AIM:To investigate the effects of ginsenoside Rg1 on the behavioral changes and the autophagy of hippocampal neurons of the rats with post-traumatic stress disorder (PTSD).METHODS:The Sprague-Dawley rats were randomly divided into 5 groups:control group, model group, fluoxetine group, low-dose ginsenoside Rg1 group and high-dose of ginsenoside Rg1 group.The combination of single prolonged stress and foot stock was performed to induce PTSD-like animal model.The rats in fluoxetine group was administered with fluoxetine by gavage at dose of 10 mg/kg for 21 d, while the rats in low and high doses of ginsenoside Rg1 groups were administered with ginsenoside Rg1 by gavage at doses of 20 mg/kg and 40 mg/kg for 21 d, respectively.The rats in control group and model group were both given saline by gavage for 21 d.The open-field test and stiff behavior test were used to examine the behavioral changes of the rats.The morphological structure and numerical changes of the hippocampal neurons were observed by Nissl-staining method.We adopted immunofluorescence labeling to observe the beclin 1 and LC3 positive hippocampal neurons and the levels of beclin 1 and LC3-Ⅱ/LC3-Ⅰratio in rat hippocampus.RESULTS:Compared with control group, decreased vertical movement time and horizontal movement time in open-field test and increased rate of stiff behavior in the stiff behavior test were observed in model group.Hippocampal neurons in model group were loosely arranged with vacuole-like structures and different degrees of cell shrinkage in contrast with control group.More beclin 1 and LC3 positive cells were identified, and higher protein levels of beclin 1 and ratio of LC3-Ⅱ/LC3-Ⅰ in model group were found as compared with control group.However, increase in movement in open-field test and decrease in stiff behavior were detected in the rats treated with low-and high-dose ginsenoside Rg1 as compared with the model rats.Meanwhile, vacuole structures, the numbers of beclin 1 and LC3 positive neurons, the protein expression of beclin 1 and LC3, and the total cell numbers were increased.Higher dose of ginsenoside Rg1 had more profound effects on these observed results.CONCLUSION:Ginsenoside Rg1 alleviates the abnormal behaviors in the PTSD rats, which might be related to the inhibition of abnormal autophagy of hippocampal neurons.

Objective To investigate the role of acid and bile reflux in children with gastroesophageal reflux disease (GERD) and to evaluate the significance of detecting acid and bile reflux in diagnosing GERD in children.Methods Using ambulatory 24 h pH mo-(nitoring) and bilirubin monitoring technique, we simultaneously assessed the changes of intraesophageal pH and bile reflux in 23 subjects (including 11 healthy controls and 12 patients with GERD).Results The time of esophageal acid exposure (pH

Human papilloma virus (HPV) infection is closely correlated with the occurrence of cervical cancer and precancerous lesion, high risk HPV can induce cervical cancer by the corresponding protein of E6 and E7 gene expression. Earlier diagnosis and earlier prevention of HPV infection are the key points in blocking cervical cancer, and combined use of Chinese herbal preparations with Western medicine is the important way for preventing HPV infection and prohibiting cervical canceration. So, to develop anti-viral chemical compound from natural drugs has become the hotspot of research today.
Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interferons ; therapeutic use ; Oncogene Proteins, Viral ; analysis ; Papillomavirus E7 Proteins ; analysis ; Papillomavirus Infections ; drug therapy ; Phytotherapy ; Repressor Proteins ; analysis ; Uterine Cervical Neoplasms ; drug therapy ; virology

For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; Hepatic Stellate Cells ; High-Throughput Screening Assays ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Plasmids ; Promoter Regions, Genetic ; RNA, Messenger ; Transfection ; Transforming Growth Factor beta1 ; pharmacology

Objective To investigate the effect of solute cartier organic anion transporter family, member 1B1 (SLCOIBI) gene variants on the response to therapy with repaglinide in type 2 diabetes. Methods 100 newly-diagnosed type 2 diabetic patients were treated with repaglinide during a course of 48 weeks. Anthropometrie parameters and indices related to glucose metabolism were measured periodically. Genotypes of SLCO1B1 D130N and V174A were detected by PCR-restriction fragment length polymorphism (RFLP) and sequencing respectively. Results Eighty-nine patients accomplished the 48-week follow-up visits. D130N variant in SLCO1B1 gene was associated with repaglinide treatment, DD genotype had better HbA1C lowering effect than N allele carrier [△HbA1C: (-2.29±0.23) % vs (-1.49±0.21)%, P<0.05]. No association was detected between D130N and the other effects of repaglinide on glucose metabolism related phenotypes. Conclusion D130N variant in SLCO1B1 gene is associated with the response to repaglinide treatment in patients with type 2 diabetes. DD homozygotes had a better effect than N allele carriers.

