Objective:To study the effect of metformin on proliferation,cell cycle and apoptosis of U937 cells.Methods: U937 cells were treated with different concentrations of metformin,collected cells in 24,48 and 72 hours.Subsequently,cell proliferation was assessed by CCK-8 assay,and the cell cycle and apoptosis were analyzed by flow cytometry (FCM).The expression of Bcl-2,Bax,p-AMPK,p53 were determined by Western blot.Results: The proliferation of U937 cells was inhibited by metformin in a time-and dose-dependent manner.Metformin-treated cells were arrested at G0/G1 phase,the cell frequency at G0/G1 phase was increased in a time-and dose-dependent manner.Metformin also induced cell apoptosis in a dose-dependent manner.It showed that 20 mmol/L metformin induced cell apoptosis in a time-dependent manner.The expression of p-AMPK,p53,Bax was up-regulated while Bcl-2 expression was down-regulated after metformin treatment.Conclusion: Metformin could inhibit the U937 cell proliferation,block the cell cycle at G0/G1 phase,and induce cell apoptosis,which may partially be attribute to the up-regulation of Bax,down-regulation of Bcl-2,activation of AMPK/p53 signaling.

The purpose of the present study is to observe the effect of electro-acupuncture (EA) stimulation on intracerebral neurotransmitters in a rat model of Parkinson's disease (PD), and explore the possible mechanism. We used 6-hydroxydopamine (6-OHDA) injection in medial forebrain bundle (MFB) in the right brain of Sprague Dawley (SD) rat to establish the parkinsonian rat model, and randomly divided the PD rats into model and 100 Hz EA stimulation groups (n =10 in each group). EA stimulation group received 4 courses of EA stimulation on Baihui (GV-20) and Dazhui (GV-14) acupuncture points. Moreover, ten rats were randomly selected as sham operation group, only receiving normal saline (NS) injection in MFB. Then apomorphine (APO)-induced rotational behavior in different groups was recorded, and the contents of γ-aminobutyric acid (GABA) in the brain were analyzed with high pressure/performance liquid chromatography-electrochemical detection (HPLC-ECD). The results showed that model group exhibited abnormal rotational behavior with APO treatment, suggesting the successful establishment of PD model. Compared with sham operation group, model group showed increased GABA contents in cortex and striatum, as well as decreased GABA content in ventral midbrain, on the lesioned side. EA stimulation could effectively ameliorate the abnormal rotational behavior of PD rat. Compared with the model group, EA stimulation decreased the ratio of GABA content on the lesioned side to that on unlesioned side in the cortex, while increased the ratios in the striatum and cerebellum. However, there was no difference of the ratio in the ventral midbrain among three groups. These results suggest high-frequency EA stimulation significantly improves the abnormal behavior of PD rats, which may exert through enhancing the inhibitory effect of cerebellum-basal ganglia-cortical loop on motor center.
Acupuncture Therapy ; Animals ; Brain ; metabolism ; Electroacupuncture ; Male ; Motor Cortex ; physiology ; Oxidopamine ; Parkinson Disease, Secondary ; chemically induced ; metabolism ; therapy ; Rats ; Rats, Sprague-Dawley ; gamma-Aminobutyric Acid ; metabolism

Cholelithiasis is one of the clinically common and frequently encountered diseases. In this paper, the Chinese Meteria Medica and prescriptions utilized to treat cholelithasis were discussed in four aspects. In addition, we discussed the clinical effect and mechanism of actions of these drugs in order to provide some reference for future drug development in this area.
Animals ; Cholelithiasis ; drug therapy ; prevention & control ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Humans ; Phytotherapy ; Plants, Medicinal ; chemistry

