Objective To investigate the suppressive effect of carbon monoxide-releasing molecule Ⅱ (CORM-2) on LPS induced platelet α-granule exocytosis in sepsis via soluble N-ethylmaleimide-sensitive factor attached protein receptor/mammalian uncoordinated 18b (SNARE/Munc18b) complex formation.Methods Blood was collected from healthy volunteers' cubital vein, then platelets were isolated by differential centrifugation. Platelets were randomly divided into 5 groups. The control group did not undergo any treatment, the LPS group received 10 mg/L LPS simulation, the CORM-2 group and iCORM-2 group underwent LPS simulation and immediate administration of CORM-2 (10μmol/L and 50μmol/L) or iCORM-2 (50μmol/L), respectively. Samples were incubated in a CO2-incubator at 37 ℃, 95% humidity, and 5% CO2. Platelet α-granule contents were detected by using standard enzyme linked immunosorbent assay (ELISA), including platelet factor 4 (PF4), platelet derived growth factor-BB (PDGF-BB), and matrix metalloproteinase-2 (MMP-2). The expression of P-selectin was detected by flow cytometer. Transmission electron microscope and immunofluorescence microscope was used to assess platelet α-granules distribution. Expressions of Munc18b and SNARE proteins including vesicle-associated membrane protein-8 (VAMP-8), synaptosomal-associated protein-23 (SNAP-23) and syntaxin-11 (STX-11) were detected by Western Bolt. The SNARE/Munc18b complex formation was detected by immunoprecipitation.Results Compared with the control group, levels of PF4, PDGF-BB, MMP-2 and P-selectinin LPS-induced platelets were found to markedly elevated, while CORM-2 (10μmol/L and 50μmol/L) could decrease platelet α-granule contents exocytosis: [PF4 (μg/L): 7.69±0.58, 6.03±0.71 vs. 10.13±0.82; PDGF-BB (μg/L): 112.71±1.79, 102.91±5.86 vs. 128.78±1.39; MMP-2 (ng/L): 32.94±2.73, 27.58±3.36 vs. 53.26±1.21; P-selectin: (17.14±0.57)%, (15.35±0.68)% vs. (23.78±0.62)%; allP < 0.01]. Transmission electron microscope and immunofluorescence microscope showed that the extent of platelet α-granules assembled to platelet plasma membrane was significantly decreased following CORM-2 treatment. Compared with the control group, the expressions of Munc18b and SNARE proteins and SNARE/Munc18b complex formation in LPS-stimulated platelets were significantly increased, while CORM-2 (10μmol/L and 50μmol/L) inhibited these elevations (Munc18b/GAPDH: 0.80±0.08, 0.69±0.01 vs. 0.99±0.09; VAMP-8/GAPDH: 0.72±0.09, 0.50±0.12 vs. 1.18±0.14; SNAP-23/GAPDH: 1.18±0.22, 0.63±0.10 vs. 1.90±0.08; STX-11/GAPDH: 0.76±0.02, 0.57±0.08 vs. 1.16±0.23; VAMP-8/ Munc18b: 0.65±0.09, 0.53±0.07 vs. 1.21±0.20; SNAP-23/Munc18b: 0.85±0.07, 0.55±0.09 vs. 1.26±0.08; STX-11/ Munc18b: 0.78±0.05, 0.61±0.10 vs. 1.39±0.16; allP < 0.01). Above all, the data showed a dose dependent change.Conclusion We could suggest that CORM-2 suppressed α-granule exocytosis in LPS-stimulated platelets and the potential mechanisms might involve SNARE/Munc18b complex formation.

Objective To investigate the effect of the cerebral infarction in rats and the role of matrix metalloproteinases-2 (MMP-2), matrix metalloproteinases-9 (MMP-9), and tissue inhibitor of metalloproteinases-3 (TIMP-3) in the pathogenesis. Method Sixty-six rats were randomly divided into 3 groups: blank group (n= 6), control group (n=30), model group (n=30). The rat model was established by carotid injection of auto blood thrombus, the control group was given the operation without injection. Each group was subdivided into 24 hours, 48 hours, 3 days, 5 days, 7 days after operative intervention. At serial intervals, the animals in each group were sacrificed and brain tissue samples were collected to determine MMP-2, MMP-9, TIMP-3 expression by immunohistoehemistry. The changes of behaviors were observed and the histopathological changes were investigated by hematoxylin and eosin stain. Results The experimental group had more obvious behavior change than other groups. Hematoxylin and eosin stain showed significant histopathological changes; MMP-2, MMP-9 and TIMP-3 expression was up-regulated in comparison with other groups (P

Objective To observe the degeneration of Brugia malayi in Meriones unguiculatus model. Methods Microfilaria of Brugia malayi derived from Meriones unguiculatus was used to infect Anopheles sinensis . Infective stage larvae (L 3) from mosquito vector were collected and inoculated into abdomen of Meriones unguiculatus. Successive 33 generations of the parasite in the rodent model have been observed. Results Since 1974 when the animal model was established, the parasite has lasted for 33 generations, the positive rate of Meriones unguiculatus with microfilaria gradually reduced from 80% of the 28th generation to 16% of the 32nd generation and finally to 0 at the 33rd generation. Conclusion It became difficult for the larvae of Brugia malayi to develop and/or reproduce in the animal model after multiple inoculations for generations.

