Objective To establish the model of amyotrophic lateral sclerosis (ALS) with selective motor neuron disorder by organotypic spinal cord cultures, and analyze the role of astrocyte in the pathagenisis of ALS.Methods Organotypic spinal cord cultures were prepared using lumbar spinal cord slices from 8-day-old SD rat pups. The threohydroxyaspartate (THA) was applied into culture medium to establish ALS organotypic spinal cord cultures model. Motor neurons survival was evaluated by monoclonal SMI-32 immunohistochemical staining and glial fibrillary acidic protein staining to show astrocyte survival.Results Compared with the control group, there was significantly astrogliosis in the anterior horn and surrounding white matter in THA 100 μmol/L group, and the level of gliosis was increased followed the elongation of THA interference time. With the increasing of the number of astrocyte, the morphology of astrocyte was changed.Conclusion There is significantly astrogliosis in the anterior horn and the time of astrogliosis is markedly earlier than the time of motor neuron loss in the ALS model intervented with THA.

Objective To analyze the correlations between pediatric intersive cave unit (PICU) nurses′job satisfaction and their perception of job characteristics. Method One hundred and thirty-six PICU nurses from a women and children′s hospital of Guangzhou participated in the study and the job diagnostic survey (JDS) and Minnesota satisfaction questionnaire short-form (MSQ20) were used to investigate the relationship between the job characteristics and job satisfaction. Results The average score on general satisfaction was (3.57 ± 0.41). The dimensions like skill variety, task integrity, feedback from job and feedback from others, had a significant positive impact on general satisfaction (all P<0.05). Conclusion Nurses generally have a relatively middle level of job satisfaction and measures should be taken to improve their job satisfaction as well as the quality of nursing.

Objective:To explore and analyze the points-selection rules in acupuncture treatment of mammary gland hyperplasia (MGH) by data mining and statistical method. Methods:Clinical literatures about the treatment of MGH with acupuncture published in the recent 16 years were retrieved from Chinese Journal Full-text Database (CJFD) and established into a database by Excel. The SPSS 20 version software and Clementine 12.0 version software were adopted to analyze the frequency and association rules of points-selection in the treatment of MGH with acupuncture. Results:The top 3 points used most frequently in acupuncture treatment of MGH were Danzhong (CV 17), Taichong (LR 3) and Zusanli (ST 36); points from the Stomach Meridian of Foot Yangming and Liver Meridian of Foot Jueyin were most commonly used; the commonly selected points were predominantly distributed in thoracic and abdominal regions and lower limbs; emphasis on the combination use of local and distal points; of the specific points, the five Shu-Transmitting points were mostly used; association analysis showed that the associations among Taichong (LR 3), Danzhong (CV 17) and Zusanli (ST 36) were the most significant. Conclusion: The data mining results substantially accord with the general rules of acupuncture-moxibustion theories in traditional Chinese medicine, able to reflect the points-selection principles and features in acupuncture treatment of MGH and provide evidence for the points selection in the treatment of MGH in acupuncture clinic.

Colony-Forming Units Assay ; Cryopreservation ; methods ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Immunotoxins ; immunology ; Leukocytes, Mononuclear ; cytology ; Ricin ; immunology ; T-Lymphocytes ; cytology ; immunology ; Time Factors

Teachers are the most important resources of colleges and univesities. Setting up the teachers' competence model of medical colleges and universities is the basis of guaranting the quality of medical education, which accords with the need of the post training of teachers and can be the basis for teachers' competence assessment.

