To set up a new method of detecting bacterial number in situ,NADH fluorescence method based on the fluorescent characteristic of NADH was used.When the concentration of NADH ranged from 10 nmol/L to 0.2 mmol/L,its concentration had a good line relationship to the fluorescence intensity(R2= 0.9905).Separating bacterial cells by centrifugation and extracting NADH with hot Tris-HCl buffer,the re-sult of bacterial count detected with NADH standard plot was 1?104 CFU/mL in an hour.In summary,NADH fluorescence method is rapid,sensitive,simple and reliable to detect total bacterial number.There-fore,the method can be widely applied in the field of food sanitation and safety,environment detection and so on.

Animals ; Blood Glucose ; Diet, High-Fat ; Ghrelin ; blood ; Lipids ; blood ; Physical Conditioning, Animal ; Rats

OBJECTIVETo study the effect of melatonin on the gastrointestinal motility and plasma levels of the stress hormone in overtraining rats.

METHODThirty adult SD rats were randomly divided into three groups (n = 10): control group, over-training group, melatonin intervention group. 30 min before each training, rats in the control and over-training groups were fed with normal saline (15 mg/kg) once a day and 5 times per week, while rats in the melatonin intervention group were administrated with melatonin, perfusion in the intervention group (15 mg/kg). Excessive training group and melatonin intervention group rats were subjected to excessive training at 5 times a week for 6 weeks. After 6 weeks, the gastric emptying rate, small intestinal propulsion ratio and levels of plasma motilin (MTL) and calcitonin gene-related peptide (CGRP), cortisol (CORT) and catecholamines (CA) were observed in all groups.

RESULTSCompared with the control group, the gastric emptying rate, small intestinal propulsion ratio and levels of plasma MTL, CORT and CA were increased significantly (P < 0.01) while the content of CGRP was reduced (P < 0.01) in over-training group. After treated with melatonin, this trend was reversed, that was, the gastric emptying rate, small intestinal propulsion ratio and levels of plasma MTL, CORT and CA were surpressed significantly (P < 0.01) while the content of CGRP was improved obviously (P < 0.01) in over-training group.

CONCLUSIONMelatonin plays an important role in protecting gastrointestinal tract from dysfunction, in which MTL, CGRP, CORT and CA are all involved.

Animals ; Calcitonin Gene-Related Peptide ; blood ; Catecholamines ; blood ; Fatigue ; Gastrointestinal Motility ; Hydrocortisone ; blood ; Melatonin ; pharmacology ; Motilin ; blood ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley ; Stress, Physiological

Objective To quantitatively detect serum Golgi protein 73 (GP73) and alpha-fetoprotein (AFP) in the patients with hepatocellular carcinoma (HCC) ,chronic hepatitis liver cirrhosis and normal healthy individuals ,to compare the detection rate of GP73 and AFP and to investigate the clinical significance of GP73 for diagnosing HCC .Methods The serum samples in normal healthy individuals and the patients with chronic hepatitis liver cirrhosis and HCC were collected and detected GP 73 and AFP by the enzyme-linked immunosorbent assay (ELISA) and the chemiluminescence method .Results The detection rates of GP73 and AFP in HCC were 75 .61% and 40 .24% respectively ,GP73 was significantly higher than AFP (P<0 .01);In chronic hepatitis liver cirrhosis ,the detection rates of GP73 and AFP were 12 .58% and 41 .67% respectively ,the detection rate of GP73 was significantly lower than that of AFP (P<0 .01);In normal healthy individuals ,the detection rates of GP73 and AFP were 1 .67% and 6 .67% re-spectively ,the difference between them had no statistical significance (P>0 .05) .The specificity of GP73 and AFP in the normal healthy individuals and the patients with chronic hepatitis kive cirrhosis were 93 .42% and 77 .78% respectively .Areas under the ROC curves of GP73 and AFP were 0 .934 1 and 0 .806 6 respectively .The HCC detection rate of the combined detection of GP73 and AFPT was 87 .80% .Conclusion The GP73 detection rate and content are extremely low in the normal healthy individuals ,the GP73 detection rate is 12 .58% in chronic hepatitis liver cirrhosis and up to 75 .6% in HCC .The ROC curve analysis and the detec-tion rate are basically consistent .GP73 has the high detection rate and strong specificity in the patients with HCC ,which is the pre-ferred marker for early diagnosis of HCC .The joint detection of GP73 and AFP can improve the early diagnosis rate of HCC .

