Background Neurotransmitter secretion disorder induced by chronic manganese poisoning has always been one of the important causes of body injury, but the mechanism of neurotransmitter secretion disorder caused by manganese is not clear at present. Objective To investigate the effects of presynaptic membrane intracellular protein 13-1 (Munc13-1) and synapse fusion protein binding protein 18-1 (Munc18-1) on dopamine secretion dysfunction induced by manganese chloride (MnCl₂) in human neuroblastoma (SH-SY5Y) cells. Methods A SH-SY5Y cell model induced by MnCl₂ was established. Cell viability was measured by MTT assay. Four experimental groups were set up: control group and low-, medium-, and high-dose manganese groups (0, 100, 200, and 400 μmol·L⁻¹ MnCl₂). They were treated with corresponding doses of MnCl₂ for 24 h. The secretion of dopamine was measured by enzyme-linked immunosorbent assay. The mRNA expression of Syntaxin-1 was detected by real-time quantitaive PCR. Total cell proteins were extracted, and the protein expression levels of Munc13-1, Munc18-1, and Syntaxin-1 were detected by Western blotting. The correlations of MnCl₂ exposure and dopamine secretion with the protein expressions of Munc13-1 and Munc18-1 were also analyzed by Pearson correlation. Results Compared with the control group, the cell viability rate decreased gradually with the increase of manganese exposure concentration, and the difference between the medium- and the high-dose manganese groups was statistically significant (P<0.05). The concentration of dopamine in cell culture medium of all manganese exposure groups decreased with the increase of manganese concentration, and compared with the control group and the low-dose manganese group, the medium- and the high-dose manganese groups were statistically significant (P<0.05). The expression of Syntaxin-1 at mRNA or protein level did not change significantly among groups (P>0.05). Compared with the control group, the protein expression of Munc13-1 decreased and that of Munc18-1 increased with the increase of manganese concentration (P<0.05). Compared with the low-dose manganese group, the changes of Munc13-1 protein in the high-dose manganese group and Munc18-1 protein in the medium- and high-dose manganese groups had statistical significance (P<0.05). Compared with the medium-dose manganese group, the protein changes of Munc18-1 in the high-dose manganese group were statistically significant (P<0.05). The correlation analysis showed that MnCl₂ dose was negatively correlated with Munc13-1 protein expression (r=−0.898, P<0.05), and positively correlated with Munc18-1 protein expression (r=0.678, P<0.05). Dopamine secretion was positively correlated with Munc13-1 protein expression (r=0.932, P<0.05), and negatively correlated with Munc18-1 protein expression (r=−0.817, P<0.05). Conclusion The inhibition of dopamine secretion in SH-SY5Y cells induced by manganese exposure is related to up-regulation of Munc18-1 and down-regulation of Munc13-1 expression levels, which may be one of the reasons for nerve injury caused by manganese.

Objective:To screen and clone a carrier molecule for the expression of small bioactive peptides at high levels. Methods: A carrier molecule, PaP3.30, was screened out from the genome of Pseudomonas aeruginosa phage PaP3 and its gene was cloned by PCR method and inserted into pQE 32 expression plasmid, this recombinant plasmid was named pQE PaP30. The peptide antibiotics hPAB ? gene was then inserted into pQE PaP30 and induced to express the fusion protein in Escherichia coli . The ability of PaP3.30 to express other bioactive peptides was evaluated by fusing 6 different origins, varies in sizes and isoelectric points selected peptides to it. Results: After fused to PaP3.30, the peptide antibiotics hPAB ? could express as fusion protein above 30% of total bacterial proteins. Six selected peptides were also expressed by the level of 35%～44% total bacterial proteins when fused to carrier molecule, PaP3.30. Conclusion: The new carrier molecular, PaP3.30, is versatile in the expression of small bioactive peptides.