Objective To establish real-time PCR with SYBR Green Ⅰ for quantification the gene of survivin.Methods The components and conditions of PCR system were optimally determined by fluorescence intensity,cycle threshold(Ct),melting curve,coefficiency and slope of the standard curve.The means to eliminate the contaminating fluorescence of primer-dimers and the mode for Ct value determination were also optimized.Using the developed PCR system,we quantificated the survivin gene in 43 patients with gastric carcinoma.Results The optimized condition for PCR amplification of survivin were 2 mmol/L of MgCl_2,2.5 U/100 ?l of Taq DNA polymerase,0.2 ?mol/L of primers,and the optimized annealing temperatures for PCR were 58℃.The influence of primer dimmer can be eliminated by setting the fluorescence collecting temperature below the Tm of the specific amplicon by 2℃.The second derivative maximum mode,instead of fit point mode,was a feasible method to determine the Ct value for quantification.The sensitivity of this method was 10 copies/?l,and a good linearity was found from 10~1 to 10~4 copies/?l(r = 0.999 7).The inter-experimental coefficient of variation was 1.13%-1.91%,whereas the coefficient of variation between runs was 3.31%-4.50%.Using the optimized PCR system,we quantificated the gene of survivin,the result indicated that survivin gene was amplified in 13.9% of gastric carcinomas.Conclusions The optimal real-time PCR with SYBR Green Ⅰ,as a cost-effective and feasible DNA quantitative method,is fit for quantification of the survivin with satisfactory repeatability and high sensitivity.

Objective To investigate the nerve recanalization and the motor function of hind legs after transplantation of peripheral nerve grafts treated with microsurgical technique at chronic spinal cord injury (SCI) in rats.Methods The SD rats were established into SCI model with improved Allen method.The rats were divided into two parts 12 weeks after the injury.In experimental group:by microsurgieal technique. the sural nerves were removed epineufium and transplanted into SCI lesion,control group rats were treated without any operation.Retrograde HRP tracing through sciatic nerve were practiced at 1 month,2 month,3 month after transplantation of peripheral nerve grafts.The morphological changes were observed at section of spinal cord and the motor functions of both hind legs of rat were detected.Results The morphology of the injured spinal cord sections turned better.Retrograde HRP tracing through sciatic nerve showed some HRP positive markers at the site of near rostral end of the nearly injured part at one month after transplantation and increased with the time going by.Motor function of hind legs of rats recovered significantly in transplantation groups.Conclusion Peripheral nerve grafts treated with mierosurgical technique have repairing effect on chronic spinal cord injury in rats.

BACKGROUND:Unlike linear RNAs,circular RNAs (circRNAs) are a novel type of RNA which can form covalently closed circles and are highly expressed in eukaryotic transcriptomes.In the plenitude of naturally occurring RNAs,circRNAs and their biological role are underestimated for years.Recent studies have discovered thousands of endogenous circRNAs in mammalian cells.OBJECTIVE:To review the formation,properties,and functions of circRNAs,and their potential significance in diseases.METHODS:A computer-based search for literature in CNKI and PubMed databases published from January 2000 to December 2016 was performed using the keywords of "circRNA,miRNA,function,mechanism" in English and Chinese,respectively.Finally,62 eligible articles were included for analysis.RESULTS AND CONCLUSION:CircRNAs are largely generated from exonic or intronic sequences,and reverse complementary sequences or RNA-binding proteins are necessary for circRNA biogenesis.A majority of circRNAs that are conserved across specie are stable and resistant to RNaseR,and often exhibit tissue/developmental-stage-specific expression.Recent research has revealed that circRNAs can function as microRNA sponges,regulators of splicing and transcription,and modifiers of parental gene expression.Emerging evidence indicates that circRNAs might play important roles in atherosclerotic vascular diseases,neurological disorders,prion diseases and cancer;exhibit aberrant expression in colorectal cancer;and serve as diagnostic or predictive biomarkers of some diseases.Similar to microRNAs and long noncoding RNAs,circRNAs are arousing general interest in the field of RNA and widely participate in the life process.