Objective To study the expressions of neuroglobin gene in cellular tissues of human body.Methods Total 13 species tissue RNA, superscription RNA 2 ?g of every tissue was reversed into transcription cDNA and 1 ?l of cDNA was made into PCR template. Total RNA 2 ?g from every specimen equivalently were reversed into transcription cDNA. PCR products experienced agarose gel electrophoresis and electrophoresis and extraviolet-gelatum Image J was quantitatively analyzed. All data were indicated with ?s and treated with Oneway mono agent analysis of variance of stata statistical package,P

Background Proliferative vitreoretinopathy (PVR) is one of the major causes of retinal detachment surgery failure.Based on proteomic studies of PVR vitreous,the insulin-like growth factor binding protein-6 (IGFBP-6) protein was specifically expressed in the vitreous and serum of PVR patients.Furthermore,its expression level is higher in the vitreous and serum in severe PVR patients than that in mild PVR patients.Objective This experiment was to detect the expression of IGFBP-6 in a PVR rat model.Methods Seventy 7-week old male SPF Wistar rats were included and were randomized into the PVR model group and control group.A mixture of RPE-J cell suspension(5 μl) and platelet-rich plasma (5 μl) was intravitreally injected in the left eyes of adult Wistar rats to establish the PVR model,and normal saline solution was administered in the same way in the control group.The rat eyes were clinically examined 1 week,2,3 and 4 weeks after injection,and PVR was graded based on the criteria of Francine.The animals were sacrificed after 1 week,2,4 or 8 weeks for the preparation of retinal sections and liver extraction.Expression levels of IGFBP-6 mRNA in the rat retina and liver were assayed by real-time Q-PCR.The expression of IGFBP-6 protein in the rat serum and vitreous was detected by ELISA.The use of animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results Purified IGFBP-6 RNA was extracted from the liver and retina of Wistar rat and quantified by real-time Q-PCR.The expression level of IGFBP-6 mRNA in retina was (3.79± 1.33) × 10-4 in the PVR model rats,showing a significant decline in comparison with the control rats with a level of(8.32±2.96) × 10 4,4 weeks after injection (t =3.42,P＜0.01).The expression of IGFBP-6 mRNA in the 4th week was significantly lower than that of 1 week,2 or 8 weeks after the establishment of the PVR model(P＜0.05).No significant difference was found in the IGFBP-6 mRNA level in the liver between the PVR group and control group(27.60± 14.01 × 10 4 vs.25.01 ± 12.04 ×10-4,respectively),as well as among the different time points(P＞0.05).IGFBP-6 mRNA content in the retina was significantly reduced in grades 1,2 or 3 of the PVR groups compared with the control group(P＞0.05),but there was no significant difference among the different grades of PVR groups (P＞0.05).Concentrations of IGFBP-6 protein in grades 1,2 and 3 of the PVR model group were (221.00 ± 19.32),(229.63 ± 18.89) and (225.70 ± 26.71) μg/L,with a significant elevation in comparison with (173.25 ±21.11) μg/L of the control group (t =2.14,P＜0.05).However,there was no significant change among the different grades of PVR groups(t=1.24,1.46,P＞0.05).The concentrations of IGFBP-6 protein in the vitreous and serum were higher in PVR rat samples (vitreous:225.44±19.36 μg/L;serum:108.48 ± 15.78 μg/L) than in control rats (vitreous:173.25 ± 21.11 μg/L,serum:95.96 ±17.40 μg/L)(P＜0.05).Conclusions The concentrations of IGFBP-6 protein in the vitreous and serum increase in PVR rats.The results indicate that the increased IGFBP-6 in the vitreous might be a localized autocrine secretion of the eye.

ObjectiveTo study the nursing characteristics in surrounding period of operation for treating carotid artery stenosis with stenting angioplasty. MethodsThe nursing procedure for 78 cases with carotid artery stenosis were reviewed to form the duties and items of nursing, including cooperation between doctors and nurses, mentality nursing, instruction of convalescence in the pre-operation, during operation and post-operation. ResultsAll the cases has successfully finished their operative procedure and none serious complication has been found. ConclusionThe nursing play a important role in the stenting angioplasty of carotid artery.

