Objective Spermatogonial stem cells(SSCs)dissociated from 2～5 days postpartum mice were cultured on Sertoli cells feeders,to study the effect of Sertoli cells feeders on culture of mouse spermatogonial stem cells.Methods Specific markers CD9 of mouse SSCs cultured serum-free StemPro-34 SFM culture medium were identified by immunohistochemical assay.Result During the first week of culture,on Sertoli cells feeders or STO feeders,the biologica1 behaviors of spermatogonial stem cells showed no obvious difference.After a week of culture,compared with control,there were more number of spermatogonial stem cells remained when they were cultured for 60 days.These cells were expressed CD9 positive.In conclusion Sertoli cells can be used as feeders not only to promote survival but also renewal of spermatogonial stem cells.

Objective To explore the effect of long-term chronic mild stress on depression-like behaviors and the expression of heat shock protein 70(HSP70) in rats.Methods The standard SD rats were divided into two groups as following:one group (n =6) was treated with chronic mild stress for 3 weeks,and another group (n =5) was treated with chronic mild stress for 6 weeks.Depression-like behaviors of rats was observed by sucrose consumption test and open field test before and after chronic mild stress.And Western Blot,RT-PCR and immunohistochemistry were utilized to detect the HSP70 expression in the hippocampus and frontal lobes of rats after chronic mild stress.Results The sucrose favoritism in sucrose consumption test,the scores of crossing and the time for rats' retention in the center grid of open field test in long-term group were higher than those in short term group(P＜ 0.05).HSP70 expression in the hippocampus and frontal lobes of long-term group (0.81 ± 0.08,0.85 ± 0.08)detected by Western Blot and immunohistochemistry was higher that of short-term group (0.60 ± 0.06,0.85 ±0.07).HSP70 expression in the hippocampus of long-term group (0.90 ± 0.05,1.37 ± 0.38)detected by RT-PCR was higher that of short-term group(0.78 ± 0.04,1.08 ± 0.14) (P ＜ 0.05).Conclusion After long-term chronic mild stress,the depression-like behaviors decrease,and at the same time HSP70 mRNA and protein increase.

reression of M 2 receptor mRNA of rat myocardium intoxicated by soman under high altitude hypoxia. Methods The expressions of M 2 receptor mRNA under hypoxia, soman intoxication and soman intoxication under hypoxia were detected by semi quantitative RT PCR, respectively. Results The expression of M 2 receptor mRNA increased in the high altitude hypoxia group. Both simple soman intoxication and combined soman intoxication and hypoxia decreased the expression of M 2 receptor rapidly. But under hypoxia, the expression increased significantly at 12 h and 24 h. Conclusion M 2 receptor was sensitive to nerve agents. Compared to simple soman intoxication group, the expression of M 2 receptor increased in combined soman intoxication and hypoxia group. This may be one of the major factors leading to aggravation of the injury of heart function by nerve agents in high altitude area.

Objective To investigate the pathogenesis of avascular necrosis of the femoral head(ANFH) and to observe the effects of uhrashortwave diathermy treatment of ANFH through animal experiments.Methods A total of 30 New Zealand white rabbits were randomly divided into three groups:a control group,a model group,and a diathermy group.All the groups were injected with horse serum and methylprednisolone to induce ANFH.The path- ological effects were observed.Results The amount of osteoblast in the model group was significantly less than in the control group,while in the diathermy group it was significantly increased compared with the control group.The a- mount of osteoclast in the model and diathermy groups was significantly higher than in the control group,and in the diathermy group it was significantly more than in the model group.The thickness of femoral head cartilage in the mo- del and diathermy groups was reduced compared with the control group,while it was thicker in the model group than in the diathermy group.The empty cartilage cell lacunae ratios of the model and diathermy groups were significantly higher than for the control group,and the diathermy group showed significant degradation compared to the model group.The density of blood vessels under the cartilage in the model group was significantly less compared with the control group,while in the diathermy group it was significantly increased compared with the control group.The width of bone trabeculae in the model and diathermy groups was significantly less compared with the control group,while they were significantly wider in the diathermy group compared with the model group.The diameters of fat cells in the model and diathermy groups were increased compared with the control group,while they were significantly smaller in the dia- thermy group compared with the model group.The adipocyte area rates in the model and diathermy groups were signifi- cantly elevated compared with the control group,and rates in the model group were significantly elevated compared with the diathermy group.Conclusion Ultrashortwave diathermy is an effective treatment for early stage ANFH.

