BACKGROUNDCalreticulin (CRT) is major Ca 2+ -binding chaperone mainly resident in the endoplasmic reticulum (ER) lumen. Recently, it has been shown that non-ER CRT regulates a wide array of cellular responses. We previously found that CRT was up-regulated during hypoxia/reoxygenation (H/R) and this study was aimed to investigate whether CRT nuclear translocation aggravates ER stress (ERS)-associated apoptosis during H/R injury in neonatal rat cardiomyocytes.

METHODSApoptosis rate and lactate dehydrogenase (LDH) leakage in culture medium were measured as indices of cell injury. Immunofluorescence staining showed the morphological changes of ER and intracellular translocation of CRT. Western blotting or reverse transcription polymerase chain reaction was used to detect the expression of target molecules.

RESULTSCompared with control, H/R increased apoptosis rate and LDH activity. The ER became condensed and bubbled, and CRT translocated to the nucleus. Western blotting showed up-regulation of CRT, Nrf2, activating transcription factor 4 (ATF4), CHOP and caspase-12 expression after H/R. Exogenous CRT overexpression induced by plasmid transfection before H/R increased cell apoptosis, LDH leakage, ER disorder, CRT nuclear translocation and the expression of ERS-associated molecules. However, administration of the ERS inhibitor, taurine, or CRT siRNA alleviated cell injury, ER disorder, and inhibited ERS-associated apoptosis.

CONCLUSIONSOur results indicated that during H/R stress, CRT translocation increases cell apoptosis and LDH leakage, aggravates ER disorder, up-regulates expression of nuclear transcription factors, Nrf2 and ATF4, and activates ERS-associated apoptosis.

Animals ; Apoptosis ; genetics ; physiology ; Calreticulin ; genetics ; metabolism ; Cell Hypoxia ; genetics ; physiology ; Cell Survival ; genetics ; physiology ; Cells, Cultured ; Endoplasmic Reticulum Stress ; physiology ; Myocytes, Cardiac ; cytology ; metabolism ; RNA Interference ; Rats

Chemotherapy, Adjuvant ; Humans ; Stomach Neoplasms ; drug therapy

Objective To purify rabbit anti-hRBBP10 polyclonal antibody.Methods The recombinant fusion protein PTC-hRBBP10 was expressed in the E.coli and was purified through amylose resin chromatography column and superose 12 gel fitration .The purified PTC-hRBBP10 was coupled to the NHS-activated sepharoseTM to prepare affinity chromatography column to purify rabbit anti-hRBBP10 polyclonal antibody. Results ① PTC-hRBBP10 was expressed and purified successfully with relative molecular mass(Mr) of 80?103 and its purity could reach about 95%.② Purified Rabbit anti-hRBBP10 polyclonal antibody could bind Specifically to PTC-hRBBP10.Conclusions With better specificity, The purified rabbit anti-hRBBP10 polyclonal antibody provide condition for studying the function of the protein, and helps to develop new techniques of tumor diagnosis and treatment.

0.05),but the proportion of NRDS,the rate of mechanical ventilation dependence,and mortality had significant diffe-rences(Pa

ObjectiveTo study the occurrence and influence factors of drug-induced injuries of vital organs caused by traditional Chinese medicine injection.MethodsChina Hospital Knowledge Database (CHKD) from 1993-2013 and case reports concerning adverse effects and drug-induced diseases caused by traditional Chinese medicine injection from Wanfang Data Knowledge Service Platformwere retrieved. Indicator systemwasestablished and relevant contentsweresummarized and analyzed.Results34 types of traditional Chinese medicine injectionswereincluded and 699 drug-induced diseasesweresummarized, taking up 53.98% of the total adverse effects. Among them, the top three included in the drug-induced diseaseswere acanthopanax injection, safflower injection and Mailuoning injection. The non-conformance between the traditional Chinese medicine injections in the literature and instructions mainly reflects unreasonable solvent selection, large compatibility concentration and usage and dosage beyond those specified in the instructions. ConclusionThe occurrence of drug-induced injuries of vital organs caused by traditional Chinese medicine injectionis relatively high, drug instructions shall be strictly followed in accordance with requirements during clinical application.

Objective To label human VEGF targeted bevacizumab (avastin) with 111In and to evaluate the application of 111 In?DTPA?avastin SPECT imaging for tumor diagnosis. Methods DTPA?avastin was prepared by coupling with a bifunctional chelating agent, and then labeled with 111 In to obtain 111 In?DTPA?avastin. The stability and molecular integrity of the labeled radiotracer were studied. Human hepatoma cell ( BEL7404) bearing nude mice tumor model was employed for tumor targeting evaluation. Gamma imaging was acquired after intravenous injection of 18.5 MBq probe. At the end of the observation, animals were sac?rificed for bio?distribution study. Results 111 In?DTPA?avastin tracer was synthesized and purified to a?chieve a radiochemical purity yield above 98% and specific activity up to 185 GBq/nmol. Its stability in 5%BSA was optimal, and the radiochemical purity after incubation for 96 h was over 90%. Gamma imaging re?sults showed that the tracer possessed definite tumor targeting property. Its biodistribution was consistent with that of normal in vivo antibody metabolism while possessing a good tumor?targeting property with a relatively high uptake of (3.8±0.8) %ID/g in tumor tissues 96 h after injection. Conclusions 111 In?DTPA?avastin tracer has good physicochemical properties, in vivo stability and good VEGF targeted binding. 111 In?DTPA?avastin has potential to be a new molecular probe for SPECT imaging.

