Latrodectus tredecimguttatus (commonly known as black widow spiders) have toxins not only in their venom glands, but also in other parts of their body, in their eggs and even in the newborn spiderlings. The study on the toxins in venom and materials outside the venom glands of the spiders to elucidate their differences and similarities, evolutional relationship and biological functions is of important theoretical and applicable significance. The development of modern protein chemistry and proteomics techniques has provided efficient means for the study of protein and peptide toxins of L. tredecimguttatus. By using such techniques, the molecular base and action mechanism of the toxins can be revealed at the levels of both single purified proteins and omics. Up to now, although protein chemistry and proteomics study on L. tredecimguttatus toxins have achieved a certain progress, the relevant work particularly that on the toxins in the materials outside the venom glands has to be further deepened.
Animals ; Arthropod Proteins ; chemistry ; Black Widow Spider ; chemistry ; Proteomics ; Venoms ; chemistry

OBJECTIVE:To explore the protective effect of L-ornithine-L-aspartate on ischemia-reperfusion injury after partial hepatectomy. METHODS:104 patients underwent partial hepatectomy(vessel occlusion in portal fissure)were randomly divided in-to control group(53 cases)and trial group(51 cases). Control group was given routine liver-protective drugs,and trial group was additionally given L-ornithine-L-aspartate 10 g added into 5%Glucose injection 250 ml intravenously before surgery,for 7 consecu-tive days. The fasting peripheral venous blood was collected in 2 groups on first day before surgery and first,forth and sixth day af-ter surgery to detect liver function;the changes of main aminopherase index(ALT and AST)were compared between 2 groups at different portal fissure vessel occlusion time after surgery. RESULTS:There was no statistical significance in the difference of liver resection range,blood loss,first porta hepatis vessel occlusion time and anesthesia time in both groups during operation (P>0.05). Compared to before surgery,liver function indexes raised to different extent in 2 groups after surgery,with statistical signifi-cance(P<0.05). The levels of AST and ALT in trial group were significantly lower than in control group on first,forth and sixth day after surgery,while albumin level was significantly higher than control group,with statistical significance(P<0.05). Total bili-rubin levels of both groups were approximate basically,there was no statistical significance(P>0.05). Among those patients under-went porta hepatis vessel occlusion with Pringle method and with occlude occlusion time ≥15 min,the AST and ALT level of con-trol group was higher than those of trial group after surgery,but albumin level was below trial group,with statistical significance (P<0.05). CONCLUSIONS:L-ornithine-L-aspartate could improve liver function fast and effectively for partial hepatectomy pa-tients,especially for the patients underwent porta hepatic vessel occlusion with Pringle method for a long time(obvious ischemia-re-perfusion injury).

Objective:To study the relationship between atrophy of the thymus and disease severity in EAE.Methods:MOG35-55 peptide induced EAE in C57BL/6 mice and analyzed the relationship between the severity of EAE and thymic atrophy,Flow cytometry analysis was used to evaluate thymic CD4+CD8+DP cells,CD4+CD8-,CD4-CD8+SP cells in relation to the severity of the disease.Results:The number of thymocytes in mice with decreased tail tone was (20.25 ±3.49) ×106 ,hindlimb weakness(4.93 ± 0.85)×106,complete hindlimb paralysis(1.8 ±0.19) ×106,and forelimb and hindlimb paralysis(0.52 ±0.07) ×106,there were statistically significant differences between groups ( P<0.05 ).As the disease progresses, CD4+CD8+DP cells ratio decreased, CD4+CD8-,CD4-CD8+SP cell ratio increased,different disease groups was statistically significant difference (P<0.05).Conclusion: The atrophy of thymus was closely related to the severity of EAE.Migration of activated T cells in EAE may cause atrophy of thymus.

Dorsal root ganglion (DRG) neurons are primary sensory neurons that conduct neuronal impulses related to pain, touch and temperature senses. To comprehensively identify proteins of plasma membrane (PM) from small amount of dorsal root ganglion (DRG) neurons, a proteomics strategy that utilizes aqueous polymer two-phase partition in combination with differential velocity centrifugation was adopted to enrich the PM, followed by SDS-PAGE, CapLC-MS/MS and bioinformatics analysis. Western blot analysis showed that the concentration of PM in purified plasma membrane(PPM) was 2.3 times higher than that in crude plasma membrane(CPM), 15 times higher than that in whole tissue lysate (WTL). By searching against the rat IPI protein sequence database, a total of 729 non-redundant proteins were identified from the PM preparation, of which 547 had a gene ontology (GO) annotation indicating a cellular component, and 159 (21.8%) were unambiguously identified as PM proteins. A data set of plasma membrane proteins of DRG as well as a tool to study PM proteins were provided in a small amounts of sample.

