Terminal restriction fragment length polymorphism(T-RFLP) is a resent molecular approach that can assess subtle genetic differences between strains as well as provide insight into the structure and function of microbial communities. This method overcomes the confinement of conventional culture-dependent methods and has both high sensitivity and throughput making it ideal for comparative analyses. Though there is still no application in our country, more and more investigators are highlighting it. In this article, the fundamental principle of this technique is introduced. The recent application and the development of this technique are also summed up .

Objective To investigate dynamic changes of heme oxygenase-1 and carbon monoxide in acute liver injury induced by carbon tetrach loride(CCl_4)in rats.Method Male SD rats were randomly allocated to induce acute liver injury by CCl_4 injection.Hepatic HO activity was examined at different time point following CCl_4 treatment.Expression and location of HO-1 protein was determined by western blot and immunohistochemical methods.Serum ALT,AST levels and hepatic SOD,MDA concentrations were also analyzed.Results Administration of CCl_4 to rats caused a marked hepatic damage,characterized by significant elevation of serum ALT,AST levels and liver MDA content combined with a remarkable reduction in liver SOD activity.HO activity was elevated significanfly in a time-dependant manner after CCl_4 injection,while the expression of HO-1 protein increased remarkably from 6 to 36 hours.CO concentration in the liver homogenate of control rats remained very low but was elevated significantly after CCl_4 treatment,which was in accordance with changes of HO-1. Conclusions HO-1 activity and protein expression as well as CO production are higher in rats with acute liver injury induced by CCl_4 than in control group.HO-1/CO system plays an important role in the pathogenesis of acute hepatic damage and may have potent protective effect against liver injury.

OBJECTIVETo explore a new method of establishing HepG2 cell model of steatosis and observe the expression and significance of nuclear factor erythroid-2p45-related factor 2(Nrf2)/antioxidative response element (ARE) pathway related factors in HepG2 cells of steatosis.

METHODSHepG2 cells were induced with DMEM containing 25% fetal bovine serum, 0.1% MCT/LCT Fat Emulsion and 0.1 mmol/L free fatty acid (FFA) at different stages and the control group cells were cultured with normal DMEM medium. After the cell models were successfully established, lipid droplets in cytoplasm were observed with Oil Red 0 staining, and the triglyceride (TG) accumulation in HepG2 cells were tested by biochemical assay. Intracellular reactive oxygen species (ROS) concentration were detected by flow cytometry. Nitric oxide (NO), superoxide dismutase(SOD), malonyldialdehyde(MDA) and glutathione peroxidase(GSH-Px) were tested by biological reagent kit, while the protein expression of nuclear factor erythroid-2p45-related factor 2(Nrf2), heme oxygenase-1 (HO-1) and

NAD(P)Hquinone oxidoreductase-1(NQO1) were analyzed by Western blot.

RESULTSCompared with that in the control group, red cytoplasmic lipid droplets were visible in model group; TG,ROS, NO, MDA concentration (P < 0.05, P < 0.01) and the protein expression of Nrf2, HO-1 and NQO1 (P < 0.05, P < 0.01)were significantly higher in model group, while SOD, GSH-Px concentration reduced significantly (P < 0.01).

CONCLUSIONThe in vitro cell model of steatosis and oxidative stress was successfully established. The activation of Nrf2/ARE pathway related factors maybe relevant to the overreaction of oxidative stress in HepG2 cells of steatosis.

Antioxidant Response Elements ; Culture Media ; Fatty Acids, Nonesterified ; Fatty Liver ; metabolism ; GA-Binding Protein Transcription Factor ; Glutathione Peroxidase ; metabolism ; Heme Oxygenase-1 ; metabolism ; Hep G2 Cells ; Humans ; Malondialdehyde ; metabolism ; NAD(P)H Dehydrogenase (Quinone) ; metabolism ; NF-E2-Related Factor 2 ; metabolism ; Nitric Oxide ; metabolism ; Oxidative Stress ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Triglycerides ; metabolism

OBJECTIVETo investigate the startup detail of circulation dysfunction and its role in the progress of delayed neuropsychologic sequelae (DNS) after carbon monoxide (CO) poisoning with comparison with the model of ischemia-reperfusion.

METHODSThe ischemia-reperfusion rat model was established by Pulsinelli-Brierley method, and the CO poisoning rats model by i.p. injected with CO repeatedly respectively, and the rats were identified with DNS following the experiment of pathology and the ethnology.

RESULTSThe whole blood viscosity, plasma viscosity, hematocrit and fibrinogen increased significantly immediately after reperfusion, and recovered gradually with the ischemia-reperfusion rat model. The whole blood viscosity decreased significantly immediately after CO treated i.p. Especially at low shear rate, the hematocrit also declined remarkably in the early stage after CO treatment. But 1day later, these parameters turned to the trend of the ischemia-reperfusion rats. There was a prominent elevation of both indexes until the 14th day following CO injection i.p.

