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

Objective To study the relationship between hepatitis B virus(HBV)pre-S gene mutations and progression of liver disease.Methods The entire pre-S1,pre-S2 genes were amplified hy nested polymerase chain reaction(PCR)and the products were digested by NlaⅢ.The method for detecting pre-S2 start codon mutation was established based on the digested restriction fragment length polymorphism(RFLP).Pre-S gene deletion was revealed by electrophoresis on polyacrylamide gel(PAGE).Pre-C A1896 and basic core promoter T1762/A1764 mutations were identified by direct sequencing of PCR products.The 138 sera from patients with HBV-related disease,including asymp- tomatic carriers(ASC),chronic hepatitis(CH),liver cirrhosis(LC),hepatocarcinoma(HCC),were tested by these methods.Results The detection rate of pre-S deletion mutation was higher in patients with HCC(56.3%)and LC(42.9%)than those with CH(11.8%)and ASC(8.1%,P

From the perspective of clinical doctors, in the teaching idea, national policy, school policies and different stages of clinicians cultivation , the article introduced the concept of translational medicine in the process of the clinical doctors tralning in the United States. It described US medical schools' (U.S. Virginia University School of medicine, for example) implementation ap-proach of translational medicine ideas in different stages, such as before entering school, during the period of school, practice exam and physician clinician tralning. It provided the reference for the de-velopment of translational medicine education in the process of China clinician education and tralning, including:medical students' integration into the related clinical research in preschool through volunteer service, the choice of multiple combination model of clinical science and basic research, interdisci-plinary examination of medical practitioners and the provisions of the research work in resident and specialist tralning stage.

Objective To evaluate the effects and mechanisms of bovine lactoferrin(bLF)on cell viability,proliferation,and the protective roles in intestinal epithelial cell-6(IEC-6)treated by lipopolysaccharide(LPS).Methods The rat jejunum epithelial cell lines IEC-6 were cultured in vitro.The effects of bLF on cell viability and proliferation in IEC-6 cells were detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide(MTT)assay and 5-Bromo-deoxyuridine(Brdu)assay,respectively.Inflammatory cytokines and their mRNA of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and interleukin-8(IL-8)were analyzed by real-time PCR and enzyme-linked immunosorbent assay(ELISA).Western blot was used to measure the levels of mitogen-activated protein kinase(MAPK)activation and nuclear factor kappa β(NF-κB)nuclear translocation.Results Dose dependent effects of bLF on cell viability and proliferation were observed in IEC-6 cells in vitro(F=3.825,5.861,all P<0.05),especially in a dose of 100 mg/L,and bLF significantly stimulated cell viability and proliferation compared with non-treatment group(q=5.240,3.765,all P<0.05).The mRNA levels of IL-1β,IL-6 and TNF-α were decreased by co-stimulation of bLF and LPS compared with the LPS treatments alone in IEC-6 cells in vitro(q=14.28,10.12,16.45,all P<0.001).The secretion of IL-6 and TNF-α was also decreased by co-stimulation of bLF and LPS(q=15.06,6.74,all P<0.01).In vitro,bLF treatment at dose of 100 mg/L could inhibit the activation of MAPK/NF-κB signal pathway induced by LPS(q=12.96,18.54,all P<0.001).Conclusion In vitro,bLF can promote IEC-6 viability and proliferation,and have anti-inflammatory effects via inhibited activation of MAPK/NF-κB nuclear translocation.

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.

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.