OBJECTIVETo explore the neuroprotective effect and mechanism of picroside II on extracellular regulated protein kinases1/2 (ERK1/2) signal transduction pathway in cerebral ischemia injuryrats. METHODS The middle cerebral artery occlusion (MCAO) model was established by inserting a monofilament into middle cerebral artery. Totally 96 successfully modeled Wistar rats were divided into the modelgroup, the treatment (picroside II) group, the Lipopolysachcaride (LPS) group, and the U0126 group according to random digit table. Each group was further divided into 3 subgroups, i.e. 6, 12, and 24 h sub-groups. Picroside II (20 mg/kg) was peritoneally injected to rats in the treatment group 2 h after ischemia.LPS (20 mg/kg) and Picroside II (20 mg/kg) were peritoneally injected to rats in the LPS group 2 h after ischemia. U0126-EtOH (20 mg/kg)and Picroside II (20 mg/kg) were peritoneally injected to rats in the U0126group 2 h after ischemia. Equal volume of normal saline was peritoneally injected to rats in the control groupand the model group. The neurobehavioral function was evaluated by modified neurological severity score(mNSS) test. The structure of neurons was observed using hematoxylin-eosinstaining (HE) staining. Theapoptotic cells were detected using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The expression of phosphorylated extracellular signal-regulated protein kinase1,2 (pERK1,2) in cortex was detected using immunohistochemistry (IHC) and Western blot.

RESULTSAfter cerebral ischemia injury neurological impairment score increased, the damage of neuron in the cortical area was aggravated, apoptotic cells increased in the model group as time went by. The expression of pERK1/2 increased more significantly in the model group than in the control group (P <0.05). The damage of neuron in the cortical area was milder, while apoptotic cells decreased, the expression of pERK1f2 obviously decreased more in the treatment group and the U0126 group (P < 0.05). The early damage of neuron in the cortical area was more severe, apoptotic cells and the expression of pERK12 were comparatively higher in early stage of the LPS group, but the expression of pERK1/2 was somewhat decreased in late stage.

CONCLUSIONSActivating ERK12 pathway could mediate apoptosis and inflammatory reactions of neurons after cerebral ischemia injury. Picroside II could protect the nerve system possibly through reducing activation of ERKI2 pathway, inhibiting apoptosis of neurons and inflammation reaction.

Animals ; Apoptosis ; Brain Ischemia ; drug therapy ; Cinnamates ; pharmacology ; Infarction, Middle Cerebral Artery ; drug therapy ; Iridoid Glucosides ; pharmacology ; MAP Kinase Signaling System ; drug effects ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar

·α-crystallin is the predominant structural protein in the lens.lt is a member of small heat shock proteins ( sHSPs) which has the common functions of HSPs.lt also has anti-apoptotic activity etc.Recently, it has been proved to combine with the cellular membrane of retinal ganglion cells ( RGCs ) to enhance the survival of RGCs and the regeneration of axons, thereby partly restore visual function.But we haven’t come to a unified conclusion of the mechanism.This review is focused on structure and functions of α-crystallin, the protection function and mechanism of α-crystallin towards RGCs after the optic nerve injury.

Carrier Proteins ; Humans ; Inflammasomes ; NLR Family, Pyrin Domain-Containing 3 Protein ; Silicosis ; pathology

Objective To explore role of 64-slices spiral CT in differetiation of acute chest pains.Methods Thirty six patients with acute chest pains were performed 64-slices spiral CT chest angiography.Two-dimensional and three-dimensional reconstruction was performed in all patients by means of multiplanar reconstruction(MPR)(coronal,sgittal oblique),curved planar reformation(CPR), maximum intensity projection(MIP),and volume rendering(VR).All images were blindly reading by two experienced radiologist.DSA were performed at the same time in 16 cases.Results The coronary artery branches,pulmonary artery and aortic artery in all patients were showed clearly,The acute myocardial infarction were showed in 10 cases,The pulmonary artery embolism in 14 cases,The aortic dissection in 6 cases respectively,The Coronary embolism in One case ,pneumothorax In One case The constrictive pericarditis in 1 case respectively.Normal findings in 4 cases.Conclusion 64-slices spiral CT is a useful and noninvasive examination in acute chest pain.

In this paper, fourteen new L-proline derivatives were designed and synthesized, and their acetlcholinesterase (AChE) inhibitory activities were also investigated in vitro. New L-proline derivatives were prepared from substituted 2-bromo-1-acetophenones through four-step reaction; and their bioactivities as AChE inhibitors were measured by Ellman spectrophotometry. The results showed that the target compounds had a certain AChE inhibitory activity to in vitro. The bioactivity of compound 8b was the best of them, and its IC50 value was 5.45 µmol.L-1, which was better than that of rivastigmine. So the acetylcholinesterase inhibitory activities of new L-proline derivatives were worth to be further studied.
Acetylcholinesterase ; Cholinesterase Inhibitors ; chemical synthesis ; chemistry ; Drug Design ; Proline ; analogs & derivatives ; Rivastigmine ; chemistry ; Structure-Activity Relationship

