Comet Assay

UNLABELLEDOBJECTIVE To study the in vitro effect and mechanism of Ginkgo biloba Extract 50 (GBE50) for inhibiting beta-amyloid (Abeta)-induced oxidative stress in rats' hippocampal neurons.

METHODSThe primary hippocampal neurons were cultured in vitro and divided into 4 groups, i. e. the normal control group (Ctrl), the Abeta group, the propanediol control group (PDO), and the six GBE50 concentrations groups (5, 10, 25, 50, 100, and 200 microg/mL). Excepted the Ctrl group, neurons were induced to oxidative stress by 20 gmolLAbeta25-35. The MTT and fluorescent probes labeling were used to observe the effect of GBE50 with different concentrations on the cell viability and the generation of intracellular reactive oxygen species (ROS) in neurons. Furthermore, Western blot was used to detect the cytoplasmic/total cytochrome C (Cyto C) ratio and total intracytoplasmal Cyto C, and the effect of the expression of oxidative stress-related protein Cyto C and activated Caspase-3 in three GBE50 concentrations groups (25, 50, and 100 microg/mL).

RESULTSCompared with the Ctrl group, the cell vitality was obviously lowered and intracellular ROS generation significantly increased after induction of 20 micromol/L Abeta25-35 (both P < 0.05). Compared with the Abeta group, the cell vitality was evidently improved after treated with different GBE50 doses. Except for 10 microg/mL, the cell vitality could be obviously elevated along with increased drug concentrations (P < 0.05). Meanwhile, the intracellular ROS generation decreased significantly in each GBE50 dose groups (P < 0.05). Abeta could increase the cytoplasmic/total Cyto C ratio and enhance the activated Caspase-3 expression significantly (P < 0.05). Compared with the Abeta group, among the three concentrations of GBE50, the Cyto C ratio was obviously lowered in the 100 microg/mL GBE50 group (P < 0.05), and the expression of activated Caspase-3 significantly decreased in 50 microg/mL and 100 microg/mL GBE50 groups (P < 0.05).

CONCLUSIONS20 micromol/L Abeta25-35 could induce the generation of intracellular ROS in hippocampal neurons. GBE50 could inhibit Abeta induced intracellular oxidative stress of neurons through lowering the cytoplasmic/total Cyto C ratio and inhibiting the activation of apoptosis protein Caspase-3 expression.

Amyloid beta-Peptides ; toxicity ; Animals ; Cells, Cultured ; Cytochromes c ; metabolism ; Hippocampus ; metabolism ; Neurons ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Peptide Fragments ; toxicity ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley

To observe the reliability and short-term and mid-term efficacy of a new designed multi-functional lacrimal sac nasal drainage stent on the treatment of nasolacrimal duct obstruction after endonasal endoscopic dacryocystorhinostomy ( EES-DCR ) , and to evaluate its clinical application value. METHODS: A retrospective case series study was carried out. Seventy - six patients ( 79 eyes ) with nasolacrimal duct obstruction underwent EES-DCR and a new type of multi-functional lacrimal sac nasal drainage stent was implanted. The nasal lacrimal drainage stent was pulled out 2~3mo later after surgery and the patients received 3~6mo follow-up afterwards. RESULTS: ln 79 eyes, stents were successfully implanted in 77 eyes (97. 5%). All stents of 77 eyes could be pulled out successfully 2 ~ 3mo after the operation. Sixty eyes were cured with unobstructed lacrimal irrigation and symptom of epiphora disappeared;15 eyes were improved of epiphora and mild reflux during lacrimal irrigation were found, overall effective rate was 97. 4%, 2 eyes failed in stent implantation. ln 6mo follow-up, nocomplications such as stent displacement or slip was occurred, the epithelization of anastomotic stoma was good.CONCLUSlON: Surgical technique of using the multi-function nasolacrimal stent is simple and fast, and implantation of the new designed stents has a rapid recovery with fewer complications. The usage of the new stents not only improves the success rate but also broadens surgical indications of EES-DCR, it is a safe and effective method for the treatment of nasolacrimal duct obstruction.

