Objective To investigate whether N-methyl-D-aspartate(NMDA) signal pathway is involved in platelet-activating factor(PAF)-induced apoptosis in neurons.Methods Mice cortex neurons were treated by PAF in different dose for 24 hours or pretreated by a PAF receptor antagonist BN52021, an NMDA receptor antagonist MK801 and a nitric oxide synthase(NOS) inhibitor L-NAME for 30 min. Then the neurons were stained by propidium iodide(PI)/calcein AM and the images were taken by a digital camera on a fluorescent microscope. Neuron death ratio(%) was calculated. In the mean time, neuron NOS(nNOS) expression of the cells was detected by Western Blot method, as well as nNOS activity was measured by 3H radioimmunoassay.Results (1)Neuronal death increased after neurons were treated with 0.01, 0.1, 0.3 and 0.6 ?mol/L PAF 24 h. There were signifitantly changed in 0.3 ?mol/L and 0.6?mol/L comparing control. (2)PAF-induced neurotoxicity was prevented not only by BN5021, but also by MK-801 or L-NAME. (3)PAF enhanced nNOS expression and activity in neurons.Conclusion NMDA signal pathway is involved in platelet-activating factor-induced neurotoxicity.

ve To investigate the effect of ischemic preconditioning (IPC) on apoptosis induced by acute myocardial ischemia/reperfusion and expressions of Bcl-2 and Bax protein which were known to modulate apoptosis. Methods Twenty-four healthy SD rats of either sex, weighing (200()20)g were anesthetized with intraperitoneal pentobarbital 4.5mg?100g-1. The animals were tracheotomized and mechanically ventilated. Respiratory rate was 20 bpm and tidal volume 2 ml?100g-1. Myocardial ischemia/ reperfusion(I/R) model was established by ligation and untying of the anterior descending branch of left coronary artery. The animals were randomly divided into three equal groups with eight animals each. Group I : (control group) anterior descending branch was exposed and dissected but not ligated and was exposed for 50 min. Group II (I/R group): anterior descending branch was double ligated for 30 min and then untied for reperfusion which lasted 2h. Group III (IPC group): The anterior descending branch was tied for 5 min then untied for 5 min and the process was repeated 4 times according to Murray's method, then I/R was produced as in group II. A piece of myocardium of 2 mm thick was cut from ischemia-infarct area. Apoptotic myocardial cells were detected by TdT-mediated dUTP-biotin nick end labeling (TUNEL) and the expressions of Bcl-2 and Bax protein were measured by immunohistochemical technique. Results I/R increased the percentage of apoptotic myocardial cells and the optical density (OD) value of Bax protein and decreased the OD value of Bcl-2 protein as compared with those in the control group. IPC reduced the increased percentage of apoptotic myocardial cells and OD value of Bax protein induced by I/R and increased the OD value of Bcl-2 protein as compared with those in the I/R group. Conclusions IPC can inhibit the apoptosis induced by myocardial I/R by modulating the expression of Bcl-2 and Bax protein.

Objective To study a safe means of mandibular angle plasty of bone cutting and its clinical efficay.Methods Through grinding bone oblique trapezoidal incisure,mandibular angle wide deformity was then corrected according to the cutting trace on bone cutting method.Results Of 260 cases of torture beauty,185 cases were followed-up,in which 181 (97.84％) cases were satisfied with the results,and 1 case got satisfaction after facial liposuction; other 1 case did not accepted further treatment advice of bone cutting processing because her mandibular angle was too wide.Two cases of bone wax reaction occured and got satisfaction after treatment.Conclusions Mandibular angle plasty by combined grinding with cutting makes it easy for positioning and bone cutting; bone cutting arc becomes more fluent with good exposure of its posterior horn.It can prevent mandibular fracture induced by bone cutting that does not reach the designated position.Therefore,this procedure is relatively simple and safe.

Objective:To explore the relationship between the nitric oxide(NO)and the pathogenesis of the pulmonary hypertension(PH)induced by congenital heart disease(CHD). Method:NO content in plasma of CHD patient's pulmonory artery and superior vena cava was detected with NO kit. Result:NO content in the pulmonary artery of the patients with PH was much higher than that of patients without PH(37.58?9.99?mol/L vs 19.03?15.25?mol/L,P0.5). Conclusion:NO content in the pulmonary artery of CHD patients with PH increase,which may be involved in the pathogenesis of PH induced by CHD.

To investigate the clinical characteristics and angio-architecture features of patients with dural arteriovenous fistulas (DAVF).The clinical data of 48 patients with DAVF were analyzed retrospectively.All patients were diagnosed by digital subtract angiography and 43 cases were also examined by MRI.Patients were divided into the bleeding group and non-bleeding group,whose clinical features and angio-architecture were compared.Of 48 cases,13 patients demonstrated intracranial bleeding,and men were more common in bleeding group (M/F:10/3) than in non-bleeding group (M/F:15/20) (P =0.036).The Cognard scores of bleeding group and nou-bleeding group were 3.77 ±0.28 and 2.49 ±0.21,respectively (P =0.002) ; however,there was no significant difference in age and the number of feeding artery between two groups.The results indicate that male patients with high Cognard scores tend to intracranial bleeding.