Objective To investigate the feasibility and clinical effectiveness of interventional ventricular septal defect(VSD) occlusion using asymmetric occluders in patients whose VSD are adjacent to aortic valves.Methods One hundred and fifty-two patients from our hospital whose VSD were occluded with asymmetric occluders were examined by echocardiography.Dimensions,shapes and shunts of VSD,structures around defects were observed and aortic valve prolapse(AVP) degrees were accessed before the interventions.The patients were followed up periodically after the procedures.The longest follow-up was 6 years.Results Sizes of VSD measured from left ventricular side were larger than those measured from right ventricular side.Twenty-seven cases with VSD had mild AVP.Right sides of defects had adhesions with variable morphological patterns in 85% patients with perimembranous VSD.After occlusion,there were 7 cases showed residual shunt,2 cases with the third degree atrio-ventricular blockages,3 cases appeared aortic-valve regurgitations (AR) and 1 case with slightly increased AR.Conclusions Immediate and medium-term effects of interventional occlusion with asymmetric occluders were satisfactory in the patients with VSD and mild AVP whose aortic valve margin were insufficient to symmetric occluders.Surgical operation should be preferred to patients with VSD and moderate to severe AVP and to very young children.Pre-closure echocardiographic evaluation is critical to the success of interventional closure with asymmetric occluder and the long-term out coming.

Abstract：Objective To investigate the morphological features of the Pneumocystis jirovecii, in order to facilitate early detection and rapid diagnosis of this rare pathogen from a morphology point of view by laboratory technicians. By analyzing the laboratory features and application value of different pathogen detection methods in the diagnosis of Pneumocystis jirovecii pneumonia, we aim to provide the most reliable diagnostic basis for rapid diagnosis of Pneumocystis jirovecii pneumonia.Methods A retrospective analysis was conducted on the test results of bronchoalveolar lavage fluid samples from a comprehensive hospital in Zhangqiu District, Jinan City, Shandong Province, and a hospital in Changde City from April 2022 to October 2022. Five confirmed cases of Pneumocystis jirovecii pneumonia were detected. Its clinical manifestations, laboratory results, and morphological characteristics of pathogens under different stains were analyzed to discuss the advantages and disadvantages of different detection methods. Results Cytological examination of bronchoalveolar lavage fluid found the trophozoites and cysts of Pneumocystis jirovecii by Wright's-Giemsa staining in 4 cases (80%), and the cysts of Pneumocystis jirovecii by Silver hexamine staining in 4 cases (80%), while the metagenomic next-generation sequencing confirmed all the 5 positive results. All 5 patients had different degrees of reduction in the absolute count of peripheral blood lymphocytes, and the serum lactic dehydrogenase and (1-3)-β-D-Glucan were increased. Among the 5 patients in this study, 4 were treated with sulfamethoxazole combined with caspofungin, and 1 was treated with sulfamethoxazole. Three patients were cured and discharged from hospital after treatment, but two died. Conclusions The method of Wright's-Giemsa staining for the cytological examination of bronchoalveolar lavage fluid to find Pneumocystis jirovecii has the unique and irreplaceable advantages as silver staining. Metagenomic next-generation sequencing can further increase the positive detection rate of Pneumocystis jirovecii. The combination of cytological examination of bronchoalveolar lavage fluid with metagenomic nextgeneration sequencing is a powerful diagnostic method for rapid diagnosis of Pneumocystis jirovecii pneumonia, which can diagnose accurately and reduce missed diagnosis.

