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 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.

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.

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

OBJECTIVE:To study the adverse drug reactions(ADRs)induced by weight-reducing drugs and to provide references for rational administration in clinics.METHODS:Classification statistic and analysis were conducted on462cases with weight-reducing drug-induced ADRs reported in62kinds of domestic and overseas medicinal periodicals published during the period of1980～2004.RESULTS:With pseudo-5-HTA appetite-suppressing drugs showing the high incidence of ADRs,which stood at36.36%of the total.Multi-systems were involved in ADRs,with the cardiovascular system showing the highest proportion(53.68%).The prognostic mortality accounted for18.61%of the total.CONCLUSION:ADRs induced by weight-reducing drugs were associated with many factors,to avoid the occurrences of which,medical workers should give more directions and monitoring on administration of drugs.

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).