Aged ; Aged, 80 and over ; Atrial Fibrillation ; blood ; Biomarkers ; blood ; Female ; Humans ; Male ; Middle Aged ; Natriuretic Peptide, Brain ; blood

Objective To analyze the baseline clinical characteristics of the chronic kidney disease (CKD) in aged people in the clinic.Methods Patients aged 18 years or older in our CKD clinic from October 2003 to December 2012 were included in this study.According to their age patients were divided into 2 groups:aged CKD group:aged 65 or older and non-aged CKD group:younger than 65.A group of the elderly without CKD from health screening program were selected as aged non-CKD control group.Causes,distributions of stages and complications of CKD in three groups were analyzed.Results The major cause of the elderly CKD was hypertension,different from that of younger CKD.The distribution of CKD stage in the elderly was mainly in the G3b stage,different from that in the younger.Anemia and mineral bone disease presented in earlier CKD stage in the aged CKD patients and the prevalence was higher than in the aged non-CKD group.The prevalence of hypertension had no statistical difference between the two CKD groups,but hypertension control rate was lower in aged CKD patients.Conclusions The clinical characteristics including causes and renal stage are different between the young and aged CKD patients.Complications such as anemia and mineral bone disease presents in earlier renal stage in aged CKD patients which means we should monitor and interfere earlier.

Objective To compare the mRNA level of macrophage inflammatory protein-3α(MIP-3α) in 3 different mucosal epithelial cell lines. Methods RNA standards were prepared by in vitro transcription. One-step real-time RT-PCR was established and optimized using TaqMan EZ RT-PCR Core Reagents with TaqMan probes and primers specific to human MIP-3α mRNA sequence. The specificity of one-step real-time RT-PCR method was confirmed by sequencing the PCR products. The sensitivity and reproducibility of the method was examined by repeating the test 8 times with the same sample. Results The one-step real-time RT-PCR with a wide detection range is sensitive, reproducible. It was found that MIP-3α mRNA level in Caco-2 and T-84 cells was much higher than that in the HeLa cells. Conclusion High level of MIP-3α mRNA could be found in mucosal epithelial cells and difference in transcription level of MIP-3α may exist in epithelial cells from different mucosa.

To discover novel fluoroquinolone lead compounds as possible anti-infective or/and antitumor chemotherapies, combination principle of pharmacophore-based drug design, a series of novel tricyclic fluoroquinolone title compounds, [1,2,4]triazino[3,4-h][1,8]naphthyridine-8-one-7-carboxylic acid derivatives ( 5a-5p), were designed and synthesized with a fused [1,2,4]-triazine ring unit. Their structures were characterized by spectral data and elemental analysis and the in vitro antibacterial and anti-cell proliferation activities were also evaluated. The results showed that the titled compounds exhibited more significant inhibitory activities against drug-resistant bacteria (Methicillin-resistant Staphylococcus aureus and multi drug-resistant Escherichia coli strains) and three tested cancer cell lines (human hepatoma SMMC-7721, murine leukemia L1210 and human murine leukemia HL60 cells). Interestingly, SAR showed that compounds with electron-donating groups attached to benzene ring had stronger antibacterial activity than antitumor activity, but electron-withdrawing compounds displayed more potential antitumor activity than antibacterial activity, especially antitumor activity of nitro compounds was comparable to comparison doxorubicin. Thus, novel triazine-fused tricyclic fluoroquinolones as potent anti-infective or/and antitumor lead compounds are valuable to pay attention and for further development.
Animals ; Anti-Bacterial Agents ; chemical synthesis ; chemistry ; Antineoplastic Agents ; chemical synthesis ; chemistry ; Carboxylic Acids ; Carcinoma, Hepatocellular ; Cell Line ; Cell Proliferation ; Drug Design ; Escherichia coli ; drug effects ; Fluoroquinolones ; chemical synthesis ; chemistry ; HL-60 Cells ; Humans ; Leukemia L1210 ; Liver Neoplasms ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; Mice ; Naphthyridines ; Triazines

