Hirudin (HV) is known as the most potent and specific inhibitor of thrombin. Although hirudin has many advantages , it has the bleeding side effect and this is the great shortage of hiudin for clinical application. In order to alleviate bleeding side effect of hirudin, fusion protein, named as FHV (fusion hirudin linked with FXa recognition peptide) was designed. The fusion protein gene ( fhv) was cloned into plasmid pPIC9K. FHV engineered Pichia pastoris containing high copies was chosen for fermentation and purification at 30 L fermentor scale, finally, FHV with purity of above 97% was obtained. To investigate the function of FHV in vivo, mouse tail thrombosis model was used. In the mice thrombus tail model induced by carrageenan, FHV decreased the length of tail thrombus significantly, similar to that of HV control, and had no obvious effects on the TT, PT and APTT. In conclusion, FHV is constructed and expressed in yeast. FHV fusion proteins is obtained by fermentation and purification. FHV has antithrombotic effects not influencing IT, PT and APTT after administration immediately in animal models. Therefore, FHV is a promising anticoagulant and antithrombotic drug.

BACKGROUNDMajority of the research on cardiac arrest (CA) have focused on post-CA brain injury and myocardial dysfunction, the renal dysfunction and acute kidney injury (AKI) in other critical illnesses after CA have not been well described. This study was designed to assess AKI with renal Doppler and novel AKI biomarkers in a swine model of ventricular fibrillation cardiac arrest (VFCA).

METHODSThirty healthy piglets were divided into VFCA group (n = 22) and Sham group (n = 8) in a blinded manner. Mean arterial pressure, heart rate, and cardiac output were recorded continuously. Cardiac arrest (CA) was induced by programmed electric stimulation in the VFCA group, and then cardiopulmonary resuscitation was performed. Twenty piglets returned of spontaneous circulation (ROSC) and received intensive care. Blood and urine samples were collected for AKI biomarkers testing, and Color Doppler flow imaging was performed at baseline, 6 h, 12 h, and 24 h, respectively after ROSC. At ROSC 24 h, the animals were sacrificed and a semi-quantitative evaluation of pathologic kidney injury was performed.

RESULTSIn the VFCA group, corrected resistive index (cRI) increased from 0.47 ± 0.03 to 0.64 ± 0.06, and pulsatility index (PI) decreased from 0.82 ± 0.03 to 0.68 ± 0.04 after ROSC. Cystatin C (CysC) in both serum and urine samples increased at ROSC 6 h, but neutrophil gelatinase-associated lipocalin (NGAL) in serum increased to 5.34 ± 1.68 ng/ml at ROSC 6 h, and then decreased to 3.16 ± 0.69 ng/ml at ROSC 24 h while CysC increasing constantly. According to the renal histopathology, 18 of 20 animals suffered from kidney injury. The grade of renal injury was highly correlated with RI, cRI, NGAL, and CysC. Linear regression equation was established: Grade of renal injury = 0.002 × serum CysC + 6.489 × PI + 4.544 × cRI - 8.358 (r2 = 0.698, F = 18.506, P < 0.001).

CONCLUSIONSAKI is common in post-CA syndrome. Renal Doppler and novel AKI biomarkers in serum and urine are of significant importance as early predictors of post-CA AKI.

Acute Kidney Injury ; blood ; etiology ; Animals ; Biomarkers ; blood ; Cystatin C ; blood ; Disease Models, Animal ; Female ; Heart Arrest ; blood ; complications ; Lipocalins ; blood ; Male ; Swine ; Ultrasonography, Doppler ; methods ; Ventricular Fibrillation ; blood ; complications

OBJECTIVETo study the protective effect of Shenxiong injection on the cerebral ischemia-reperfusion injury of senile rats.

METHODTotally 108 Sprague-Dawley (SD) rats were randomly divided into the sham operation group, the model group, the Ni-modipine group and Shenxiong injection groups (low, middle, and high doses). The rat brain ischemia-reperfusion model was established by the middle cerebral artery occlusion (MCAO) method in rats, in order to observe the effect of Shenxiong injection on neurological score and brain infarct volume of rats with cerebral ischemia-reperfusion injury, and determine the contents of NOS, NO, SOD, MDA and LDH in brain tissues. The contents of TNF-alpha and IL-1beta levels in brain tissues were measured by enzyme-linked immunosorbent assay (ELISA) method.

RESULTShenxiong injection could significantly decrease neurological score, injury degree of brain tissues and brain infarct volume of rats with cerebral ischemia-reperfusion injury, increase the vigor of SOD, decrease the levels of MDA, NO, NOS and LDH, and inhibit IL-1beta and TNF-alpha expressions.

CONCLUSIONShenxiong injection has the obvious protective effect on the brain ischemia-reperfusion injury in rats. Its mechanism may be related to the improvement of neurological function, the reduction of free radical injury, and the inhibition of inflammation factor expression.

