BACKGROUNDTo express in vitro the bovine enterokinase catalytic subunit (EKL) protein, which could be used in the future for the cleavage and purification of fusion proteins.

METHODSBovine enterokinase catalytic subunit cDNA was obtained by RT-PCR from duodenal mucosa of a bovine obtained at wholesale market, and then cloned into a pUCmT cloning vector and sequenced. The desired gene fragment was inserted into a pET39b expression plasmid and the recombinant vector pET39b-EKL was transformed into E. coli BL21 (DE3). Protein expression was induced using IPTG. The recombinant DsbA-EKL was purified with His.Tag affinity chromatography, and it bioactivity was analyzed.

RESULTSCompared with the sequence deposited in GenBank, the sequence of the EKL gene cloned in the present study is correct. It was also confirmed that the nucleotide sequence of expression plasmid pET39b-EKL was correct at the conjunction site between the recombinant DNA 5' terminal multi-cloning site and the recombinant fragment. SDS-PAGE analysis indicated that the target product was about 65 kDa and represented 28% of total cell protein. Purified recombinant protein was obtained by metal chelating chromatography using Ni-IDA resin. After desalting and changing the buffer, the crude kinase was incubated at 21 degrees C overnight and shown to have a high autocatalytic cleavage activity.

CONCLUSIONSThe EKL gene from Chinese bovine has been cloned successfully and expressed. This investigation has layed the foundation for future enterokinase activity research and for further large-scale application of expression products.

Animals ; Catalytic Domain ; genetics ; Cattle ; Cloning, Organism ; DNA, Complementary ; Enteropeptidase ; analysis ; genetics ; Recombinant Proteins

OBJECTIVE Zn-doped CuO nanocomposites (nZn-CuO NPs) are novel nanoparticles synthesized by sonochemical method.This study aimed to further investigate the antitumor effects and mechanism of nZn-CuO NPs, as well as the exact mechanism of reactive oxygen species (ROS) on nZn-CuO NPs-induced death using N-acetylcysteine (NAC). METHODS The antitumor effects of nZn-CuO NPs were evaluated by MTS assay and orthotopic transplantation tumor model in nude mice. The effects of nZn-CuO NPs with or without NAC on ROS production, DNA damage, apoptosis, mitochondrial damage, autophagy, lysosome impairment, and ER and Golgi stress were determined. Also,western blot was used to detect apoptosis and autophagy related proteins,as well as NF-κB pathway related proteins. RESULTS nZn-CuO NPs significantly inhibit tumor growth both in vitro and in vivo. nZn-CuO NPs were able to cause cytotoxicity, ROS production, DAN damage mitochondrial damage, apoptosis, and autophagy, and NAC can attenuate them. Further studies showed that nZn-CuO NPs induced changes of apoptosis, autophagy and NF-κB pathway related proteins, and NAC can restore them. CONCLUSION Overall, our data demonstrated that nZn-CuO NPs could inhibit tumor growth both in vitro and in vivo by ROS-dependent regulation of apoptosis and autophagy, which might be cross-linked by NF-κB pathways.

Objective To explore the change of nerve growth factors(NGF) through the blood brain barrier(BBB) after distal intravenous injection of mannitol into the experimental rats and the effect of exogenous NGF on the expression of growth associated protein-43 in hypoxic-ischemic brain.Methods One hundred cases of 7 days rats were divided into 2 units.One unit was divided into 3 groups:treatment group,control group and sham operated group,20 rats in each group.The other unit was divided into 4 groups:mannitol and NGF treated group,NGF treated group,control group,and sham operated group,there were 10 rats in each group.The model rats with perinatal hypoxic-ischemic brain damage(HIBD) rats were prepared by ligation of left common carotid artery with a temporary systemic hypoxia(inhaling 80 mL/L O2 and 920 mL/L N2).The sections of brains were processed by immunochemistry with antibodies against GAP-43,and the study and memory ability of rats were tested by maze test.The effect of osmotic opening of BBB on the facilitation of NGF′s passage was tested by ELISA.Results The expression of GAP-43 increased after NGF treatment,and the differences were remarkable(P

Objective To investigate the effect of early rehabilitation on neurological function and cerebral blood flow of patients with acute cerebral infarction.Methods100 cases of acute cerebral infarction were randomly divided into the rehabilitation group(53 cases) and control group(47 cases).Both groups received the routine treatment;besides,patients in the rehabilitation group were treated with early rehabilitation training.The treatment efficacy was assessed by neurological function deficit evaluation,and the cerebral blood flow was measured by transcranial Doppler(TCD) before and 30 days after treatment.ResultsThe scores of neurological function deficit evaluation of all patients in each group decreased after treatment,but the scores of the rehabilitation group were obviously lower than that of the control group(P<0.01);and the results of TCD of the rehabilitation group were also superior to that of the control group(P<0.01).ConclusionEarly rehabilitation training may obviously improve neurological function and cerebral blood flow of patients with acute cerebral infarction.