The clinical analgesic effect of electro-acupuncture (EA) stimulation (EAS) on breakthrough pain induced by remifentanil in patients undergoing radical thoracic esophagectomy, and the mechanisms were assessed. Sixty patients (ASAIII) scheduled for elective radical esophagectomy were randomized into three groups: group A (control) receiving a general anesthesia only; group B (sham) given EA needles at PC4 (Ximen) and PC6 (Neiguan) but no stimulation; and group C (EAS) electrically given EAS of the ipsilateral PC4 and PC6 throughout the surgery. The EAS consisting of a disperse-dense wave with a low frequency of 2 Hz and a high frequency of 20 Hz, was performed 30 min prior to induction of general anesthesia and continued through the surgery. At the emergence, sufentanil infusion was given for postoperative analgesia with loading dose of 7.5 μg, followed by a continuous infusion of 2.25 μg/h. The patient self-administration of sufentanil was 0.75 μg with a lockout of 15 min as needed. Additional breakthrough pain was treated with dezocine (5 mg) intravenously at the patient's request. Blood samples were collected before (T1), 2 h (T2), 24 h (T3), and 48 h (T4) after operation to measure the plasma β-EP, PGE2, and 5-HT. The operative time, the total dose of sufentanil and the dose of self-administration, and the rescue doses of dezocine were recorded. Visual Analogue Scale (VAS) scores at 2, 12, 24 and 48 h postoperatively and the incidence of apnea and severe hypotension were recorded. The results showed that the gender, age, weight, operative time and remifentanil consumption were comparable among 3 groups. Patients in EAS group had the lowest VAS scores postoperatively among the three groups (P<0.05). The total dose of sufentanil was 115±6.0 μg in EAS group, significantly lower than that in control (134.3±5.9 μg) and sham (133.5±7.0 μg) groups. Similarly, the rescue dose of dezocine was the least in EAS group (P<0.05) among the three groups. Plasma β-EP levels in EAS group at T3 (176.90±45.73) and T4 (162.96±35.00 pg/mL) were significantly higher than those in control (132.33±36.75 and 128.79±41.24 pg/mL) and sham (136.56±45.80 and 129.85±36.14 pg/mL) groups, P<0.05 for all. EAS could decrease the release of PGE2. Plasma PGE2 levels in EAS group at T2 and T3 (41±5 and 40±5 pg/mL respectively) were significantly lower than those in control (64±5 and 62±7 pg/mL) and sham (66±6 and 62±6 pg/mL) groups. Plasma 5-HT levels in EAS group at T2 (133.66±40.85) and T3 (154.66±52.49 ng/mL) were significantly lower than those in control (168.33±56.94 and 225.28±82.03) and sham (164.54±47.53 and 217.74±76.45 ng/mL) groups. For intra-group comparison, plasma 5-HT and PGE2 levels in control and sham groups at T2 and T3, and β-EP in EAS group at T3 and T4 were significantly higher than those at T1 (P<0.05); PGE2 and 5-HT levels in EAS group showed no significant difference among the different time points (P>0.05). No apnea or severe hypotension was observed in any group. It was concluded that intraoperative ipsilateral EAS at PC4 and PC6 provides effective postoperative analgesia for patients undergoing radical esophagectomy with remifentanil anesthesia and significantly decrease requirement for parental narcotics. The underlying mechanism may be related to stimulation of the release of endogenous β-EP and inhibition of inflammatory mediators (5-HT and PGE2).

The chemical consitituents from cytotoxic fraction of the Callicarpa nudiflora extract were isolated and purified by a combination of HP-20 macroporous resin, silica gel and Sephadex LH-20 column chromatographies. The structures were elucidated on the basis of the spectroscopic data and comparison of their spectroscopic data with reported data. The cytotoxicity was evaluated by the MTT assay. The 50% and 70% EtOH elutions of EtOH-extract showed significant cytotoxic activities, leading to the isolation of twelve compounds, which were identified as luteoloside(1), lutedin-4'-O-β-D-glucoside(2), 6-hydroxyluteolin-7-O-β-glucoside(3), lutedin-7-O-neohesperidoside(4), rhoifolin (5), luteolin-7, 4'-di-O-glucoside (6), forsythoside B (7), acteoside (8), alyssonoside (9), catalpol(10), nudifloside(11), and leonuride(12). Compounds 3-6, 10 and 12 were isolated from this genus for the first time, and compound 9 was isolated from this plant for the first time. The cytotoxicity assay demonstrated that flavonoids 1-6, in various concentrations, showed monolithic proliferation inhibitory activities against Hela, A549 and MCF-7 cell lines. Compounds 3, 5 and iridoid glycoside 11 possessed higher cytotoxicacivities. In short, flavonoids are the main components of cytotoxic extract from C. nudiflora, while phenylethanoid glycosides are the predominant ingredient but inactive to cancer cell lines. In addition, the minor iridoid glycoside expressed weak cytotoxic activity.
Callicarpa ; chemistry ; Cell Proliferation ; drug effects ; Cytotoxins ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Humans ; MCF-7 Cells ; Molecular Structure

