had high affinity to HLA-A* 0201 molecules. Moreover, the affinity was proportional to the concentration of peptide. Furthermore, among ten HLA-A* 0201 positive HCV-1b-infected patients, the frequencies of C_181 and NS2_172-specific interferon (IFN)-γ-producing cells were 0-19 spots forming cells (SFC)/1 × 105 PBMC and 0-20 SFC/1 × 105 PBMC, respectively.The percentages of C_ 181 and NS2_172-specific IFN-γ+ CD8+ T lymphocytes in total CD8+ T lymphocytes were 0.006%-0.065% and 0.005%-0.080%, respectively. Conclusion Peptides C_181 (LLSCLTTPV) and NS2_172 (VLQAGLIRV) are identified as novel HLA-A* 0201-restricted HCV-CTL epitopes.

A label-free electrochemical immunosensor using hollow structure nanomaterials based on its ordered porous and big surface area was designed. Au nanocage, with good conductivity, catalysis, and biocompatibility, was prepared and modified on the surface of glassy carbon electrode with graphene to immobilize antibody of microcystin directly. In the absence of microcystin, biosensor can obtain high current response signal of electrochemical probe ( [ Fe( CN) 6 ] 3-/4-. When microcystin was combined with its antibody specifically, the charge density and mass transfer resistance on the surface of electrode increased, resulting in a decrease of the corresponding peak current of [ Fe ( CN ) 6 ] 3-/4-. This change was in proportion to the concentration of microcystin indirectly. Experiment conditions such as cultivation time of antigen and concentration of antibody were optimized. The results showed wide linear range of 0. 05 μg/L-1. 0 mg/L and the detection limit of 0. 017 ng/mL. This sensor has good stability and simple production procedure. This sensor provides a new and simple means for the ultrasensitive determination of microcystins in real water samples.

Objective To investigate the dynamic change and the clinical curative effect evaluation of plasma (1-3)-beta glucan D (BG) in the patients with pulmonary disease complicating fungal infection .Methods The MB-80 miroorganism dynamic rapid de-tection system and fungi BG detection kits were adopted to detect plasma BG content before and after treatment in 87 cases of pul-monary disease complicating fungal infection and the controls .The sputum culture in the patients was performed before and after treatment .Results Plasma BG levels before antifungal therapy ,at 1 ,2 weeks after treatment in 87 patients were (162 .81 ± 70 .03) , (15 .89 ± 30 .88) and (4 .58 ± 7 .87)pg/mL ,which in the control group was (5 .62 ± 1 .83)pg/mL ,plasma BG level had statistical differences between before treatment and at 1 ,2 weeks after treatment in the patients with the control group (P<0 .05);Plasma BG levels between at 1 week after treatment with at 2 weeks after treatment and the control group had statistically significant differ-ences (P<0 .05) .Among 87 patients ,66 cases were positive sputum culture at 1 week after antifungal drug treatment and 9 cases were positive sputum culture at 2 weeks after treatment .Conclusion Continuously monitoring the patient′s plasma BG level com-bined with the sputum fungal culture results ,clinical symptoms and lung shadow in X-ray has certain clinical value to judge the anti-fungal effect .

Objective To explore the feasibility of establishment of NOD/SCID-mouse model with multiple myeloma by using plasma cells from myeloma patients.Methods The femurs and tibias were removed from the New Zealand white rabbits;the muscles,periosteum and cartilage tissues were cleared.Then each bone was cut into two pieces gently along its middle.The NOD/SCID mice weighing 25 - 30 g (4 - 6 weeks)were anesthetized by intraperitoneal injection;rabbit bone was inserted into the right side of the mouse back and engraftment of the bones was allowed to take place after 4 weeks.The 5000 000 purified plasma cells which expressed CD38 +/CD45 - were immunofluorescence labeled and then injected slowly into the implanted rabbit bone through the distal end.The mice were observed weekly;the plasma cells growth in mice was screened by the living-imaging system and the tumor from the mice was determined by biopsy.Results The implanted rabbit bone survived after 4 weeks.The tumor in mice was observed 2 weeks after the purified myeloma cells were injected into the rabbit bone,and it reached 100 mm3 after 8 weeks.Results of the living-imaging system showed that the myeloma cells had uptake in the rabbit bone after 2 weeks of injection and this phenomenon was more pronounced after 8 weeks of injection (2.4×10 4 vs .1.5× 10 5 ,P < 0.05 ).The tumor infiltrated with numerous plasma cells and osteoclasts increased upon the biopsy. Conclusion Rabbit bone marrow implanted into NOD/SCID mice can effectively support local injection of plasma cells of multiple myeloma patients,and the NOD/SCID-mouse model of myeloma has been established.This model can be used to study in vivo experiments related to myeloma and clinical therapeutic approaches for this disease.