Sixteen compounds were isolated from the ethanol extract of Illigera rhodantha by silica gel, ODS, and Sephadex LH-20 column chromatographies. These compounds were identified as (2R,3R)-2,3-dihydroxy-2-methylbutane-1,4-diyldibenzoate (1), p-hydroxyphenethyl trans-ferulate (2), 4-O-benzoyl-2-C-methyl-D-erythritol (3), N-trans feruloyl-3-methyldopamine (4), tribulusamide A (5), cryptomeridiol (6), teuclatriol (7), oleanolic acid (8), icario A₂ (9), vanillic acid (10), p-hydroxybenzoic acid (11), gallic acid (12), ethyl gallate (13), chrysophanol (14), D-1-O-methyl-inositol (15), and hexadecanoic acid (16) based on their spectral data and physico-chemical properties. Compound 1 is an undescribed compound, of which its absolute configurations were determined by Mosher and ROESY methods; all the compounds except 10, 11 and 14 were isolated from Illigera genus for the first time. Compared with the positive control indomethacin, compounds 4-6, 8 and 9 inhibited significantly against the NO production in LPS-induced RAW 264.7 cells.

AIMTo detect the residual clenbuterol in pork meat and liver using HPLC with Coulometric electrode array system.

METHODSHomogenized meat or liver sample was treated with 1 mol x L(-1) hydrochloric acid and centrifuged, the fat existing in meat or liver tissue was removed by diethyl ether. The pH of the remaining aqueous layer was adjusted to 10.8 +/- 0.2 or 11.6 +/- 0.2 for meat or liver and liquid-liquid extraction with diethyl ether was followed. The ether extract was evaporated to dryness, the residue was dissolved in the mobile phase. The mobile phase A consisted of 50 mmol x L(-1) phosphoric acid-30 mmol x L(-1) triethylamine and was adjusted to pH 4.0 with 2 mol x L(-1) sodium hydroxide solution. The mobile phase B consisted of methanol-acetonitrile (30:45). A mixture of mobile phase A and B (80:20) was used in the method. A four electrode array module was selected for quantitation, the electrode potentials were set at 450, 600, 650 and 680 mV respectively.

RESULTSThe two calibration curves for meat and liver showed good linearity between 1.88 - 60.16 ng x g(-1), the detection limit of clenbuterol was 1.2 ng x g(-1).

CONCLUSIONThis method using HPLC-electrochemical detection is reproducible, and the sensitivity is good enough for the determination of clenbuterol in meat and liver.

Animal Feed ; analysis ; Animals ; Chromatography, High Pressure Liquid ; methods ; Clenbuterol ; analysis ; Drug Residues ; analysis ; Electrochemistry ; methods ; Electrodes ; Liver ; chemistry ; Meat ; analysis ; Swine

OBJECTIVETo study the pathologic features, diagnosis and differential diagnosis of interdigitating dendritic cell sarcoma (IDCS).

METHODSThe clinical findings, morphologic features and immunophenotype of 3 cases of IDCS were investigated.

RESULTSGross examination showed that IDCS had a greyish-white to greyish-yellow cut surface. The site of occurrence included lung, spleen (with lymph node metastasis) and lymph node. Histologically, the tumor cells were arranged in nests, fascicles and whorls, with intimate admixture of many lymphocytes and plasma cells. They were oval to spindle in shape and contained pale eosinophilic cytoplasm, oval and sometimes grooved nuclei, small distinct nucleoli and ill-defined cell borders. Immunohistochemical study showed that the tumor cells expressed S-100 protein.

CONCLUSIONSIDCS is a rare type of histiocytic and dendritic cell malignancy with distinctive morphologic findings. It needs to be distinguished from follicular dendritic cell sarcoma, inflammatory pseudotumor, Langerhans' cell histiocytosis, malignant melanoma, undifferentiated carcinoma and anaplastic large cell lymphoma. Immunohistochemical staining for S-100 protein is helpful in confirming the diagnosis.

Adolescent ; Carcinoma ; pathology ; Dendritic Cell Sarcoma, Follicular ; pathology ; Dendritic Cell Sarcoma, Interdigitating ; diagnosis ; pathology ; Dendritic Cells ; pathology ; Diagnosis, Differential ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; S100 Proteins ; analysis ; immunology ; Young Adult

OBJECTIVETo observe the outcome after sacral canal injection in patients with disc herniation associated with without sciatica.