Objective To explore the significance of detecting IgH gene rearrangement in diagnosis of non-Hodgkin lymphoma. Methods By using family-specific primers mixtures (VH1,VH2,VH3) for IgH FR3A,FR2A,FR1 genes,clonal IgH gene rearrangement was detected by heminested and multiplex PCR in 44 cases of B-NHL,1 undifferentiated lymphoma,15 T-NHL and 5 lymphonode reactive hyperplasia. Results Clonal IgH gene rearrangement was detected in 36 of 44 B-NHL (positive rate 82%) by using FR3A primers,27 of 44 B-NHL (positive rate 61%) by using FR2A primers. By using FR1 family-specific primers mixtures (VH1,VH2,VH3) and JH (JHb,JHc),clonal IgH gene rearrangement was demonstated respectively in 34/33 of 44 B-NHL (positive rate 77%,75%). The total positive rate was 95% (42/44) by combined family-specific primers mixtures (VH1,VH2,VH3) of FR3A,FR2A,FR1 gene. Four of 15 T-NHL revealed clonal IgH gene rearrangement,while 5 LRH had no clonal IgH gene rearrangement. One NHL unclassified by immunohistochemical methods was classified definitely as B cell-derived lymphoma by PCR. Conclusion The detection rate of IgH gene rearrangement can be improved by combined usage of several pairs of primers,especially family-specific primers. Detecting clonal IgH gene rearrangement is significant in the procedure of differential diagnosis for B/T-NHL and reactive hyperplasia.

0.05). Conclusion Enough high-purified LECs can be isolated by collagenase digestion procedure followed by immunomagnetic beads sorting. Post-thawed endothelial cells are proved to have high vitality and growth potential in vitro without significant morphological changes. Cryopreserved LECs may serve as a cell choice for research of lymphangiogenesis and lymphatic patterning.

OBJECTIVETo explore the effective method of the prevention and treatment of procedural pain in dressing changes of burn wounds.

METHODSNinety patients of burn injury were randomized into 3 groups, 30 cases in each one. In the group A, fentanyl citrate injection was used at corresponding injury area, jiaogan (AH6a, sympathetic nerve), fei (CO14, lung), neifenmi (CO18, endocrine) on ear, 0.25 mL at each point. In the group B, fentanyl citrate injection was applied subcutaneously in the deltoid muscle, 1 mL. In the group C, 0.9% sodium chloride injection was applied subcutaneously in the deltoid muscle, 1 mL. The visual analogue scale (VAS) was used to evaluate the analgesic effect before, during and 10 min after dressing change in the patients of the three groups separately.

RESULTSIt was not different in VAS score before dressing change among the three groups (P> 0.05). Compared with that before dressing change, the pain was not significant and VAS score was not different during and after dressing change in the patients of the group A (both P>0.05), but the score in the patients of the group B and C was different significantly (all P<0.05). The VAS score during and after dressing change in the group A was lower than that in the group B and C (all P<0.05), and the score in the group B was lower than that in the group C (P<0.05).

CONCLUSIONFentanyl injection of small dose at auricular points achieves definite analgesic effect on procedural pain in dressing changes of burn wounds, superior to subcutaneous injection of fentanyl.

Acupuncture Points ; Adolescent ; Adult ; Aged ; Burns ; complications ; therapy ; Female ; Fentanyl ; administration & dosage ; Humans ; Male ; Middle Aged ; Pain ; drug therapy ; etiology ; Pain Measurement ; Young Adult

AIM To prepare diosgenin nanosuspensions.METHODS The nanosuspensions prepared by media milling method were solidificated by freeze drying method.With particle size and polydispersity index (PDI) as evaluation indices,stabilizer kind,ratio of diosgenin to stabilizer,ratio of preliminary nanosuspension volume to grinding bead amount,milling time,lyoprotectant kind and its amount as influencing factors,single factor test was applied to screening preparation and solidification processes.The morphology of nanosuspensions was observed,then the particle sizes and polydispersity indices of nanosuspensions and freeze-dried powder were determined.RESULTS The optimal conditions were determined to be 6:1 for ratio of diosgenin to Pluronic F127 (stabilizer Ⅰ),50:1 for ratio of diogenin to sodium dodecyl sulfate (SDS,stabilizer Ⅱ),1:4 for ratio of preliminary nanosuspension volume to grinding bead amount,120 min for milling time,and 8％ PEG-6000 and 2％ mannitol as lyoprotectants.The average particle size and polydispersity index of rod-like or flaky nanosuspensions were (348.1 ±14.2) nm and 0.244 ± 0.059,respectively,which were lower than those of freeze-dried powder.At room temperature,the particle sizes of nanosuspensions and freeze-dried powder remained stable within 35 d and 3 months,respectively.CONCLUSION The physical stability of diosgenin freeze-dried powder is superior to that of its nanosuspensions,which can be used after being reconstituted.