The amino acid sequence of the fusion protein FP3 was measured by two types of LC-MS/MS and its primary structure was confirmed. After reduction and alkylation, the protein was digested with trypsin and glycosyl groups in glycopeptide were removed by PNGase F. The mixed peptides were separated by LC, then Q-TOF and Ion trap tandem mass spectrometry were used to measure b, y fragment ions of each peptide to analyze the amino acid sequence of fusion protein FP3. Seventy-six percent of full amino acid sequence of the fusion protein FP3 was measured by LC-ESI-Q-TOF with the remaining 24% completed by LC-ESI-Trap. As LC-MS and tandem mass spectrometry are rapid, sensitive, accurate to measure the protein amino acid sequence, they are important approach to structure analysis and identification of recombinant protein.

The study aims to characterize and compare interferon reference standards from 5 manufacturers. By testing molecular mass and trypsin-digested peptide mass mapping, the amino acid sequence was verified and post-translational modifications such as disulfide bond were identified. Results show that the molecular mass and amino acid sequence were consistent with theory; the disulfide bonds of 4 lots of interferon were Cys1-Cys98/Cys29-Cys138, 1 lot was Cys29-Cys139/Cys86-Cys99; N-terminal "+Met", acetyl N-terminal and Met oxidation were identified in part of the sample. UPLC-MS can be used to characterize and compare interferon reference standards from different manufacturers.

Recombinant human interferon α2b(rhIFNα2b)is widely used as an antiviral therapy agent for the treatment of hepatitis B and hepatitis C.The current identification test for rhIFNα2b is complex.In this study,an anti-rhIFNα2b nanobody was discovered and used for the development of a rapid lateral flow strip for the identification of rhIFNα2b.RhIFNα2b was used to immunize an alpaca,which established a phage nanobody library.After five steps of enrichment,the nanobody I22,which specifically bound rhIFNα2b,was isolated and inserted into the prokaryotic expression vector pET28a.After subsequent purification,the physicochemical properties of the nanobody were determined.A semiquantitative detection and rapid identification assay of rhIFNα2b was developed using this novel nanobody.To develop a rapid test,the nanobody I22 was coupled with a colloidal gold to produce lateral-flow test strips.The developed rhIFNα2b detection assay had a limit of detection of 1 μg/mL.The isolation of I22 and successful construction of a lateral-flow immunochromatographic test strip demonstrated the feasibility of performing ligand-binding assays on a lateral-flow test strip using recombinant protein products.The principle of this novel assay is generally applicable for the rapid testing of other com-mercial products,with a great potential for routine use in detecting counterfeit recombinant protein products.

BACKGROUND:Studies have shown that posterior cruciate ligament injuries are associated with the anatomical morphology of the knee joint. OBJECTIVE:To explore anatomical morphological risk factors for posterior cruciate ligament injury. METHODS:The imaging data of 142 patients who visited Affiliated Hospital of Binzhou Medical University for knee joint problems from January 2015 to August 2022 were retrospectively analyzed.They were divided into posterior cruciate ligament injury group(n=71,including 49 males and 22 females)and posterior cruciate ligament intact group(n=71,including 49 males and 22 females).Intercondylar notch width,intercondylar notch height,bicondyle width,notch width index,angle of intercondylar notch,Blumensaat's line inlication angle,medial tibial spine height,lateral tibial spine height,tibial spine width,tibiofemoral consistency index,tibial plateau anterior-posterior diameter,medial tibial depth and patellar tendon-tibial shaft angle were measured on MRI images.Posterior tibial slope was measured on X-ray images.The above indicators were included in the logistic regression analysis for investigation. RESULTS AND CONCLUSION:(1)Univariate logistic regression analysis in men showed that tibial spine width,tibiofemoral consistency index,medial tibial depth,and posterior tibial slope were associated with posterior cruciate ligament injury(P<0.05).Multivariate binary logistic regression analysis showed that tibiofemoral consistency index and medial tibial depth were associated with posterior cruciate ligament damage(P<0.05).(2)Univariate logistic regression analysis in women showed that medial tibial spine height,lateral tibial spine height,tibial spine width,and posterior tibial slope were associated with posterior cruciate ligament injury(P<0.05).Multivariate binary logistic regression analysis showed that posterior tibial slope was associated with posterior cruciate ligament damage(P<0.05).(3)The receiver operating characteristic curve showed that tibiofemoral consistency index,medial tibial depth and posterior tibial slope had a certain predictive value on posterior cruciate ligament damage.(4)These findings suggest that anatomical morphological risk factors for posterior cruciate ligament injury differ between men and women,and tibial spine width and posterior tibial slope are common risk factors.In the male population,abnormal tibial spine width,tibiofemoral consistency index,medial tibial depth,and posterior tibial slope are easy to induce posterior cruciate ligament injury.In the female population,abnormal medial tibial spine height,lateral tibial spine height,tibial spine width,and posterior tibial slope are easy to induce posterior cruciate ligament injury.Clinicians can use the above risk factors to identify abnormal knee morphology,assess people at risk of posterior cruciate ligament injury,and provide preventive advice and guidance for treatment.