Objective To study the effect of injured podocytes on glomerular maturation and its underlying mechanism in neonatal mice.Methods Single i.p.injection with puromycin aminonucleoside (PA,0.1 mg/g BW) was given to ICR neonatal mice at day 1 after birth (1 dpp). Littermates injected with normal saline (NS) were used as control.Animals were examined for urine protein,blood pressure,kidney weight/body weight (KW/BW),renal histology at 2,4,8,12, 30,60 and 90 dpp (n=6～9 for each group).Immunohistochemistry and quantitative RT-PCR were performed to examine the expression of WT-1,CD31,VEGF,Flk-1,Ang-1,Ang-2,Tie-1 and Tie-2.Results Mice with PA injection had lower kidney weight and body weight at all time points as well as lower KW/BW at 4,8,12 dpp when compared with NS controls.Electron microscopy revealed nearly complete foot process effacement and segmental microvillous transformation as early as 1 day after PA injection.PA-injected kidneys showed fewer capillary loops and decreased maturation index as well as less CD31-positive endothelium in cortical glomeruli at 12 dpp. Glomerular mesangial injury and developing glomerulosclerosis along with proteinuria were noted in PA-injected kidneys starting from 30 dpp.Significantly increased systolic blood pressure was detected at 60 dpp in PA mice.Compared with NS injection,PA injection significantly induced decreased mRNA expression of Flk-1 and Tie-2 as well as increased expression of Ang-1,without obvious changes of VEGF at 2 dpp.Conclusions Podocytes in neonatal kidney of ICR mice are susceptible to PA. Such podocyte injury can alter the expression of VEGF and angiopoietin system in glomeruli,leading to abnormal development of glomerular capillaries,and subsequent proteinuria,hypertension and glomerulosclerosis.

Selection and affinities of Cd binding peptides were assayed by phage random dodecapeptide library and affinity chromatography. Two Cd binding peptides were obtained, it was found that the affinities of Cu~ 2+ ,Co~ 2+ ,Zn~ 2+ ,Ni~ 2+ for Cd binding peptides were higher than that of Cd~ 2+ and Cr~ 2+ after detection of the amplified Cd binding peptides displayed phages to different heavy metal-charged resins; the detoxification of E.coli to Ni~ 2+ and Cd~ 2+ was enhanced when infected by Cd binding peptide displayed phages as compared with those of the control. The interaction of Cd binding peptide displayed phages with heavy metals charged resins was also observed under microscope. The work would be of great value and consequences for the study of interaction between heavy metals and proteins(peptides), as well as thedetoxification and bioremediation of heavy metals.

Abstract: Objective To establish the duplex TaqMan RT-PCR method for detection of Entamoeba histolytica and Giardia lamblia in fecal samples. Methods Primer pairs and probes for Entamoeba histolytica and Giardia lamblia were designed and duplex TaqMan RT-PCR amplification system was constructed. PCR products were inserted into the pUC57 plasmid, and the lower limit of detection of the method was determined. Clinical stool samples were tested in order to evaluated the efficacy of the method. Results The detection limits of duplex TaqMan RT-PCR were 31.6 copies/μL for Entamoeba histolytica and 32 copies/μL for Giardia lamblia, respectively. Of the total of 212 clinical stool samples tested, all 3 samples with E. histolytica-positive patients by microscopy were positive by PCR, while 1 from 209 samples with E. histolytica-negative patients by microscopy were positive by PCR, and the remaining samples were negative. For Giardia lamblia, all 8 samples positive by microscopy were positive by PCR, and 1 from 204 sample with a microscopy-negative patient was positive by PCR, and the remaining samples were negative. The amplification product sequencing and blast analysis were used to confirm that the amplified sequence in the specimen of a patient with negative microscopy but positive PCR belongs to the targeted pathogen, supported by clinical symptoms and laboratory test results. PCR results for other diarrhea-causing pathogens were negative, indicating no cross-reactivity. Conclusions The dual TaqMan RT-PCR method developed in this study can not only detect microscopy-positive samples of Entamoeba histolytica and Giardia lamblia but also can detect samples that were missed by microscopy, with higher sensitivity than the microscopy method. Further, this detection method does not cross-react with other diarrhea-causing pathogens, including cross-react with other diarrhea-causing pathogens including Iodamoeba butschlii, Blastocystis hominis, Plesiomonas, Aeromonas, Salmonella, Shigella, Sphaerozoum fuscum, and Entamoeba hartmani, thus has a good specificity.