OBJECTIVE:To analyze the rules and characteristics of adverse drug reactions(ADR)induced by Shuxuetong in-jection,and provide reference for clinical rational drug use. METHODS:ADR cases induced by Shuxuetong injection in Beijing collected by national ADR monitoring center in 2012 were retrieved,and 35 cases of the ADR were statistically analyzed. RE-SULTS:ADR induced by Shuxuetong injection had relatively high incidence in elder over 60 years old(68.57%);ADR might oc-curred after 5 min with off-label use and it mainly occured after 10 min-2 h(40.00%);clinical manifestations were mainly systemic damage and nervous system damage,and all patients were returned to normal after symptomatic treatment except one with unclear outcome. CONCLUSIONS:ADR induced by Shuxuetong injection is related to age and dosage. Clinical medication should be strict-ly in accordance with the instructions,strengthen medication monitoring and reduce incidence of ADR to ensure medication safety.

Objective To investigate the effects of 5-azacytidine on radiation-induced epithelialmesenchymal transition in rat alveolar type Ⅱ epithelial cell line (RLE-6TN) and explore their working mechanisms,and to provide experimental evidence for the potential drug-based treatment of radiationinduced pulmonary fibrosis.Methods RLE-6TN cells were cultured in vitro and divided into four groups according to the experimental purposes:control group (C),radiation group (R),5-azacytidine group (A),and radiation followed by 5-azacytidine group (R + A).The microstructural changes in cells were determined by transmission electron microscopy.Inverted phase-contrast microscopy revealed the morphological changes in cells.The mRNA expression levels of E-cadherin and α-SMA were measured by quantitative real-time polymerase chain reaction (qRT-PCR).The protein expression levels of E-cadherin,GSK3 β,and p-GSK3 β (Ser9) were measured by Western blot.The one-way analysis of variance was used for pairwise comparison.Results The cells in group R became spindle-like.Similar morphological changes were not observed in cells in group R + A.Osmiophilic lamellar bodies disappeared at last in cells in group R.RT-PCR results showed that compared with group C,group R had a significantly lower mRNA expression level of E-cadherin ((0.23 ± 0.06) vs.(1.00 ± 0.00),P =0.002)) and a significantly higher mRNA expression level of α-SMA ((2.91 ± 0.01) vs.(1.00 ± 0.00),P =0.000)).However,compared with group R,group R + A had a significantly higher mRNA expression level of E-cadherin ((0.47 ± 0.05) vs.(1.00 ± 0.00),P =0.024)) but a significantly lower mRNA expression level of α-SMA ((2.50 ± 0.02) vs.(1.00 ±0.00),P =0.037)).The results of Western blot showed that the protein expression level of Ecadherin was significantly reduced ((0.07 ± 0.01) vs.(0.48 ± 0.02),P =0.028)),while the protein expression level of p-GSK3β was significantly increased in Group R than in Group C ((0.85 ± 0.04) vs.(0.23 ± 0.03),P =0.031)).However,compared with group R,group R + A had a significantly lower protein expression level of E-cadherin ((0.25 ± 0.00) vs.(0.07 ± 0.01),P =0.024)) and significantly less up-regulation of the protein expression level of p-GSK3β ((0.39 ± 0.03) vs.(0.85 ± 0.04),P =0.014)).Conclusions X-ray radiation can induce the epithelial-mesenchymal transition in epithelial cells.5-azacytidine suppresses radiation-induced epithelial-mesenchymal transition by inhibition of the activity of p-GSK3β in RLE-6TN cells.

64 patients with severe chronic periodontitis were divided into 2 groups(n =32)randomly.Intramuscular injection of ketorolac tromethamine(KT group)and local block injection of lidocaine(L group)were respectively applied for periodontal subgingival scaling and root planning.The analgesic effect was assessed using VAS,the adverse drug reactions were recorded during and after operation.During op-eration the VAS between the 2 groups was not statistically different(P >0.05).6 and 1 2 h after operation the VAS of KT group was lower than that of L group(P <0.05).During operation minor intraoperative local adverse reactions were observed in all cases in L group,but not any in KT group(P <0.05);after operation minor adverse reactions were observed in 1 case of KT group and in 2 cases of L group(P >0. 05).