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

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.

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.

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 investigate the effect of rabbit bone marrow mesenchymal stem cells (MSCs) as seed cells for the repair of tendon defect.

METHODSThe MSCs were isolated, amplified and identified by detection of surface protein CD44 mRNA. A 3 cm long defect was made in the Achilles tendon of the rabbit. The rabbits were divided into experimental (E) and control (C) groups. The autologous MSCs were implanted into a collagen-polyglycolic acid (PGA) scaffold to form a tissue-engineered tendon, which was then transplanted to bridge the defect in the E group, while only collagen-PGA was transplanted to bridge the defect in the C group. The transplanted tendon was observed grossly and microscopically at 4, 8, 12 weeks after operation.

RESULTSThe cultured MSCs exhibited positive staining of CD44 on 11 days after in vitro culture. A tendon-like tissue could be discerned at the operation site in the E group 4 weeks after operation. Tendon-like cells similar to normal tendon tissue, being axially arranged in collagen matching the mechanical direction, with uniform morphology could be seen in E group 12 weeks after operation. The newly regenerated tissue in C group adhered to the adjacent tissue and was smaller than that in E group. The collagen fibers in the regenerated tissue were loose with reticular and filiform structure, and the cells were arranged disorderly 12 weeks after the transplantation.

CONCLUSIONIt is feasible to repair the tendon defect with autologous MSCs as seed cells.

Achilles Tendon ; injuries ; Animals ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Mesenchymal Stem Cell Transplantation ; Rabbits ; Tendon Injuries ; surgery ; Tissue Engineering ; methods

OBJECTIVETo compare the clinical effects and radiographic outcomes of mini-open trans-spatium intermuscular and percutaneous short-segment pedicle fixation in treating thoracolumbar mono-segmental vertebral fractures without neurological deficits.

METHODSFrom August 2009 and August 2012, 95 patients with thoracolumbar mono-segmental vertebral fractures without neurological deficits were treated with short-segment pedicle fixation through mini-open trans-spatium intermuscular or percutaneous approach. There were 65 males and 30 females, aged from 16 to 60 years old with an average of 42 years. The mini-open trans-spatium intermuscular approach was used in 58 cases (group A) and the percutaneous approach was used in 37 cases (group B). Total incision length, operative time, intraoperative bleeding, fluoroscopy, hospitalization cost were compared between two groups. Visual analog scale (VAS) and radiographic outcomes were compared between two groups.

RESULTSAll patients were followed up from 12 to 36 months with an average of 19.6 months. No complications such as incision infection, internal fixation loosening and breakage were found. In group A, fluoroscopy time was short and hospitalization cost was lower than that of group B (P<0.05). But the total incision length in group B was smaller than that of group A (P<0.05). There was no significant differences in operative time, intraoperative bleeding, postoperative VAS and radiographic outcomes between two groups (P>0.05). Postoperative VAS and radiographic outcomes were improved than that of preoperative (P<0.05).

CONCLUSIONThe mini-open trans-spatium intermuscular and percutaneous short-segment pedicle fixation have similar clinical effects and radiographic outcomes in treating thoracolumbar mono-segmental vertebral fractures without neurological deficits. However, in this study, the mini-open trans-spatium intermuscular approach has a short learning curve and more advantages in hospitalization cost and intraoperative radiation exposure times, and is recommendable.

Adolescent ; Adult ; Female ; Fluoroscopy ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries ; surgery ; Visual Analog Scale