BACKGROUND: The ideal biomaterial means absence of cytotoxic effect and immunological rejection, degradation at right moment, and a well histocompatibility. Whether bio-derived bone can be used in vivo for long time and exerts functions deserves to be studied.OBJECTIVE: To investigate the local histocompatibility after bio-derived bone implanted into mouse and the effect on immunofunctions.DESIGN: A randomized controlled animal experiment. SETTING: Key Laboratory of Pathobiology, Ministry of Education, School of Basic Medical Sciences, Jilin University. MATERIALS: This study was performed at the Key Laboratory of Pathobiology, Ministry of Science, School of Basic Medical Sciences, Jilin University from March to July in 2006. Eighteen BALB/C mice (weighing 20±2 g, half male and half female), one male Kunming mouse (weighing 20 g), and one female rabbit (weighing 2.5 kg) were included in the experiment. All the experimental animals were provided by Laboratory Animal Center, School of Basic Medical Science, Jilin University. The disposal of the experimental animals in the test process accorded with ethical guidelines for the use and care of animals. Porcine cancellous bone (iliac bone) was purchased from the market. Iscove's Modified Dulbecco's Medium (IMDM, Hycolone, USA), fetal bovine serum (FBS, Gibco Co., Ltd, USA), methyl thiazolyl tetrazolium (MTT, Sigma, USA), and concanavalin A (ConA, Sigma Co., Ltd, USA) were used.METHODS: Bio-derived bone was prepared from commercial porcine bone. ① Eighteen BALB/C mice were randomly divided into three groups with 6 mice in each group: a control group (simple local muscle injury without implantation), a bio-derived bone implantation group ( implanting bio-derived bone into the lower limb), and a xenogenic bone implantation group (femoral bone from Kunming mouse was implanted into the muscle of lower limb). ② Twenty-one days after operation, the implant material and surrounding tissue were obtained for gross observation and haematoxylin-eosin staining to investigate the histocompatibility of bio-derived bone. Mouse immunofunction was assessed by complement-mediated cytotoxicity test. Absorbance was determined with an automatic ELISA reader at 570 nm to assess the cytotoxicity.MAIN OUTCOME MEASURES: ①Histocompatibility of implant surrounded tissue. ②Lymphocyte stimulation indices after induction of concanavalin A. ③ Cytotoxicity in each group after complement-dependent cytotoxicity test.RESULTS: Eighteen BALB/C mice were included in the final analysis. ①Histocompatibility of implant surrounded tissue: In the bio-derived bone implantation group, 21 days after bio-derived bone implantation, there were no presentation of congestion, degeneration, necrosis and diapyesis around the implant in gross, plenty of fibrous connective tissue invaded into the pores of the bio-derived bone, encapsulation and forming the fibrous capsule. A great quantity of neutrophils and macrophages were not detected around the implant by haematoxylin-eosin staining. Bio-derived bone was encapsulated with fibrous tissue, and part of the biomaterial began to degrade, and being replaced with fibrous tissue. Regarding xenogenic bone implantation group, necrotic tissue was detected in the cross-section of the muscle in gross. A lot of neutrophils, macrophages and necrotic tissue were detected around the implant by haematoxylin-eosin staining. ②Lymphocyte stimulation indices: The stimulation index of xenogenic bone implantation group was significantly larger than that of control group (P ＜ 0.05). There was no statistical difference between the bio-derived bone group and the control group (P ＞ 0.05). ③Cytotoxicity: The cytotoxicity of xenogenic bone implantaion group was significantly larger than that of control group (P ＜ 0.05). There was no significant difference in cytotoxicity between the bio-derived bone implantation group and the control group (P ＞ 0.05).CONCLUSION: The obtained bio-derived bone causes little immunoreactions, has no obvious cytotoxicity or inflammatory reactions, and possesses a good histocompatibility and bio-safety.

ObjectiveTo investigate the efficacy of co-infusing cord blood (CB) as the third party cells on graft versus host disease (allo-GVHD) prophylaxis after unrelated or haploidentical donor allogeneic hematopoietic stem cell transplantation (allo-HSCT).MethodsFrom 2007 to 2011,41 patients receiving unrelated or haploidentical donor allo-HSCT were analyzed retrospectively.Twenty-five patients received one unit of HLA 4/6-6/6 matched CB one day before SCT as CB group,and median MNC dose was (1.64 ± 0.49) × 107/kg.Sixteen cases not receiving CB served as control group.All patients received antithymocyte globulin,cyclosporine,methotrexate,and mycophenolate mofetil as GVHD prophylaxis.The incidence and severity of aGVHD,and treatment-related mortality were compared between two groups.ResultsThe main clinical characteristics in both groups were comparable.The cumulative incidence of aGVHD in CB group and control group was 44.0％ versus 68.8％ respectively (x2 =2.403,P＞0.05).The cumulative incidence of grades Ⅲ to ⅣV aGVHD in CB group and control group was 16.0％ and 37.5％ respectively (x2 =2.445,P＞0.05).The 100-day treatment-related mortality in CB group and control group was 12.0％ and 12.5％ respectively (x2 =0.002,P＞0.05).ConclusionCord blood as the third party cells might reduce the incidence and severity of aGVHD in unrelated or haploidentical donor HSCT.The efficacy and the mechanism of this strategy need to be further explored by prospective randomized controlled trials.

This study was designed to analyze the effect of the mitochondrial respiratory pathways of Candida albicans (C.albicans) on the biofilm formation.The 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay was used to measure the metabolic activities of biofilms formed by the C.albicans which were cultured in the presence of respiratory pathways inhibitors.The biofilms formed by the wide type (WT),GOA1-deleted (GOA31),GOA1-reconstituted (GOA32),AOX1a-deleted (AOX1) and AOX1b-deleted (AOX2) C.albicans strains were examined by the XTT reduction assay and fluorescence microscopy.The expression of adhesion-related genes BCR1,ALS1,ALS3,ECE1 and HWP1 in the biofilms formed by the above five C.albicans strains was detected by real time polymerase chain reaction.It was found that the metabolic activity of biofilms formed by C.albicans was decreased in the presence of alternative oxidase inhibitor whereas it was increased in the presence of classical mitochondrial respiratory pathway complex Ⅲ or complex Ⅳ inhibitor.AOX1 strain produced scarce biofilms interspersed with few hyphal filaments.Moreover,no significant changes in the expression of BCR1 and ALS3 were observed in the AOX1 strain,but the expression of ALS1 and ECE1 was down-regulated,and that of HWP1 was up-regulated.These results indicate that both AOX1 and AOX2 can promote the biofilm formation.However,AOX1a primarily plays a regulatory role in biofilm formation in the absence of inducers where the promoting effect is mainly achieved by promoting mycelial formation.