The low abundance and highly hydrophobic nature of most membrane proteins make their analysis more difficult than that for common soluble proteins. Successful membrane protein identification is largely dependent on the sample preparation including the enrichment and dissolution of the membrane proteins. A series of conventional and newly developed methods has been applied to the enrichment of low-abundance membrane proteins at membrane and/or protein levels and to the dissolution of hydrophobic membrane proteins. However, all the existing methods have inherent advantages and limitations. Up to now, there has been no unique method that can universally be employed to solve all the problems and more efforts are needed in improving sample preparation for the analysis of membrane proteomes.
Chromatography, High Pressure Liquid ; Mass Spectrometry ; Membrane Proteins ; analysis ; metabolism ; Membranes, Artificial ; Oxidation-Reduction ; Polyvinyls ; chemistry ; Proteome ; analysis ; Proteomics

Jingzhaotoxin-V(JZTX-V) isolated from the venom of the spider Chilobrachys jingzhao is a novel potent inhibitor that acts on tetrodotoxin-resistant and tetrodotoxin-sensitive sodium channels in adult rat dorsal root ganglion(DRG) neurons. It is a 29-residue polypeptide toxin including three disulfide bridges. To investigate the structure-function relationship of the toxin, a mutant of JZTX-V in which Arg20 was substituted by Ala, was synthesized by solid-phase chemistry method with Fmoc-protected amino acids on the PS3 automated peptide synthesizer. The synthetic linear peptide was then purified by reversed-phase high performance liquid chromatography and oxidatively refolded under the optimal conditions. The refolded product was analyzed by matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry(MALDI-TOF MS) and electrophysiological experiments for its relative molecular weight and prohibitive activity of sodium channels respectively. The present findings show that the prohibitive effect of R20A-JZTX-V on TTX-S sodium channels in DRG neurons is almost the same as that of native JZTX-V, suggesting that Arg20 does not play any important role in inhibiting TTX-S sodium currents in DRG neurons. In contrast, the prohibitive level of R20A-JZTX-V on TTX-R sodium channels is reduced by at last 18.3 times, indicating that Arg20 is a key amino acid residue relative to the bioactivity of JZTX-V. It is presumed that the decrease in activity of R20A-JZTX-V is due to the changes of the property in the binding site in TTX-R sodium channels.
Amino Acid Substitution ; Animals ; Arginine ; genetics ; Ganglia, Spinal ; drug effects ; Mutagenesis, Site-Directed ; Mutant Proteins ; pharmacology ; Neurons ; drug effects ; Patch-Clamp Techniques ; Peptides ; chemistry ; genetics ; pharmacology ; Rats ; Sodium Channel Blockers ; pharmacology ; Sodium Channels ; drug effects ; Spider Venoms ; chemistry ; genetics ; isolation & purification ; pharmacology ; Spiders ; Tetrodotoxin ; pharmacology

OBJECTIVETo investigate whether patients, who are at risk of major acute coronary events, are safe to undergo and benefit from early intervention after using simvastatin.

METHODSThe study was a randomized, open, two-dosage-controlled trial to evaluate the safety and benefits of simvastatin administered to 197 patients (10 mg group, n = 98 and 20 mg group, n = 99), within 48 hours of hospitalization for a diagnosis of unstable angina or acute myocardial infarction (MI), with total cholesterol (TC) >/= 180 mg/dL or low-density lipoprotein cholesterol (LDL-C) >/= 100 mg/dL. Lipid levels were measured immediately, followed by the 3rd, 6th and 12th month after admission and all adverse events were recorded during follow-up.

RESULTSTC levels fell by 10.15% and 14.52% in the 10 mg and 20 mg groups (P < 0.05), and LDL-C levels fell 13.87% and 19.38% in the 10 mg and 20 mg groups, respectively (P < 0.01), 12 months after using simvastatin. The rates of achieving target TC reached 26.3% and 36.5% in the 10 mg and 20 mg groups (P < 0.01), and that of LDL-C reached 28.2% and 40.3% in the 10 mg and 20 mg groups, respectively (P < 0.01). There were higher rates of MI and re-hospitalization resulting from angina pectoris and revascularization in the 10 mg group compared with the 20 mg group.