Objective To investigate the value of 18F-FDG/99Tcm-MIBI SPECT myocardial imaging for the detection of myocardial viability and prognosis in patients with AMI. Methods 18F-FDG/99Tcm-MIBI SPECT myocardial imaging was performed in 98 consecutive patients [man 87, women 11; average age (58 ±11)y] with AMI. The myocardium was scored individually for nine segments: mildly decreased uptake = 1,significantly decreased uptake = 2, and no uptake = 3. Perfusion defect but preserved 18 F-FDG uptake was defined as perfusion-metabolism mismatch, indicating jeopardized but viable myocardium. Perfusion defect and decreased 18 F-FDG uptake were defined as match, indicating myocardial necrosis. Echocardiogram was performed before and after treatment for evaluating the LVEF. All patients were followed after treatment.The rate of cardiac events was calculated and compared between patients with medication and revascularization. Paired t test, Chi-square test and log-rank test were used for statistical analysis. Results In the group with viable myocardium, 27 patients received revascularization and 10 received medication. In the group with infarcted myocardium, 26 patients received medication and 35 received revascularization. Patients underwent revascularization and with medication had no significant difference in improvement of LVEF between both groups (viable myocardium group: χ2 = 0.509, P > 0. 05; infarcted myocardium group: χ2 =0.035, P > 0.05). In viable myocardium group, cardiac event rate was significantly higher in patients with medication than in those who had undergone revascularization (50.0% vs 14.8%, χ2 =4.91, P<0.05).In the infarcted myocardium group, cardiac event rate was also significantly higher in patients with medication (30.7% vs5.7% ,χ2 =6.83, P<0.05). Conclusions 18F-FDG/ -MIBI SPECT myocardial imaging may well be of value but limited for the detection of myocardial viability and prediction of improvement in cardiac function as well as prognosis. However, more prospective data are needed for final evaluation.

OBJECTIVETo investigate the association of male infertility with single nucleotide polymorphisms of the autosomal DAZL gene.

METHODSSemen samples were collected from infertile men (infertility group, n=144) and healthy fathers (fertility group, n=53) in the northeast of China, and the former was further divided into 5 subgroups according to the WHO diagnostic criteria for oligoasthenoteratozoospermia (1999) i.e., oligoasthenoteratozoospermia, oligoasthenozoospermia, asthenoteratozoospermia, asthenozoospermia and non-oligoasthenoteratozoospermia. PCR-RFLP analysis was used to detect the polymorphism of SNP260. The SNP386 polymorphism of the DAZL gene was analysed with a PCR-SSCP based method. Sequencing and statistical analyses were performed.

RESULTSThe SNP260 polymorphism of the DAZL gene was found in both the infertility and the control groups, with no significant difference in between (P > 0.05). There was a larger percentage of SNP260AG in the subgroup of oligoasthenoteratozoospermia than in others, and the SNP386 polymorphism of the DAZL gene was not found in any of the subgroups.

CONCLUSIONThe SNP260 and SNP386 polymorphisms of the DAZL gene are not associated with male infertility in the northeast of China. Further research needs to be done to clarify the association between SNP260AG and oligoasthenoteratozoospermia. SNP386 polymorphism may be restricted in some small area in Taiwan only. Therefore neither of them can be used as a molecular marker for the genetic diagnosis of male infertility in the northeast of China.

Adult ; Base Sequence ; China ; DNA Mutational Analysis ; Humans ; Infertility, Male ; genetics ; Male ; Oligospermia ; genetics ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Polymorphism, Single-Stranded Conformational ; RNA-Binding Proteins ; genetics ; Taiwan

OBJECTIVETo compare the effectiveness of immunofluorescence and polymerase chain reaction (PCR) in detecting human papilloma virus (HPV) in condyloma acuminata (CA).

METHODSHPVs in CA tissues from 60 patients were detected by immunofluorescence and PCR, respectively. Different subtypes of HPVs were also identified with restriction fragment length polymorphism (RFLP).

RESULTSThe positive detective rates of immunofluorescence and PCR were 56.67% (34/60) and 96.67% (58/ 60), respectively (P < 0.01). RFLP results showed HPV6 and HPV11 were the main subtypes in the detected virus, which accounted for 98.28%.

CONCLUSIONThe sensibility of PCR is superior to that of immunofluorescence.

Condylomata Acuminata ; virology ; Female ; Human papillomavirus 11 ; genetics ; isolation & purification ; Human papillomavirus 6 ; genetics ; isolation & purification ; Humans ; Male ; Microscopy, Fluorescence ; Polymerase Chain Reaction

Biomacromolecules play an important role in the treatment of many diseases, but as a result of cell membrane serving as the natural barriers, only the small molecular compounds whose molecular weights are smaller than 600 Da can get through cell membrane and enter the cell. In recent years, some short peptides (the length less than 30 amino acids) are found to have the cell-penetrating function, called cell-penetrating peptides (CPPs). They are able to effectively translocate segments of protein, polypeptides, nucleic acid into the cells of many mammal animals with many methods. They have high transduction efficiency and will not lead to cell damage. So, the discovery of CPPs has a very good applicable prospect in such research fields as cell-biology, gene-therapy, drug transduction in vivo, evaluation of clinical medicine and medical immunology. This paper reviews the types and characteristics of CPPs, internalization mechanisms, applications, and their existing problems.
Absorption ; drug effects ; Amino Acid Sequence ; Animals ; Cell Membrane Permeability ; Cell-Penetrating Peptides ; classification ; pharmacology ; physiology ; Drug Carriers ; Endocytosis ; physiology ; Humans ; Protein Transport