Porcine Reproductive and Respiratory Syndrome virus (PRRSV) is the etiological agent of Por- cine Reproductive and Respiratory Syndrome. We summarized the recent research progress on molecular bi- ology of PRRSV including the structure of genome, viral structural and Non-structural protein.

Objective To provide reference for clinical pharmacists participating in pharmacotherapy of low molecular weight heparin calium-induced thrombocytopenia.Methods Clinical pharmacists carried out pharmaceutical care for a patient with chronic cor pulmonale with acute exacerbation of chronic asthmatic bronchitis in the use of low molecular weight heparin calium,who developed progressive thrombocytopenia,and helped clinicians to manage adverse reaction and select subsequent treatment drug.Results The suggestions were adopted by clinicians.The platelets of patients gradually recovercd without thrombosis events.Condusion Participation of clinical pharmacists in the treatment of low molecular weight heparin calium induced-thrombocytopenia can effectively improve the prognosis of patients and ensure the safety in drug use.

Background Acanthamoeba keratitis is a sort of serious infectious eye disease with high causing-blindness rate.Acanthamoeba keratitis often is misdiagnosed as fungal keratitis or viral keratitis in the early stage.Because conventional clinical diagnosis methods show a low specificity and take a long time,timely treatment often is delayed.Conventional PCR does not apply well because the lesion sample is not enough to extract DNA.However,direct PCR can amplify 18S rRNA conserved sequence of acanthamoeba keratitis without the extraction of DNA.Objective This study was to discuss the feasibility for rapid diagnosis of acanthamoeba keratitis using direct PCR to amplify the gene 18S rRNA fragment.Methods Ten acanthamoeba strains were isolated from 10 eyes with acanthamoeba keratitis in Qingdao Eye Hospital.The sensitivity of the direct PCR assay was tested using different numbers of amoebas.The specificity of the assay was tested using DNA extracted from acanthamoeba,candida albicans,pseudomonas aeruginosa,herpes simplex virus-1 (HSV-1) and normal human corneal epithelial cell.Acanthamoeba keratitis models were established using infected method in clean 6-week-old female BALB/c mice.Corneal lesion samples were obtained 1 day,3,5,7,10,15 days after modeled.The effectivity and feasibility of the direct PCR assay for rapid diagnosis of acanthamoeba keratitis were evaluated and compared with culture method,corneal smear examination and real-time PCR.Results Direct PCR primers could only amplify DNA of acanthamoeba rather than other pathogens,and 10 stains of acanthamoeba were detected at least in each sample.During the development of acanthamoeba keratitis in the mice,the diagnosis positive rate of direct PCR was 80.0％,90.0％,80.0％,70.0％,70.0％ and 50.0％ in 1 day,3,5,7,10,15 days after modeled with the total positive rate 73.3％,which was higher than 31.7％ of culture method,56.7％ of corneal smear examination and 61.7％ of realtime PCR,with a significant difference between the direct PCR and culture method (P =0.005),but no significant difference was seen in the total positive rate between the direct PCR and real-time PC R (P =0.172) or corneal smear examination (P =0.056).Conclusions The direct PCR assay is a simple,rapid,highly specific and sensitive method for the rapid diagnosis of acanthamoeba keratitis,especially for the limited lesion sample.

Objective To evaluate the effect of deep and moderate neuromuscular blockade on surgical conditions during minor laparoscopic gynecologic surgery.Methods Sixty-five patients,with expected surgery time ＜ 3 h,aged 18-60 yr,with body mass index＜30 kg/m2,of American Society of Anesthesiologists physical status Ⅰ orⅡ,scheduled for elective laparoscopic gynecological surgery,were allocated into deep neuromuscular blockade group (group D,n =33) and moderate neuromuscular blockade group (group M,n=32) using a random number table.After induction of anesthesia,the patients were tracheally intubated and mechanically ventilated.Cisatracurium was continuously infused to maintain the degree of neuromuscular blockade in both groups to achieve the target degree post-tetanic count of 1 or 2 in group D and train-of-four (TOF) count of 1 or 2 in group M.Surgical conditions were assessed and scored after surgery.The recovery index,time for TOF ratio returning to 0.7 and 0.9,surgery time,mean intra-abdominal pressure,extubation time and TOF ratio at extubation were recorded.Results Compared with group M,the mean intra-abdominal pressure was significantly decreased,and the extubation time and time for TOF ratio returning to 0.7 and 0.9 were prolonged in group D (P＜0.05).There was no significant difference in the other parameters between the two groups (P＞0.05).Conclusion Moderate neuromuscular blockade can provide better surgical conditions for minor laparoscopic gynecological surgery with shorter recovery time.