CONCLUSIONThere are significantly sustained hyper-coagulation and hyper-viscosity with circulation in rats after CO poisoning compared with ischemia-reperfusion model during the period of DNS, which might contribute to increase cerebral circulation resistance, blocked blood flow, and deteriorate hypoxemia in progression of DNS.

Animals ; Blood Circulation ; Carbon Monoxide Poisoning ; physiopathology ; Disease Models, Animal ; Hemorheology ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; physiopathology

OBJECTIVETo observe the therapeutic effect of local antibiotic injection into the female prostate on female urethral syndrome (FUS), and search for an effective treatment for this disease.

METHODSThis study included 163 FUS patients treated in the out-patient department between July 2009 and December 2010. According to the visiting order, the patients were randomly assigned to Groups A (n = 58), B (n = 55) and C (n = 50). All underwent routine treatment. Inaddition Group A received local injection of 2 ml of 80 000 U gentamycin + 2 ml of lidocaine, and Group B 2 ml of normal saline + 2 ml of lidocaine, both injected into the distal segment of the urethral back wall where the female prostate is located, twice a week for 3 weeks. The therapeutic effects were evaluated according to the changes of the patients' independent symptom scores at 2 and 4 weeks after the treatment. Disappearance of the symptoms was considered as "curative" , > 1/2 reduction in the symptom score as "obviously effective", 1/2 - > 1/4 reduction in the symptom score as "effective", and < 1/4 reduction or increase in the symptom score as "ineffective".

RESULTSAt 2 weeks after the treatment, the total effectiveness rate was significantly higher in Group A (77.5%) than in B (67.3%) and C (68.0%) (P < 0.05), but with no statistically significant difference between B and C (P > 0.05). At 4 weeks, the total effectiveness rate of Group A was slightly decreased, but still remarkably higher than that of group B or C (P < 0.05).

CONCLUSIONLocal injection of gentamycin into the female prostate is effective for the treatment of female urethral syndrome.

Administration, Topical ; Adult ; Aged ; Aged, 80 and over ; Female ; Gentamicins ; administration & dosage ; therapeutic use ; Humans ; Injections ; Middle Aged ; Prospective Studies ; Treatment Outcome ; Urethral Diseases ; drug therapy ; Young Adult

Objective: To study the clinical characteristics, methods of diagnosis and treatment of hyperornithinemia-hyperammonemia- homocitrullinuria (HHH) syndrome. Method: From July 2011 to August 2016, 3 Chinese patients with HHH syndrome were enrolled in this study. The clinical course, biochemical features, brain MRI findings, and gene mutations were analyzed. Result: The three patients′ age at onset of symptoms was 3 months to 7 years, and the age of diagonosis was 3 years and 10 months to 9 years and 10 months. All of them presented with intolerance to protein-rich foods from the infant period, development retardation and abnormal posture. Case 1 and 2 had moderate mental retardation. Serum ammonia 25-276 μmol/L (reference range<60 μmol/L), alanine aminotransferase (ALT) 20-139 IU/L (reference range 9-50 IU/L), ornithine 29.12-99.44 μmol/L(reference range 15-100 μmol/L), urinary orotic acid 1.49-29.75 mmol/mol Cr (reference range 0-7 mmol/mol Cr), uracil 6.09-103.97 mmol/mol Cr (reference range 0-1.5 mmol/mol Cr). The cranial MRI revealed lesions in the basal ganglia, abnormal white matter signal, progressive demyelination and cerebral atrophy. On their SLC25A15 gene, a novel homozygous missense mutation c. 416A>G (p.E139G) was identified in case 1, a known pathogenic homozygous nonsense mutation c. 535C>T was found in case 2 and 3. Liver transplantation had been performed when case 1 was 6 years old. Significant improvements were observed in dietary habit, mental and motor functions, and biochemical parameters. After the dietary intervention with the supplements of arginine, L-carnitine, case 2 was improved, spastic paraplegia of case 3 had no mitigation. Liver transplant was recommended. Conclusion HHH syndrome has an aversion to protein-rich food, and the patients have recurrent vomiting and progressive neurological dysfunction. Clinical diagnosis of HHH syndrome is difficult and patients may present with incomplete biochemical phenotype. The genetic analysis is key for the diagnosis. Depending on their condition, individuals with HHH syndrome can be treated with a low-protein diet, drugs and liver transplantation.

OBJECTIVETo develope a tree analysis pattern of mass spectral urine profiles to discriminate bladder transitional cell carcinoma (TCC) from non-cancer lesions using surface-enhanced laser desorption and ionization time-of-flight mass spectrometry (SELDI-TOF-MS) technology.