This study was to investigate the changes of autonomic nerve function and hemodynamics in patients with vasovagal syncope (VVS) during head-up tilt-table testing (HUT).HUT was performed in 68 patients with unexplained syncope and 18 healthy subjects served as control group.According to whether bradycardia,hypotension or both took place during the onset of syncope,the patients were divided during the test into three subgroups:vasodepressor syncope (VD),cardioinhibitory syncope (CI) and mixed syncope (MX) subgroups.Heart rate,blood pressure,heart rate variability (HRV),and deceleration capacity (DC) were continuously analyzed during HUT.For all the subjects with positive responses,the normalized low frequency (LFn) and the LF/HF ratio markedly decreased whereas normalized high frequency (HFn) increased when syncope occurred.Syncopal period also caused more significant increase in the power of the DC in positive groups.These changes were more exaggerated compared to controls.All the patients were indicative of a sympathetic surge in the presence of withdrawal vagal activity before syncope and a sympathetic inhibition with a vagal predominance at the syncopal stage by the frequency-domain analysis of HRV.With the measurements ofDC,a decreased vagal tone before syncope stage and a vagal activation at the syncopal stage were observed.The vagal tone was higher in subjects showing cardioinhibitory responses at the syncopal stage.DC may provide an alternative method to understand the autonomic profile of VVS patients.

AIMTo identify a novel alternative transcript of the novel retinal pigment epithelial cell gene (NORPEG) expressed in the human testis.

METHODSA human testis cDNA microarray was established and hybridized with cDNA probes from human fetal testes, adult testes and human spermatozoa. Differentially expressed clones were sequenced and analyzed. One of these clones was a short transcript of NORPEG which we proceeded to analyze by RT-PCR.

RESULTSThe novel short alternative transcript of NORPEG was isolated and named sNORPEG. It was 3486 bp in length and contained a 2952-bp open reading frame, encoding a 110.4-kDa protein of 983 amino acids. Amino acid sequence analysis showed that the sNORPEG protein contains six ankyrin repeats and two coiled-coil domains. It shares a high homology with the NORPEG and ankycorbin proteins in both its sequence and motifs. Blasting the human genome database localized sNORPEG to human chromosome 5p13.2-13.3. Expression profiles showed that sNORPEG was expressed in human fetal testes, adult testes and spermatozoa. Moreover, sNORPEG was found to be ubiquitously expressed in human tissues.

CONCLUSIONsNORPEG is expressed in different developmental stages of the testis and encodes a protein that may have roles in human testis development and spermatogenesis.

Alternative Splicing ; Amino Acid Sequence ; Base Sequence ; Cytoskeletal Proteins ; genetics ; DNA, Complementary ; Gene Expression Profiling ; Humans ; Male ; Molecular Sequence Data ; Open Reading Frames ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology, Amino Acid ; Testis ; metabolism ; Transcription Factors ; genetics

AIMTo identify genes related to the human testis development by substrate hybridization technique.

METHODSA human testis cDNA microarray was constructed and hybridized with probes prepared from human adult and fetal testes and spermatozoa mRNAs by reverse transcription reactions. The differentially expressed genes were sequenced. And a newly identified cullin-3 (CUL-3) transcript variant (designated cul-3b) was bio-informatically analyzed with an online GenBank database. Multi-tissue reverse transcription polymerase chain reaction (RT-PCR) was used to determine the tissue expression profile of cul-3b.

RESULTSCul-3b, a novel CUL-3 transcript variant, was identified. The expression level of cul-3b in adult testes was 3.79-fold higher than that in fetal ones. Cul-3b differed from cul-3 (including NM_003590 and AY337761) in the opening reading frame and had three internal ribosomal entry sites IRESes in the 5'-UTR. These led to a 24 amino acid (aa) truncation at N-terminus of CUL-3b as compared with CUL-3 and a more motivated expression pattern of cul-3b under some strict circumstances. Additionally, cul-3b expressed ubiquitously in human tissues according to multi-tissue RT-PCR.

CONCLUSIONCul-3b is a novel transcript variant of CUL-3, which may be important not only for the development of human testis but also for that of other organs.

Base Sequence ; Cell Cycle Proteins ; genetics ; Cullin Proteins ; genetics ; Humans ; Molecular Sequence Data ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction