OBJECTIVE: To investigate the relation between the expression of tPA, MMP-2, MMP-9 and AEG-1 in the human brain tissue and the ethanol concentration under the acute alcohol poison, and to analyze the role of alcohol and trauma in the mechanism of death of subarachnoid hemorrhage. METHODS: Fifteen real cases were collected in this study. The brain tissues were researched by histological examination and the concentration of ethanol in heart blood were detected. The tPA, MMP-2, MMP-9 and AEG-1 in brainstem, brain and cerebellum were observed respectively by immunohistochemistry. RESULTS: In alcohol poisoning groups with or without trauma, the acute alcohol toxicity resulted in the swelling of brain tissues. The tPA, MMP-2, MMP-9 and AEG-1 of brainstem, brain and cerebellum showed high expression in alcohol victims, and the tPA in cerebellum showed no difference. The expression of the MMP-2, MMP-9 and AEG-1 showed good relation with the ethanol concentration in blood (P < 0.05, r > 0.6). CONCLUSION The expressions of tPA, MMP-2, MMP-9 and AEG-1 are significant higher in alcohol victims, and expressions of MMP-2 and MMP-9 and AEG-1 have positive correlation with the alcohol concentration. The alcohol has acute toxicity to brain cells.
Alcohol Drinking/adverse effects* ; Brain ; Cell Adhesion Molecules/metabolism* ; Craniocerebral Trauma/etiology* ; Death ; Ethanol/poisoning* ; Heart/drug effects* ; Humans ; Matrix Metalloproteinase 2/metabolism* ; Matrix Metalloproteinase 9/metabolism* ; Membrane Proteins ; RNA-Binding Proteins ; Subarachnoid Hemorrhage/complications*

This research was to study the regulation of intravenous administration of human umbilical cord blood mesenchymal stem cells (HUCBMSCs) on secretion of neural specific protein in rats after traumatic brain injury (TBI), and to explore its mechanisms promoting the recovery of neurological function. The TBI models of rats were established. We then injected HUCBMSCs, labelled by Brdu (5-bromo-2-deoxyuridine), into the TBI rats via the tail vein using modified Feeney free-falling method. The levels of neural biochemical indicators (serum S100β protein, NSE, LDH, CK) of rats were detected in shamed group, injury group and HUCBMSCs-transplanted group. And the morphological changes of brain tissue of rats in the three groups were observed by using HE staining under light microscope. During the whole experiment no immunosuppressant was used for the four groups. From the research, transplant-related death of the rats was not found in transplantation group. In the injury group, rises were found in contents of serum S100β protein, NSE, LDH, CK in the early stage after the rats were injured, which were much higher than those in shamed group at correspondent time point (P < 0.01). In HUCBMSCs-transplanted group, although these biochemistry indexes were found rising for a short period in the early stage, along with the time, these indexes were obviously lower than in those injury group (P < 0.05). Under light microscopy pathological changes of rats in HUCBMSCs-transplanted group were much slighter than those in injury group. It was well concluded that in the situation of no immuno-suppressants, the intravenous-injected HUCBMSCs could reduce the secretion of serum S100β protein, NSE, LDH, CK, promote the repair of tissue injury effectively, and promote the functional recovery of neurons.
Animals ; Biomarkers ; chemistry ; Brain ; pathology ; Brain Injuries ; therapy ; Cord Blood Stem Cell Transplantation ; Humans ; Mesenchymal Stem Cell Transplantation ; Neurons ; chemistry ; Rats

Objective To compare the efficacy of the laryngeal mask airway Classic (CLMA), Streamlined Liner of Pharyngeal Airway (SLIPA) and laryngeal mask airway ProSeal (PLMA) in patients undergoing laparoscopic surgery. Methods One hundred and twenty-three ASA Ⅰ or Ⅱ patients aged 20-64 yr undergoing elective laparoscopic surgery were randomly divided into 3 groups: CLMA group (group C, n =41 ), SLIPA group (group S, n =42) and PLMA group (group P, n =40). Laryngeal mask was inserted after induction of anesthesia with sufentanil 0.3 μg/kg, atracurium 0.5 mg/kg and propofol 2 mg/kg. Mll the patients were ventilated with intermittent positive-pressure ventilation. Anesthesia was maintained with contiunous infusion of propofol 6-8 mg· kg-1· h-1 and with supplementary administration of sufentsnil 0.1-0.2 μg/kg and atracurium 0.2 mg/kg as required. The degree of difficulty in inserting the laryngeal mask was evaluated. The rate of successful placement at first attempt, device placement time, recovery time and complications were recorded . Peak inspiratory pressure (PIP) and the number of patients with an airway sealing pressure less than PIP were recorded before pneumoperitoneum and when intra-abdominal pressure achieved 12 mm Hg. Results The placement was easier and the placement time and recovery time were significantly shorter in group S, and the placement time was significantly longer,the airway sealing pressure was significantly higher and the number of patients with an airway sealing pressure less than PIP was lower in group P than in group C ( P ＜ 0.05). The placement time and recovery time were significantly longer, the placement was harder, the airway sealing pressure was significantly higher, and the number of patients with an airway sealing pressure less than PIP was lower in group P than in group S ( P ＜ 0.05). PIP was significantiy higher when intra-abdominal pressure achieved 12 mm Hg than that before pneumoperitoneum in all the three groups ( P ＜ 0.05). There was no significant difference in the incidence of complications and rate of successful placement at first attempt among the three groups ( P ＞ 0.05). Conclusion All the three airway devices can provide adequate ventilation during operation, with fewer complications. SLIPA placement is easiest. PLMA can assure good airway sealing and is most suitable for patients undergoing laparoscopic surgery.

AIM To observe the effects of Qibao Meiran Oral Liquid (Polygoni multiflori Radix Praeparata,Angelicae sinensis Radix,Psoraleae Fructus,etc.) on learning and memory function,hippocampus tissue pathological morphology,SOD activity and carbonyl protein content in SAMP8 mice.METHODS Twenty-seven SAMP8 mice were randomly and equally divided into model control group,donepezil hydrochloride group and Qibao Meiran Oral Liquid group.Another nine SAMR1 mice were selected as normal control group.Mice were given successive intragastric administration for 60 days.On the 56th day,the passive avoidance test was adopted,and the learning and memory capacities were determined after 5 d;The pathological morphology was observed by HE staining;ELISA assay was used to detect the activity of SOD and the content of carbonyl protein in brain tissue.RESULTS Compared with the model control group,the escape latency of mice in the Qibao Meiran Oral Liquid group was significantly prolonged,and the number of errors decreased significantly (P ＜0.01);the pathological morphology of hippocampus tissue was significantly improved;SOD activity increased significantly,and carbonyl protein content decreased significantly (P ＜ 0.01).CONCLUSION Qibao Meiran Oral Liquid can not only improve the learning and memory function of SAMP8 mice,but also reduce the degree of hippocampus tissue degenerative disease.

In this study, we designed and synthesized 12 novel aloperine derivatives with different core structures. Among them, compound 3 with a ten-membered ring core was obtained through a special ring expansion reaction after γ-H Huffman elimination of quaternary ammonium salt, and the structure was verified by X-single crystal diffraction. Furthermore, their antiviral activity against human β-coronavirus HCoV-OC43 was evaluated by CCK-8 assay. Quaternary ammonium salt 2a and 3 had a good inhibitory effect against HCoV-OC43, and 2a had the highest anti-HCoV-OC43 activity with an EC₅₀ values of 3.77 μmol·L^-1 and a SI value of over 53.1. Schrӧdinger molecular docking results showed that both 2a and 3 might display their anti-HCoV-OC43 activity by directly acting on host TMPRSS2 and SR-B1. The results expanded the structural types of endocyclic aloperine and the function against coronavirus, and provided useful scientific data for the development of pharmaceutical applications of these compounds.