OBJECTIVETo assess the ability of tetrandrine (Tet) to enhance the sensitivity to irradiation and its mechanism in cell lines of human breast cancer p53-mutant MCF-7/ADR, p53-wild-type MCF-7 and human colon carcinoma p53-mutant HT-29 as well as in C26 colorectal carcinoma-bearing BALB/c mice.

METHODSMCF-7/ADR, HT-29 and MCF-7 cells were exposed to irradiation in the absence or presence of tetrandrine. The effect of Tet on the cytotoxicity of X-irradiation in these three cells was determined and the effect of tetrandrine on cell cycle arrest induced by irradiation in its absence or presence was studied by flow cytometry. Moreover, mitotic index measurement determined mitosis of cells to enter mitosis. Western blotting was employed to detect cyclin B 1 and Cdc2 proteins in extracts from irradiated or non-irradiated cells of MCF-7/ADR, HT-29 and MCF-7 treated with tetrandrine at various concentrations. Tumor growth delay assay was conducted to determine the radio-sensitization of tetrandrine in vivo.

RESULTSClonogenic assay showed that tetrandrine markedly enhanced the lethal effect of X-rays on p53-mutant MCF-7/ADR and HT-29 cells and the sensitization enhancement ratio (SER) of tetrandrine was 1.51 and 1.63, but its SER was only 1.1 in p53-wt MCF-7 cells. Irradiated p53-mutant MCF-7/ADR and HT-29 cells were only arrested in G2/M phase while MCF-7 cells were arrested in G1 and G2/M phases. Radiation-induced G2 phase arrests were abrogated by tetrandrine in a concentration-dependent manner in MCF-7/ADR and HT-29 cells, whereas redistribution within MCF-7 cell cycle changed slightly. The proportion of cells in M phase increased from 1.3% to 14.7% in MCF-7/ADR cells, and from 1.5% to 13.2% in HT-29 cells, but 2.4% to 7.1% in MCF-7 cells. Furthermore, the levels of cyclin B 1 and Cdc2 expression decreased after X-irradiation in MCF-7/ADR and HT-29 cells, and the mitotic index was also lower. Tet could reverse the decrease and induce the irradiated cells to enter mitosis (M phase). Endosomatic experiment showed that tetrandrine caused tumor growth delay in irradiated mice.

CONCLUSIONTetrandrine boosts the cell killing activity of irradiation both in vitro and in vivo. Tetrandrine is a potent abrogator for G2 checkpoint control and can sensitize the cells to radiation.

Animals ; Benzylisoquinolines ; pharmacology ; CDC2-CDC28 Kinases ; metabolism ; Cell Line, Tumor ; Cyclin B ; metabolism ; Cyclin B1 ; Drug Screening Assays, Antitumor ; G2 Phase ; drug effects ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Radiation Tolerance

OBJECTIVES: We evaluate the long-term surgical and hearing results using a canal wall down mastoidectomy technique followed by cavities obliteration, canal wall reconstruction and ossiculoplasty without staging. METHODS: A total of 44 patients between January 2002 and October 2009 were selected and 27 of them were revision cases. Preoperative and postoperative pure tone average (PTA) and air-bone gap (ABG) were assessed and compared 1 and 4 years after surgery. RESULTS: The middle ear was well healed and aerated in 40 patients (90.9%) and the tympanic membrane was intact in 42 patients (95.5%). Recurrent cholesteatoma was found on postoperative follow-up in two of the revision patients (7.4%) but none in the primary patients. Seven patients were found to have partial canal bone absorption, but revision surgery was not required. Over 86.4% of all cases were water resistant. Long-lasting improvement and/or preservation of hearing, with maintenance of PTA-ABG closure in 63.7% of all cases within 20 dB, were obtained. CONCLUSION: The efficacy of our technique after a canal wall down mastoidectomy is satisfactory, and the rate of complication is acceptably low. We believe that our technique could be a convenient method in disease control and providing an excellent basis for hearing restoration simultaneously.
Absorption ; Cholesteatoma ; Ear, Middle ; Follow-Up Studies ; Hearing ; Humans ; Mastoid ; Reconstructive Surgical Procedures ; Surgical Procedures, Operative ; Tympanic Membrane ; Water