Objective To assess the effectiveness of treatment of meniscal injuries of knee joints by arthroscopy.Methods 33 patients 35 joints were followed up and the parts,types and treatment under arthroscopy were analysed.Results 33 patients were followed up from six months to six years,the mean preoperative Lysholm score was 60.5 points,and the mean postoperative one was 86.7 points.Conclusion The advantage of treating meniscal injuries by arthroscopy was the result of correct examination and little wound of arthroscopy operation,and arthroscopic repair or partial menisectomy could effectively restore the function of the injured knee.

Objective To study the mRNA expression of HS1-associated protein X-1(Hax-1),an an- ti-apoptosis genc,in the peripheral blood mononuclear cells(PBMC)of patients with systemic lupus erythe- matosus(SLE),and further investigate the roles and significance of Hax-1 in the pathogenesis of SLE.Meth- ods Generation of longer cDNA fragments from serial analysis of gene expression(SAGE)tags for gene identi- fication(GLGI)was applied to identify the gene Hax-1 according to the Long SAGE tag.Then reverse tran- scription-polymerase chain reaction(RT-PCR)technique was used to semiquantitatively analyze mRNA ex- pressions of Hax-1 in PBMC from 34 active SLE patients and 25 healthy subjects.Results Compared with healthy controls,there was significant difference between SLE patients in the active stage and the normal controls(Z=-4.556,P＜0.01).The average level of mRNA expression in active SLE group was higher than that in healthy controls.Significant difference was found between the group with mild SLE and either the moderate or the severe one(P＜0.01).Conclusion The mRNA expression level of Hax-1 in active SLE group increase markedly,and to some extent,it is related to the activity of SLE.This provides a valuable basis for the further study on the role of apoptosis in SLE.

Background: Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are X?linked recessive neuromuscular diseases resulting from dystrophin (DMD) gene mutations. It has been known that the carrier of DMD mutations may also have symptoms of the disease. While de novo mutation is quite common in BMD/DMD patients, it is rarely reported in the female carriers. Methods: Two sporadic Chinese patients with progressive muscular dystrophy and their familial members were recruited. The targeted next?generation sequencing (NGS) and the multiplex ligation?dependent probe analysis (MLPA) were performed in the proband. Blood tests, electrocardiography, echocardiography, and electromyography were also evaluated. Results: Two novel mutations of DMD gene were identified, c.7318C>T(p.Q2440*) in the male proband and c.4983dupA(p.A1662Sfs*24) in the female carrier. The MLPA analysis did not detect any large rearrangements. The haplotype analysis indicated that the two mutations were derived from de novo mutagenesis. Conclusions: We identified two novel de novo mutations of DMD gene in two Chinese pedigrees, one of which caused a female patient with muscular dystrophy. The mutational analysis is important for DMD patients and carriers in the absence of a family history. The NGS can help detect the mutations in MLPA?negative patients.

In order to study the role of PML-RARalpha fusion gene and its expression product during apoptotic process in NB4 cells induced by arsenic trisulfide (As(2)S(3)), the apoptotic effects of NB4 cells were observed by cell morphology, flow cytometry and DNA electrophoresis. The change of PML-RARalpha fusion gene and its expression product were also assayed by chromosomal G banding, RT-PCR and Western blot. The results showed that arsenic trisulfide induced apoptosis of NB4 cells, during this process, PML-RARalpha fusion gene had no significant changes, but the expression of PML-RARalpha fusion protein and wild-type RARalpha were all reduced. It is concluded that arsenic trisulfide can induce apoptosis of NB4 cells, the degradation of PML-RARalpha fusion protein and wild-type RARalpha may play an important role during apoptotic process.
Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Blotting, Western ; Cell Division ; drug effects ; Dose-Response Relationship, Drug ; Flow Cytometry ; Humans ; Neoplasm Proteins ; genetics ; physiology ; Oncogene Proteins, Fusion ; genetics ; physiology ; RNA, Neoplasm ; drug effects ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sulfides ; pharmacology ; Tumor Cells, Cultured