Objective To investigate the curative effect of brucellar spondylitis,so as to provide scientific proof for improving the curative level of the disease.Methods Epidemiological information was collected from 113 patients diagnosed as brucellar spondylitis,who were divided into 5 groups according to different drugs and drug combinations of doxycycline,gentamicin,sulfamethoxazole and rifampicin.Then the curative effect was investigated.Twenty-one patients who had greater paoas muscle abscess or Para vertebral abscess,intraspinal abscess of spinal canal,necrotic intervertebral disk and major osteolasia received the minimal invasive surgery and the focus removal surgery.Results The occurrence of the disease in female was much higher than that in male.Grazing and breeding beasts was the principal route of infection.Lumbars was mostly involved.they usually was infected in the adjacent 2 piece.L4 was the most common and seriuous one.The curative effect of doxycycline group was better than that without doxycycline(72.60% vs 35.00%,X2=15.14,P＜0.05).Doxycyeline+gentamicin+sulfamethoxazole was reeommended as the first choice.However,the curative effect did not increase despi~course of the treatment prolonged.The heMing rate and effective rate after 1 course was 52.21%(59/113)and 92.04%(104/113).that after 2 courses 58.41%(66/113)and 95.58%(108/113),that after 3 courses 59.29%(67/113)and 95.58%(108/113).The healing rate in different course did not present differences(P＞0.05).21 patients undergoing surgery were followed-up,12 patients were after 2 years and 9 patients were between 1-2 years.The healing rate was 95.24%(20/21),1 case was healed basically,the effective rate was 100%.None reoccurred.Conclusions There are characterized features in clinical epidemiology of the brucell spondylitis.Long term,adequate in dosage,combination and multi-approach use of antibiotics is the most reliable way to treat and prevent it from recurring.But fof the patients soitable for surgery.the minimal invasive surgery or the focus removal could shorten the course of therapy,decrease the complications and increase the cure rate.

AIMTo investigate the molecular mechanisms of smooth muscle 22 alpha (SM22alpha) whereby cytoskeleton remodeling of vascular smooth muscle cells (VSMCs) is regulated.

METHODSSynthetic (dedifferentiated) VSMCs were converted to contractile (differentiated) VSMCs by serum deprivation. Cells were transfected with pEGFP-SM22alpha, and localization of SM22alpha and its relationship with F-actin were observed through fluorescence microscopy. Fractional extraction of proteins and Western blotting were used to detect polymerization of SM alpha-actin in antisense-pcD2-SM22alpha-transfected VSMCs.Furthermore, effect of SM22alpha on F-actin cross-linking was observed by F-actin polymerization experiment.

RESULTSFluorescence microscopy showed that SM22alpha co-localized with F-actin in contractile VSMCs. Western blotting of protein extracts from F-/G-actin fractions revealed that polymerization of SM alpha-actin was lower in antisense-pcD2-SM22alpha-transfected VSMCs, in which SM alpha-actin mostly existed as soluble G-actin. Moreover, F-actin polymerization in vitro also showed that GST-SM22alpha could promote cross-linking of F-actin to form thick and bundled stress fibres,while extracts from VSMCs transfected with antisense-pcD2-SM22alpha could not effectively induce the polymerization of F-actin.

CONCLUSIONSM22alpha acts as a modulator to participate in VSMC cytoskeleton remodeling. It can not only induce polymerization of G-actin to F-actin, but also promote cross-linking of F-actin to bundled stress fibres, indicating its vital role in cytoskeleton remodeling.

Actins ; metabolism ; Animals ; Cytoskeleton ; metabolism ; Male ; Microfilament Proteins ; metabolism ; Muscle Proteins ; metabolism ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley

AIMTo investigate the interaction between C-terminal domains of SM22alpha and cytoskeleton F-actin.