Animals ; Brain ; blood supply ; drug effects ; metabolism ; Brain Ischemia ; complications ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Injections ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; complications ; drug therapy ; enzymology ; metabolism ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the protective effect of Astragalus Membranaceus Injection on human umbilical vein endothelial cells (HUVECs) against tumor necrosis factor alpha (TNF-alpha).

METHODSCultured passage 2 HUVECs were stimulated with TNF-alpha with or without a 2-h Astragalus Membranaceus Injection treatment. The expression of nuclear factor-kappaB (NF-kappaB) subunit p65 were evaluated by immuncytochemistical method, and the levels of p65 in the nuclei and the protein Ikappabetaalpha in the cytoplasm were evaluated by Western blotting. The levels of interleukin-6 (IL-6) and soluble intracellular adhesion molecule-1 (sICAM-1) in the cell culture were determined with ELISA.

RESULTSTNF-alpha induced the activation of NF-kappaB and increased the expressions of IL-6 and sICAM-1 in HUVECs. The activation of NF-kappabeta by TNF-alpha was suppressed by Astragalus Membranaceus Injection in a dose-dependent manner.

CONCLUSIONAstragalus Membranaceus Injection can inhibit the TNF-alpha-induced expression of IL-6 and sICAM-1 by suppressing NF-kappabeta activation, suggesting its protective effect on the endothelial function.

Astragalus membranaceus ; chemistry ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Cells ; cytology ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-6 ; metabolism ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors ; pharmacology ; Umbilical Veins ; cytology

Aged ; Coronary Angiography ; Coronary Artery Disease ; Coronary Disease ; diagnostic imaging ; pathology ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Tomography, Emission-Computed, Single-Photon

OBJECTIVETo study the in vitro antitumor activity of bcl-2 fully phosporothioated antisense oligodeoxynucleotide (bcl-2 ASODN) to malignant lymphoblastic cells.

METHODSProliferation and apoptosis of Raji cells incubated with bcl-2 ASODN were evaluated by MTT assay, flow cytometry (FCM) and electron microscopy, and the level of bcl-2 protein and mRNA expression were assessed by FCM and RT-PCR, respectively.

RESULTSMTT assay demonstrated that bcl-2 ASODN could partially inhibit the growth of Raji cells. After incubated with ASODN for 48 hours, Raji cells exhibited characteristic morphologic changes of apoptosis, including cytoplasm membrane blebbing, chromatin condensation crescents formation and nuclear fragmentation. The apoptosis rate of Raji cells treated with 20 micromol/L bcl-2 ASON for 72 hrs was 43.86% which is significantly higher than that of control (10.05%). The bcl-2 ASODN induced apoptosis of Raji cells was accompanied by declined expression of bcl-2 mRNA, which decreased to 0.88% at 72 hrs and was significantly lower than that of control (79.54%).

CONCLUSIONbcl-2 ASODN induced Raji cells apoptosis by downregulating bcl-2 protein.

Apoptosis ; drug effects ; Cell Division ; drug effects ; Dose-Response Relationship, Drug ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Oligonucleotides, Antisense ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Cells, Cultured ; drug effects ; metabolism

OBJECTIVETo investigate the complications of tongue base reduction with radiofrequency tissue ablation on patients with obstructive sleep apnea hypopnea syndrome (OSAHS) and find out the effective prevention strategies.

METHODSOne hundred and ninety three OSAHS patients diagnosed by polysomnography were received tongue base reduction with radiofrequency tissue ablation between March 2008 and December 2009. The intraoperative and postoperative complications including bleeding, hematoma of tongue base, abscess of tongue base, altered taste, tongue numbness, deviation of tongue extension movement, dysfunctions of pronunciation and swallowing as well as the managements were analyzed retrospectively.

RESULTSNo perioperative complications occurred. There were 186 cases with postoperative pain (96.4%), 155 cases with submandibular edema (80.3%). Nocturnal sudden cardiac death was encountered in 1 case and secondary bleeding in 1 case. There was no ulceration of tongue base mucose, hematoma or abscess of tongue base, altered taste, tongue numbness, tongue deviations, speech, swallowing and taste disorder after operation. The scale of postoperative pain claimed by patients was ranged between mild to moderate. Diclofenac suppository had analgesic effect for these patients. The quantity of bleeding in patient with secondary hemorrhage was so little that after proper treatment the bleeding was stopped and never happened again. Patient with nocturnal sudden cardiac death occurred at thirty-seven hours after operation, because of swelling and pain of tongue base aggravated sleep apnea and night hypoxemia inducing fatal arrhythmia.