ObjectiveTo investigate the clinical feature peripheral neuropathy and vasculopathy after acrylamide toxication. Methods2 young male patients with peripheral neuropathy who had exposed to acrylamide for job more than one year were reported.ResultsNeuroelectrophysiological examination showed marked abnormalities in both peripheral and central nerve conduction in both patients. Sural nerve biopsies revealed axonal degeneration, Wallerian degeneration and giant axon with accumulated neurofilaments. Additonally, vasculopathies including prominant thickness of arterial intesma and basal membrane of capillary as well as apoptosis of vascular pericyte, were evident. ConclusionAxonal degeneration and vascular involvement has been found in acrylamide toxication. Vascular impairment maybe plays an important role in the pathogenesis of neuropathy.

This retrospective analysis compared standard regimen of doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) with the dose-dense ABVD regimen (ABVD-21) in terms of efficacy and toxicity. Patients who had early-stage unfavorable or advanced Hodgkin's lymphoma (HL) according to German Hodgkin Study Group criteria from March 1999 to February 2011 were analyzed for treatment response, long-term survival and hematological toxicity. There were 85 patients in the ABVD-21 group and 118 patients in the ABVD group respectively. The complete remission rates after completion of treatment were 92.9% and 90.7% for ABVD-21 and ABVD, respectively. During a median follow-up period of 62 months, no significant difference was found in projected 10-year progression-free survival (PFS) and overall survival (OS) rates (84.7% and 94.1% respectively for ABVD-21; 81.4% and 91.5% for ABVD). Subgroup analyses showed that ABVD-21 was significantly better than ABVD for patients with IPS≥3 in terms of PFS and OS rates. Grade 3 to 4 leukopenia (51.8% vs. 28.8%, P=0.001) and neutropenia (57.6% vs. 39.0%, P=0.009) were more common with ABVD-21. We were led to conclude that dose-dense ABVD did not result in better tumor control and overall survival than did ABVD for early-stage unfavorable HL. However, patients at high risk, for example, with IPS≥3, may benefit from dose-dense ABVD.

BACKGROUND:Intervertebral disc degeneration is one of the ancient and common clinical diseases. Its complex pathogenesis affected by various factors, such as environment and genes, is stil in debate. Because of the technical limitations, there is stil no deep understanding on the molecular mechanism of intervertebral disc degeneration. However, its molecular mechanism in recent years has made considerable development. OBJECTIVE: To summarize and discuss the molecular mechanism of intervertebral disc degeneration, thereby providing the basis for the effective treatment. METHODS: CNKI and Medline databases were retrieved by the first author using computer to search relevant articles published from 2005 to 2015. The key words were “intervertebral disc degeneration, molecular mechanism, environmental factors, genes, matrix, degradation enzyme,inflammatory factor, biological environment, treatment” in Chinese and English, respectively. Mechanisms of intervertebral disc degeneration, involving genes, cel senescence and apoptosis, degradation enzyme and substrate, inflammatory cytokines, were summarized to explore the pathogenesis and possible effective treatment of intervertebral disc degeneration. RESULTS AND CONCLUSION:Totaly 153 articles were initialy retrieved and finaly 52 articles were included in result analysis according to inclusive and exclusive criteria. Unique structure and biochemical properties of the intervertebral disc are easy to cause intervertebral disc degeneration. Traditionaly, environmental factors, such as occupation and smoking, are considered as the main factors inducing intervertebral disc degeneration; however, more and more studies have shown that genes have the most important influence on intervertebral disc degeneration. Declined extracelular matrix, increased degradation enzymes, and overexpression of inflammatory factors can al destroy the entire structure of intervertebral disc, and accelerate the process of intervertebral disc degeneration. Effective treatment for intervertebral disc degeneration can be formulated depending on the deep understanding on its molecular mechanisms. Although there is a further understanding on the molecular medium of intervertebral disc degeneration, the complex biochemical environment within the intervertebral disc is stil a great chalenge to the treatment of intervertebral disc degeneration.