METHODSUrine samples from 61 bladder transitional cell carcinoma (TCCs) patients, 53 healthy volunteers and 42 patients with other urogenital diseases were analyzed using IMAC-Cu-3 ProteinChip. Proteomic spectra were generated by SELDI-TOF- MS. A preliminary "training" set of spectra derived from analysis of urine from 46 TCC patients, 32 patients with benign urogenital diseases (BUD), and 40 age-matched unaffected healthy men were used to train and develop a decision tree classification algorithm which identified a fine-protein mass pattern that discriminated cancers from non-cancers effectively. A blinded test set including 38 cases was used to determine the sensitivity and specificity of the classification system.

RESULTSThe algorithm identified a cluster pattern that, in the training set, segregated cancer from non-cancer with a sensitivity of 84.8% and specificity of 91.7%. The discriminatory pattern was correctly identified. A sensitivity of 93.3% and a specificity of 87% for the blinded test were obtained when compared the TCC versus non-cancers.

CONCLUSIONSELDI-TOF-MS technology is a rapid, convenient and high-throughput analyzing method. The urine tree analysis proteomic pattern as a screening tool is effective for differential diagnosis of bladder cancer. More detailed studies are needed to further evaluate the clinical value of this pattern.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Transitional Cell ; diagnosis ; urine ; Cystitis ; diagnosis ; urine ; Decision Trees ; Diagnosis, Differential ; Humans ; Male ; Middle Aged ; Prostatic Hyperplasia ; diagnosis ; urine ; Protein Array Analysis ; Proteomics ; methods ; Reproducibility of Results ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; Urinary Bladder Neoplasms ; diagnosis ; urine

OBJECTIVETo screen the gene expression profiles of IFN-alpha antiviral proteins based on a low-density cDNA Macroarray, and to explore the relationship between the expression of antiviral protein and the HBV replication.

METHODSThe HepG2 and HepG2.2.15 cells were treated with various concentrations of IFN-alpha (0 IU/ml, 100 IU/ml, 1000 IU/ml) of IFN-alpha for 6 h, and then the low-density cDNA Macroarray was used for analysing the expression profiles of antiviral genes and screening differential expressions of antiviral proteins. Meanwhile, the HepG2 cells were transiently transfected with HBV core protein-expressed plasmid pHBc-EGFP, and the expressions of antiviral proteins were analysed by RT-PCR assay. Moreover, the HepG2.2.15 cells were also transfected with the antiviral protein-expressed plasmid pcDNA3.1-Flag-MxA. ELISA was used for analysing the secreted HBV antigens, while dot blot and Southern blot were applied for analysing the extracellular HBV DNA and intracellular replicative intermediate HBV DNA in HepG2.2.15 cells. All data were presented as mean+/-SD and analyzed using the t-test and one-way analysis of variance (ANOVA) in the experiments.

RESULTSThe Macroarray results suggested that the expression of IFN-alpha antiviral genes like 6-16, IFITM1, IFITM2, IFITM3 and RING4 in HepG2.2.15 cells were partially inhibited. More importantly, it was found, in this research, the expression of antiviral protein MxA in HepG2.2.15 cells was completely suppressed. RT-PCR analysis indicated that the expression of MxA was also significantly decreased in HepG2 cells transfected with pHBc-EGFP plasmid. Although HepG2.2.15 cells transfected with pcDNA3.1-Flag-MxA plasmid could not inhibit extracellular HBV DNA and intracellular replicative intermediate HBV DNA, the MxA exerted some antiviral activities as it effectively suppressed the secretion of HBsAg and HBeAg in HepG2.2.15 cells.

CONCLUSIONSHBV and its antigen components probably influence the expression of antiviral proteins. IFN- resistance may be related to the down-regulation of antiviral proteins expression.

Antiviral Agents ; pharmacology ; Gene Expression Profiling ; Gene Expression Regulation, Viral ; Hep G2 Cells ; Hepatitis B virus ; drug effects ; physiology ; Humans ; Interferon-alpha ; pharmacology ; Plasmids

A comparison of recognition to migraine and treatment methods for it was made between traditional Chinese medicine and western medicine in this paper. Emphasis was made on standardized manipulation processes and techniques of sharp hooked needling for treating the disease, based on both the organic conception of the human body, the theory held by TCM, and on the principle of treatment according to pathogenesis obtained through differentiation of symptoms and signs. The necessary preparation before operations, methods for its application and announcements in the standardized manipulations of sharp hooked needling were all explained in detail. Based on its needling, pricking, dissecting and relaxing effects, the mechanisms of sharp hooked needling in treating the disease were investigated. It is concluded that sharp hooked needling is very effective in treating migraine.
Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; standards ; Humans ; Medicine, Chinese Traditional ; instrumentation ; methods ; standards ; Migraine Disorders ; therapy ; Needles ; Reference Standards