Objective To investigate the developmental characters of neural stem cells(NSCs) in occipital of cortex from human fetal brain at different age.Methods Ninety cases of embryoes at gestational age 16-32 weeks and by induction of labor with water bag were collected for determining distribution,shapes,growth modes and the number of NSCs in the occipital of cortex with immunohisto- chemical method under light microscope.Results It was noted that NSCs existed in the occipital of cortex from human fetal brain at different ages.NSCs mainly distributed in layers of cone cells and inner granule cells.NSCs existed in the occipital of cortex of different fetal age included middling round cells,NSCs had enations from 0 to 1.Nucli were larger than plasm.Each NSC had nucleoli from 2-4 and rarefaction chromatin.Most of NSCs distributed in three growth modes including crowd,cluster and clone,occasionally with a single growth mode among other nerve cells.There were no differences including distribution,shapes,growth modes and the number of NSCs in the occipital of cortex between groups,but,NSCs gradually decreased with increasing of age.Conclusion NSCs exists in the occipital of cortex from different gestational age,and the number of NSCs decreases with increasing of age.

Objective To investigate the effect of enteral nutrition tolerance assessment form on the early enteral nutrition tolerance in patients with gastric cancer after operation. Methods According to the admission sequence, 108 patients with gastric cancer were divided into the control group and the experiment group with 54 cases in each group. Enteral nutrition was used in the control group, while enteral nutrition tolerance assessment form was used to evaulate and care patients in experiment group. Finally, the two groups were compared in the anal exhaust time and the rate of complications. Result Anal exhaust in the experiment group was significantly earlier than that in the control group and the rate of complications was significantly smaller than that of the control group (P < 0.05). Conclusions The enteral nutrition tolerance assessment form for systemic evaluation and intervention is effective in improvement of the patients′tolerance to enteral nutrition in early postoperative enteral nutrition support to patients with gastric cancer. It can promote the recovery of patients.

Objective:To define impact of spontaneous TIMI-3 flow before angioplasty on outcomes of percutaneous coronary intervention strategy and the prognosis in patients with acute myocardial infarction (AMI ). Methods: The consecutive 301 patients enrolled in the ongoing register of emergent coronary angioplasty within 12 hours from symptoms who were diagnosed as having ST elevation AMI in our hospital from 2000 to 2006 were analyzed, they were followed up for one year and the clinical characteristics and survival rates were analysed. Results: Among the 301 patients enrolled in the ongoing register of emergent coronary angioplasty, spontaneous reperfusion (TIMI-3 flow) was present in 14.6% at initial angiography. Compared with patients without TIMI-3 flow, those with TIMI-3 flow before coronary intervention were less likely to present in new-onset heart failure(2.3% versus 16%, P=0.016), Patients with initial TIMI-3 flow had significantly lower 30-day mortality (0% versus 9.3%, P=0.035) , and cardiogenic shock (0%versus 8.6%, P=0.044) and had a shorter hospital stay (P=0.008). Cumulative 1-year mortality was 0% in patients with initial TIMI-3 flow, 11.3% with TIMI 0-2 flow (P=0.019). By COX regression analysis, postprocendural TIMI-3 flow was an independent determinant of survival (OR=0.285,P=0.004) , however,TIMI-3 flow before coronary intervention was not found as an independent determinant of survival significantly. The lenitive symptoms and current smoking were the independent determinants of TIMI-3 flow before coronary intervention (P=0.005, P=0.048, respectively).Conclusion: Patients undergoing primary percutaneous transluminal coronary intervention in whom TIMI-3 flow is present before angioplasty may present with greater clinical and angiographic evidence of myocardial salvage, be less likely to develop complications related to left ventricular failure, and improve early and late survival.

Humans ; Accidents, Traffic ; Colon, Sigmoid ; Forensic Medicine ; Autopsy ; Abdominal Injuries ; Thoracic Injuries ; Rupture

Objective To evaluate the clinical significance of combined measurement of antiperinuclear neutrophil cytoplasmic antibody(ANCA)and anti-saccharomyees cerevisia antibody(ASCA)for the diagnosis of inflammatory bowel disease(IBD)patients and difierentiation of Crohn's disease(CD)with ulcerative colitis(UC).Methods A total of 159 patients with IBD(97 UC,62 CD),167 patients with other non-IBD gastrointestinal conditions(NIBDC)and 25 healthy controls(HC)were recruited in our research.ASCA and ANCA were detected by enzyme-linked immunosorbent assay(ELISA)and indirect immunofluorescence assay.respectively.Results The prevalence of ASCA-IsA or IgG in CD group,UC group,NIBDC and HC were43.5%,14.4%,29.3%and 0,respectively.The prevalence of ASCA-IgA or IgG in CD group were higher than those in other groups(X2=16.76 or 4.12,P＜0.01 or＜0.05).The prevalence of ANCA in CD group.UC group,NIBDC and HC were 8.1%,56.7%,4.8%and 0,respectively.The prevalence of AMA in UC group were much higher than those in other groups(X2=38.08 or 90.47,P＜0.01).The sensitivity specificity and positive predictive value(PPV)in ASCA+/ANCA-were 40.3%.93.8% and 80.6%,respectively,and in ANCA+/ASCA-were 48.5%,98.4% and 97.9%,respectively.Condusions ASCA or ANCA testing alone are not sensitive enoulgh for diagnosing CD and UC,but their combination asses are specific for differential diagnosis between CD and UC.Combined testing of ASCA-IgA with IgG can improve the sensitivity in screening CD patients.The ASCA positive pattern in Chinese CD group are correlated with surgery.