CONCLUSIONSThe results suggest that early intervention with the HMG-CoA reductase inhibitor, simvastatin, in acute coronary syndromes is possible and safe. It also indicates that the clinical dosage of simvastatin are relatively smaller than that for satisfactory lipid control in patients with acute coronary syndromes.

Acute Disease ; Aged ; Cholesterol ; blood ; Cholesterol, LDL ; blood ; Coronary Disease ; blood ; drug therapy ; Female ; Follow-Up Studies ; Humans ; Hypolipidemic Agents ; therapeutic use ; Male ; Middle Aged ; Simvastatin ; therapeutic use

Objective:To investigate the relationship of mitochondrial DNA (mtDNA) copy number with clinicopathologic characteristics and its influence on the prognosis of hepatocellular carcinoma (HCC) patients.Methods:The retrospective case-control study was conducted. The clinicopathological data of 71 HCC patients undergoing surgical treatment in the Eastern Hepatobiliary Surgery Hospital of Naval Medical University from March to June 2011 were collected. There were 61 males and 10 females, aged from 26 to 80 years, with a median age of 55 years. The mtDNA copy number of tumor tissues and adjacent normal tissues were measured for all patients. Observation indicators: (1) the mtDNA copy number of tumor tissues and adjacent normal tissues and relationship between the mtDNA copy number and clinicopathological characteristics of HCC patients; (2) follow-up; (3) related factors for the prognosis of HCC patients. Follow-up using outpatient examination or telephone interview was conducted to detect postoperative survival of patients up to September 2019. Measurement data with normal distribution were described as Mean± SD, and comparison between groups was analyzed using independent samples t test or the matched samples t test. Measurement data with skewed distribution were described as M(range). Count data were represented as absolute numbers, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Univariate and multivariate analyses were conducted using the COX regressional model. Variables with P<0.10 in the univariate analysis were included for the multivariate analysis. Survival rates were calculated using the Kaplan-Meier method, and Log-rank test was used for survival analysis. Results:(1) The mtDNA copy number of tumor tissues and adjacent normal tissues and relationship between the mtDNA copy number and clinicopathological characteristics of HCC patients: of 71 HCC patients, the mtDNA copy number was 0.85±0.08 in tumor tissues, versus 1.16±0.08 in adjacent normal tissues, showing a significant difference between them ( t=2.96, P<0.05). Of 71 HCC patients, 48 cases were mtDNA-low and 23 cases were mtDNA-high. Cases with tumor capsule as integrity or not-integrity, cases with or without microvascular (MVI) in mtDNA-low and mtDNA-high patients were 20, 28, 21, 27 and 16, 7, 4, 19, respectively, showing significant differences ( χ2=4.84, 4.74, P<0.05). (2) Follow-up: 71 patients were followed up for 2.1 to 85.3 months, with a median follow-up time of 47.8 months. The 1-, 3-, 5-year overall survival rates of 71 HCC patients were 87.3%, 64.7, 37.4%, respectively. Moreover, the 1-, 3-, 5-year overall survival rates were 81.2%, 50.0%, 29.2% of the mtDNA-low patients, versus 95.7%, 86.5%, 54.7% of the mtDNA-high patients, showing a significant difference between the two groups ( χ2=5.86, P<0.05). (3) Related factors for the prognosis of HCC patients. Results of univariate analysis showed that the number of tumor, portal vein tumor thrombus, MVI, Barcelona Clinic Liver Cancer stage, mtDNA copy number were related factors for the prognosis of HCC patients ( hazard ratios=2.211, 2.911, 3.899, 3.587, 0.440, 95% confidence intervals as 1.024?4.777, 1.485?5.704, 2.115?7.186, 1.615?7.966, 0.223?0.871, P<0.05). Results of multivariate analysis showed that MVI and mtDNA copy number were independent influencing factors for the prognosis of HCC patients ( hazard ratios=2.754, 0.437, 95% confidence intervals as 1.374?5.521, 0.205?0.932, P<0.05). Conclusions:The mtDNA copy number of HCC patients is related with tumor capsule and MVI. The mtDNA copy number and MVI are independent influencing factors for the prognosis of HCC patients.