METHODSFrom December 2010 to June 2011, 65 patients with acute low back pain without sciatica due to lumbar disc herniation or bulging confirmed by CT or MRI were randomly divided into sacral canal injection group (experimental group) and lumbar oblique wrench group (control group): the experimental group had 35 cases, including 30 males and 5 females, with an average age of (43.90 +/- 1.14) years old ranging from 33 to 56 years old. The control group had 30 cases, including 27 males and 3 females,with an average age of (44.00 +/- 1.19) years old ranging from 34 to 57 years old. The course of morbidity was 1 to 3 days. All patients received sacral canal injection or lumbar oblique wrench method. The visual analog scale (VAS) scores before and at 30 min after treatment were compared between two groups.

RESULTSThe symptom of acute low back pain were relieved obviously. The average VAS scores before and after treatment in experimental group were decreased from 6.63 +/- 0.97 to 3.06 +/- 1.51,in control group were from 6.67 +/- 0.96 to 3.93 +/- 1.20 respectively. These two methods could improve the VAS score,but the effect of sacral canal injection group was better than that of lumbar oblique wrench group, there was statistically differences (P < 0.05).

CONCLUSIONIt is effective that the methods of sacral canal injection and lumbar oblique wrench applied to patients with acute low back pain without sciatica due to lumbar disc herniation or bulging confirmed, the former has better effect.

Adult ; Female ; Humans ; Injections, Spinal ; Intervertebral Disc Displacement ; drug therapy ; Lidocaine ; administration & dosage ; Lumbar Vertebrae ; Male ; Middle Aged ; Prednisone ; administration & dosage ; Sacrococcygeal Region ; Visual Analog Scale

OBJECTIVETo investigate the diagnosis and differential diagnosis of extranodal Rosai-Dorfman disease.

METHODSTwo cases of extranodal Rosai-Dorfman disease were studied using hematoxylin-eosin, and immunohistochemical staining, along with a literature review.

RESULTSThe lesions of RDD were characterized by the presence of large histiocytes with emperipolesis, accompanied by infiltration of lymphocytes, plasma cells and other inflammatory cells. The large histiocytes had an abundant cytoplasm, pale to eosinophilic in appearance, positive for S-100 protein staining, with a vesicular nucleus and a small basophilic nucleolus in each cell.

CONCLUSIONSExtranodal Rosai-Dorfman disease is known as an idiopathic proliferative disease of histiocytes with a distinct morphologic feature and is very rare. Differential diagnosis from other types of fibrohistiocytic proliferation lesions is recommended.

Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Brain ; pathology ; surgery ; Brain Diseases ; metabolism ; pathology ; surgery ; Dermatologic Surgical Procedures ; Diagnosis, Differential ; Histiocytosis, Sinus ; metabolism ; pathology ; surgery ; Humans ; Male ; Middle Aged ; S100 Proteins ; metabolism ; Skin ; pathology ; Skin Diseases ; metabolism ; pathology ; surgery

BACKGROUND: This meta-analysis aimed to determine whether extracorporeal cardiopulmonary resuscitation (ECPR), compared with conventional cardiopulmonary resuscitation (CCPR), improves outcomes in adult patients with cardiac arrest (CA). DATA RESOURCES: PubMed, EMBASE, Web of Science, and China Biological Medicine Database were searched for relevant articles. The baseline information and outcome data (survival, good neurological outcome at discharge, at 3–6 months, and at 1 year after CA) were collected and extracted by two authors. Pooled risk ratios (RRs) and 95% confidence intervals (CIs) were calculated using Review Manager 5.3. RESULTS: In six studies 2260 patients were enrol ed to study the survival rate to discharge and long-term neurological outcome published since 2000. A significant effect of ECPR was observed on survival rate to discharge compared to CCPR in CA patients (RR 2.37, 95%CI 1.63–3.45, P<0.001), and patients who underwent ECPR had a better long-term neurological outcome than those who received CCPR (RR 2.79, 95%CI 1.96–3.97, P<0.001). In subgroup analysis, there was a significant difference in survival to discharge favoring ECPR over CCPR group in OHCA patients (RR 2.69, 95%CI 1.48–4.91, P=0.001). However, no significant difference was found in IHCA patients (RR 1.84, 95%CI 0.91–3.73, P=0.09). CONCLUSION: ECPR showed a beneficial effect on survival rate to discharge and long-term neurological outcome over CCPR in adult patients with CA.