The vitamin K epoxide reductase complex subunit 1 (VKORC1), the rate-limiting enzyme for vitamin K recycling, is significantly down-regulated in the kidneys of urolithiasis patients. This study searched for direct evidence to define the inhibitory activity of VKORC1 against calcium oxalate (CaOx) crystal formation. In the experiment of VKORC1 overexpression, HK-2 cells were transfected with the pFLAG-CMV-7.1-VKORC1 plasmid as a pFLAG-CMV-7.1-VKORC1 transfection group or the pFLAG-CMV-7.1 plasmid as a pFLAG-CMV-7.1 control group. In the experiment of VKORC1 knockdown, HK-2 cells were transfected with the PGPU6/GFP/Neo-VKORC1shRNA-2 as a PGPU6/GFP/Neo-VKORC1shRNA-2 transfection group or the PGPU6/GFP/Neo-shRNA-NC plasmid as a PGPU6/GFP/Neo-shRNA-NC control group. The expression of VKORC1 in HK-2 cells was detected by real-time quantitative PCR and Western blotting. The CaOx crystal formation was observed under the laser-scanning confocal microscope. It was found that the expression levels of VKORC1 mRNA and protein were significantly higher in the pFLAG-CMV-7.1-VKORC1 transfection group than in the pFLAG-CMV-7.1 control group (P<0.01). The number of CaOx crystals in HK-2 cells incubated in fluorescently labeled CaOx monohydrate (COM) crystal medium for 48 h was 14±4 per field (100×) in the pFLAG-CMV-7.1-VKORC1 transfection group and 26±5 per field (100×) in the pFLAG-CMV-7.1 control group respectively under the laser-scanning confocal microscope. The amount of CaOx crystal aggregation and formation in the pFLAG-CMV-7.1-VKORC1 transfection group was significantly reduced as compared with the pFLAG-CMV-7.1 control group (P<0.05). The expression levels of VKORC1 mRNA and protein were significantly lower in the PGPU6/GFP/Neo-VKORC1shRNA-2 transfection group than in the PGPU6/GFP/Neo-shRNA-NC control group (P<0.05). The number of CaOx crystals in HK-2 cells incubated in fluorescently labeled COM crystal medium was 65±11 per field (100×) in the PGPU6/GFP/Neo-VKORC1shRNA-2 transfection group and 24±6 per field (100×) in the PGPU6/GFP/Neo-shRNA-NC control group respectively under the laser-scanning confocal microscope. The amount of CaOx crystal aggregation and formation in the PGPU6/GFP/Neo-VKORC1shRNA-2 transfection group was significantly increased as compared with the PGPU6/GFP/Neo-shRNA-NC control group (P<0.05). These findings suggested that the VKORC1 protein could inhibit CaOx salt crystallization, adhesion and aggregation. This research would help us to understand the mechanisms involving the interaction between crystallization and epithelial cells and the formation of CaOx.

Tadalafil is an oral phosphodiesterase type-5 inhibitor for male erectile dysfunction (ED). It has been proved to improve erectile function, with safety and effectiveness lasting up to 36 h, allowing patients to choose when to have sexual activities. Tadalafil is also efficacious in the treatment of ED associated with such diseases as hypertension, diabetes mellitus, etc. This article reviews the available evidence for the efficacy of tadalafil on ED and related diseases.
Carbolines ; therapeutic use ; Diabetes Mellitus ; drug therapy ; physiopathology ; Erectile Dysfunction ; drug therapy ; physiopathology ; Humans ; Hypertension ; drug therapy ; physiopathology ; Male ; Phosphodiesterase Inhibitors ; therapeutic use ; Tadalafil ; Treatment Outcome

Adult ; Biopsy ; DNA ; genetics ; Female ; Gene Expression Profiling ; Glomerulonephritis, IGA ; genetics ; pathology ; Humans ; Kidney ; pathology ; Male ; Middle Aged ; Young Adult