CONCLUSIONSPostoperative pain and submandibular edema were 2 most common postoperative complications which can be easily controlled by antibiotics, Glucocorticoids and Diclofenac suppository. For those severe OSAHS patients accompanied by cardiopulmonary diseases, the tongue base reduction with radiofrequency tissue ablation can induce nocturnal sudden cardiac death. It is important to pay more attention on arrhythmias at night in severe OSAHS patients.

Adult ; Catheter Ablation ; adverse effects ; Edema ; pathology ; Female ; Humans ; Male ; Middle Aged ; Pain, Postoperative ; pathology ; Postoperative Complications ; Sleep Apnea, Obstructive ; surgery ; Tongue ; surgery

OBJECTIVETo study the effect of anti-HIV III B virus with extractions from Juglans regia, so as to searching for the new and efficacious leading compound of AIDS therapy.

METHODPhytochemical and chromatographical techniques were used to isolate compounds from J. regia; MT4 cells and HIV III B virus were used to study the effect of anti-HIV activity in vitro. BIACORE 3000 molecule coupled equipment were used for the target research also.

RESULTTwo extractions (B&E) were isolated from J. regia which possess the effect of anti-HIV activity. Targets study found that extraction B could affected on HIV-1 gp-41 fusing protein and extraction E could affected on HIV-1 integrase respectively.

CONCLUSIONJ. regia is a traditional Chinese medicine with active anti-HIV activity and worth to be studied further.

Cell Line ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; HIV-1 ; drug effects ; Humans ; Juglans ; chemistry ; Plant Extracts ; chemistry ; pharmacology

OBJECTIVETo assess the effect of choline in ameliorating lipopolysaccharide (LPS)-induced central nervous system inflammation and cognitive deficits in mice and explore the underlying mechanism.

METHODSSeventy-two mice were randomized into saline control group, LPS group, choline intervention group and choline control group. In the latter two groups, the mice received pretreatment with intraperitoneal injections of choline (40 mg/kg, 3 times daily for 3 consecutive days) prior to microinjection of LPS into the lateral cerebral ventricle to induce central nervous system inflammation; in saline and LPS groups, the mice were pretreated with saline in the same manner before intraventicular injection of artificial cerebrospinal fluid. Choline treatment was administered in the mice till the end of the experiment. The locomotor activity and spatial learning and memory capacity of the mice were examined. The expressions of Iba1 protein and proinflammatory cytokines (TNF-α and IL-β) I the hippocampal dentate gyrus, and the expressions of α 7nAchR, p38 MAPK and phosphorylated p38 MAPK in the hippocampus of the mice were detected.

RESULTSWater maze test showed that compared with the saline control group, the mice in LPS group exhibited significantly reduced platform crossings (P<0.05), which was significantly increased by choline pretreatment (P<0.05). The mice pretreated with LPS expressed obviously increased levels of IBA-1 protein, TNF-α, and IL-1β in the hippocampus (P<0.01), and choline pretreatment significantly lowered the expressions of IBA-1 protein and IL-1β (P<0.05). The phosphorylation level of p38 MAPK increased significantly after LPS pretreatment (P<0.05), and was reduced by choline pretreatment (P<0.05); α 7nAchR expression increased significantly in choline intervention group as compared with that in the other 3 groups (P<0.05).

CONCLUSIONCholine can probably antagonize LPS-induced hippocampal p38 MAPK phosphorylation in mice via the α 7nAchR signaling pathway to protective against LPS-induced neuroinflammation and cognitive impairment in mice.

Objective To compare the clinical efficacy of orthotopic ileal neobladder versus ortho- topic sigmoid neobladder.Methods The data of 96 patients who had undergone orthotopic ileal neoblad- der and 68 patients who had undergone orthotopic sigmoid neobladder were retrospectively analyzed.The perioperative condition,urinary continence,urodynamics,and pouch-related complications were compared between the 2 groups.Results Of all the 164 patients,12(7.3%)were lost to follow-up.The mean fol- low-up was 46(2-86)months in orthotopic ileal neobladder group,and 42(4-78)months in orthotopic sigmoid neobladder group.There was no significant difference in intraoperative blood loss and postoperative urinary continence between the 2 approaches(P＞0.05).However,compared with sigmoid neobladder group,ileal neobladder group had longer operative time and postoperative recovery time,and got a bigger pouch(P＜0.05).The early and late pouch-related complication rates of ileal neohladder group were 16. 7% and 29.2%,which were higher than those of sigmoid neobladder group.During the follow-up,tumor recurred in 3 cases of ileal neobladder group,but none in sigmoid neobladder group.Conclusions Ortho- topic ileal neobladder and sigmoid neobladder are similar in operative difficulties,and both can achieve satis- factory clinical results.Compared with ileal neobladder,sigmoid neobladder has shorter operative time, quicker recovery and lower rate of pouch-related complications,thus is a preferred procedure.