Objective:To observe the clinical effect of spinal manipulation on chronic, non-specific neck pain.Methods:Thirty patients with chronic, nonspecific neck pain were divided randomly into an observation group ( n=15) and a control group ( n=15). Patients in the observation group were given 20 minutes of a novel 4R spinal manipulation (resetting joint malalignment, resetting abnormal muscle, resetting joint stabiliazation, resetting sensorimotor control) twice a week for 2 weeks while the control group were given 20 minutes of medium frequency and high frequency conventional physiotherapy 4 times a week, also for 2 weeks. Before the treatment, right after, and one and three months later, both groups were evaluated using a visual analogue scale (VAS) and a neck disability index (NDI). Right before and after the treatment, cervical flexion and extension range of motion (ROM) were measured. The surface electromyography was employed to record the root mean square (RMS) of the EMG amplitude and the median frequency (MF) from the erector spinae and upper trapezius. Results:Before the treatment no significant differences were found in any of the measurements between the two groups. Afterward and one and three months later the average VAS, NDI and cervical ROM results of both groups had improved significantly, with the improvements in the observation group significantly greater than those in the control group on average. After 2 weeks of treatment, the average RMS and MF values had improved in both groups, again with the observation group′s average values significantly better than those of the control group.Conclusion:Spinal manipulation can effectively improve the strength and stamina of cervical muscle groups in patients with chronic, non-specific neck pain.

Objective To evaluate the effects of sub-acute alcoholism on cardiac structure and function, and investigate the mechanisms of aldehyde dehydrogenases-2 (ALDH2)alleviating the damage of heart caused by acute myocardial infarction.Methods The wild mice with ALDH2 (+/+) (WT group) and mice with knockout type ALDH2 (-/-) genotypes (KO group) were raised and then divided into four groups according to the presence or absence of sub-acute alcoholism: WT group (n=10), KO group (n=16), WT+alcoholism group(WT+E,n=10) and KO+alcoholism group(KO+E,n=10).The mice of WT+E group and KO+E group were fed with high-dose of ethanol(2 g/kg per day for 8 days), while the mice of WT group and KO group were treated with equal amount of saline instead.Acute myocardial infarction models were established in all mice after ethanol administration,and blood ethanol concentration, cardiac function, myocardial infarct size, the activity of ALDH2, and the key molecules of PI3K/Akt signal pathway and caspase-3 mRNA were detected one week after modeling.Statistical analysis was performed using SPSS 17.0.Differences in levels of detected biomarkers between groups were assessed using Chi-squared or One way ANOVA, and P<0.05 was considered to be statistically significant.Results (1) The mortality rates of WT group, KO group, WT+E group and KO+E group were 20.0%, 30.0%, 31.3% and 37.5%, respectively.(2)Compared with WT group and KO group, the blood ethanol concentration was higher and the damage of liver was more severe in WT+E group and KO+E group(P<0.05).(3)The fraction shortening of short axis of left ventricle(FS) and left ventricular ejection fraction were higher in WT group and WT+E group compared with KO group and KO+E group(P<0.05).(4) The area of myocardial infarction was largest in KO+E group, followed by KO group, WT+E group, and WT group (all P<0.05).(5) The activity of ALDH2 in WT group was higher than that in other groups, and the ALDH2 activity in KO+E group was lower than that in KO group (P<0.05).(6) There was no significant difference in expressions of PI3K among four groups.The level of p-Akt was highest in WT group, followed by WT+E group, KO group, and KO+E group (all P<0.05).The levels of caspase-3 mRNA was highest in KO+E group, followed by KO group, WT+E group, and WT group (all P<0.05).Conclusions Myocardial damage caused by acute myocardial infarction can be aggravated by sub-acute alcoholism, while ALDH2 protection can effectively alleviate the damage effects of sub-acute alcoholism on myocardial infarction.The mechanism of protective effects of ALDH2 on acute myocardial infarction may be related to attenuation of cardiocytes apoptosis mediated by PI3K/Akt signal pathway.