METHODSProkaryotic expression vector containing SM22alpha cDNA and GST sequence was constructed. The induction conditions were optimized to increase the product of soluble GST-SM22alpha fusion protein in E coli. Expression products were purified and rabbit anti-GST-SM22alpha polyclonal antibody was produced by the purified fusion protein. In order to explore the effect of SM22alpha on cytoskeleton reorganization, VSMCs were treated with serum withdrawal and then serum stimulation to induce contractile/synthetic phenotypic modulation. SM22alpha protein distribution in F-actin/G-actin fractions was detected by Western blotting. The interaction between SM22alpha and actin was examined by GST pull down assay and coimmunoprecipitation. Colocalization of endogenous SM22alpha with F-actin was observed by immunofluorescence.

RESULTSThe results showed that the expression of soluble GST-SM22alpha protein was the highest under condition induced by 30 degrees C, 0.5 mmol/L IPTG for 6 h. Immunofluorescence and Western blotting of protein extracts from F-actin/G-actin fractions revealed that SM22alpha colocalized with F-actin during VSMC redifferentiation. GST pull down assay and coimmunoprecipitation showed that SM22alpha interacted with F-actin by C-terminal domains to participate in cytoskeleton reorganization.

CONCLUSIONThe recombinant SM22alpha C-terminal domains have the ability to bind F-actin, by which SM22alpha interacts with actin and participates in cytoskeleton reorganization.

Actins ; metabolism ; Amino Acid Sequence ; Animals ; Cell Line ; Cytoskeleton ; metabolism ; physiology ; Microfilament Proteins ; genetics ; Molecular Sequence Data ; Muscle Proteins ; genetics ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; Protein Binding ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; genetics

Objective To construct the transformed mouse BaF3-P210 cell line stably expressing BCR/ABL and to initially investigate the influence of BCR/ABL on the cell biological characteristics of BaF3 cell line. Methods The retroviral vector with bcr/abl gene was transfected into the packaging cell line. The BaF3 cells were infected with the collected viral supernatant. The transgenic BaF3-P210 cell line stably expressing BCR/ABL were screened and subcloned. The integration of the bcr/abl gene in the genome of the target cell was determined by PCR and DNA sequencing,trypan blue staining assay,flow cytometry and MTT assay. Results The bcr/abl gene was integrated into the BaF3 cell genome; RT-PCR and Western blot verified the stable expression of the bcr/abl gene and protein in the screened subclone cell line BaF3-P210. Compared with the control group,the cell proliferation rate was promoted (P

Objective To evaluate the clinical efficacy of Qingrehuashi herbal formula combined with high intensity focused ul-trasound(HIFU ) in the treatment of advanced pancreatic cancer .Methods 86 patients with pancreatic cancer (22 case of III stage and 64 case of IV stage) were included in this study .18 cases were performed the HIFU therapy for 2-3 times .Other 68 cases re-ceived once HIFU therapy ,among 53 cases of liver metastasis ,8 cases were simultaneously conducted HIFU ablation therapy on liv-er metastasis .The patients were given Chinese medicines dominated by Qingrehuashi before and after HIFU therapy and during fol-low up period .Results The single evaluation on HIFU irradiation cases after 1 month:complete remision(CR) in 0 case ,partial re-mission(PR) in 8 cases(9 .3% ) ,stable disease(SD in 64 cases(74 .4% ) and progress disease(PD) in 14 cases(16 .3% ) .The median survival rate of 1 year and half a year was 52 .0% and 11 .4% .Among 73 cases of increased CA199 before treatment ,CA199 after treatment was decreased in 12 cases .Among 36 cases of increased CA242 before treatment ,CA242 after treatment was decreased in 15 cases .The effective rate of analgesic relief in all the cases was 70 .9% (62/86) .Conclusion The integrated therapy of Qingre-huashi herbal formula and HIFU is an effective method for treating advanced pancreatic cancer .