Objective: To screen the best genetic engineering bacterium for the production of peptide antibiotic hPAB-? and evaluate its fermentation level in bottle. Methods:After analysis of the interest fusion protein expression levels of 8 recombinant bacteria containing 1-8 copies of human peptide antibiotic hPAB-? expressing plasmid respectively,2-5 copies expressing bacteria were chosen for the further study of their bacteria yield,expression forms of the target protein, dissolution of the inclusion bodies and the efficiency of fusion protein purification by affinity chromatography, then the best engineering bacterium with the certain copies of interest peptide expressing plasmid was screened out and its optimal fermentation parameters in bottle were also studied. Results:The recombinant bacterium transformed by 3 copies of interest peptide expressing plasmid was the best candidate for its bacteria yield (3.153 g/L) and fusion protein expression level (27.7%) were the highest among 1-8 copies candidates. The inclusion bodies of 3 copies target fusion protein could be easily dissolved by 8 mol/L urea and captured by Ni-NTA column. The elution of the fusion protein could be directly cleaved to monomer by adding 2 mol/L hydroxylamine, adjusting pH to 9.0 and incubating at 45℃ for 2 h. The optimal fermentation conditions of the selected recombinant bacteria were: culture the organisms with modified M9-CAA media at 37℃ and 160 r/min to (A 600 )≈2.5, then add IPTG to the final concentration 100 ?mol/L to induce the expression of target fusion protein for 5 h. Conclusion:The engineering bacterium containing 3 copies interest peptide recombinant expressing plasmid is the best candidate for the production of peptide antibiotic hPAB-?,and its fermentation parameters are confirmed.

Objective To probe into effective surgical methods to reduce post-operational recurrence rate of nodular goiter.Methods One hundred and fifty-seven patients with nodular goiter who were admitted and had operations in our hospitals between Jan,2007 and June,2011 were assigned to one of the following 4 groups:total thyroidectomy(TT),nearly total thyroidectomy (NTT),subtotal thyroidectomy (STT),removal of lesion only or partial thyroidectomy (also termed as local removal,LR).Postsurgical recurrences and complications were compared among these groups.Results The recurrences were statistically analysed.There was a highly significant difference between TF and STT、TT and LR、NTT and LR; no difference was observed between TT and NTT,STT and STT + E,STT and LR.A statistical analysis was also carried out on the postsurgical compications in these patients.highly significant differnce was found between TT and STT,TT and NTT,TT and LR.There was a significant diffrence between STT and NTT.There was no difference between STT and LR,NTT and LR.Conclusion TT,STT,NTT are all effective operations for nodular goiter and should be advocated.Satisfactory efficacy may be obtained with oral thyroxine or euthyrox for postsurgical lifetime.However,LR or partial thyroidectomy should be adopted cautiously.

Phytochemical investigation on the EtOH extract from the aerial part of Callicarpa kwangtungensis led to the isolation and characterization of 10 caffeoyl phenylethanoid glycosides, 2'-acetylacteoside (1), tubuloside E (2), acteoside (3), tubuloside B (4), isoacteoside (5), alyssonoside (6), 2'-acetylforsythoside B (7), brandioside (8), forsythoside B (9), and poliumoside (10). Compound 4 was isolated from the plants of Verbenaceae,and 6 was obtained from the Callicarpa genus, for the first time, while compounds 1, 2, 5 and 7 were firstly reported from the plant.
Caffeic Acids ; chemistry ; isolation & purification ; Catechols ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; Ethanol ; chemistry ; Glucosides ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Phenols ; chemistry ; isolation & purification ; Verbenaceae ; chemistry