Objective To explore the influencing factors for the purity and viability of primary cultured cortical neurons of rat,and optimize the separating and cultivating conditions of the cortical neurons.Methods The primary cotical neurons were cultured in a serum-free culture system of B27-supplemented neurobasal medium.The differences in purity and viability of primary cultured neurons between embr-yonic rat group and newborn rat [postnatal 24 h and 5 d] group were evaluated by morphology,immunocytochemistry of neuron-specific enolase(NSE) and trypan blue staining.The changes of neurons purity and viability in different trypsin digestion time(0 min,5 min and 15 min) at different environment temperatures(20 ℃ and 30 ℃) were assessed by immunocytochemistry and trypan blue staining.Results The primary cultured neurons from fetal and newborn rats grew well.There was no significant difference in embryonic rat and postnatal 1 d newborn rat group[(91.30?1.03)%,(89.50?1.78)% respectively in purity;and(98.20?0.58)%,(97.10?0.98)% respectively in viability].The neurons from 5-days newborn rat were inferior to that from fetal and 1-day newborn rat in purity and viability[(82.00?1.25)% and(92.87?1.56)% respectively].Shortening operation time and adjusting digestion time according to the environment temperatures could improve neuronal viability:A digestion for 15min at the environment temperature of 20 ℃ or for 5 min at 30 ℃ could acquired cells with higher viability [(98.20?0.58)% and(96.70?0.64)% respectively].Conclusions It is an easy,practical choice to culture priamry cortical neurons from postnatal 1 d newborn rats.Optimizing the separating and cultivating condition(environment temperatures,digest time,et al.) will improve the neurons purity and viability.

Objective To assess the reliability and validity of the effect evaluation index system for health education of coronary heart disease.Methods A total of 306 patients with coronary heart disease in four third-grade and A-level hospitals extacted by convenient sampling method were surveyed.Project analysis,exploratory factor analysis,correlation analysis and so on were applied to analyze and select items.Then the Cronbach's alpha coefficient,test-retest reliability,and content validity of the scale were tested.Then the original system was modified and the eventual system was formed.Results The scale was consisted of 8 first-dimensions and 35 indexes.The Cronbach's α of the total system was 0.832,and the α of subscales were from 0.629 to 0.882.The test-retest reliability of the total system was 0.966,and the testretest reliability of subscales were from 0.848 to 0.942.The S-CVI/Ave of system was 0.99,and the S-CVI/UA of indexes was 0.91.The Pearson correlation coefficient of the indexes with their dimensions were from 0.574 to 0.916,which were higher than the coefficient of the indexes with other dimensions.Factor analysis supported the eight-factor structure of the system,which could explain 65.61％ of the total variance and the factor loading of each index were from 0.430 to 0.938.Conclusions The system is reliable and valid,and can provide a unified reference standard for effect evaluation of health education of coronary heart disease.

Objective To establish a scientific,standard and comprehensive effect evaluation index system for health education of coronary heart disease.Methods Based on KABP (knowledge,attitude,belief,practice,KABP) model of health education,the effect evaluation index system for health education of coronary heart disease was formulated by documentary analysis,qualitative research,questionnaire survey,Delphi method,and analytic hierarchy process.Results The authoritative coefficient,determination coefficient and familiarity coefficient was 0.89,0.91,0.86,respectively.The coordination coefficients of the first-dimension and the second-dimension were 0.295 and 0.302,respectively.The index system consisted of 6 first-dimensions,45 second-dimensions.The weight coefficient of health knowledge,health belief,health behavior,physical signs,quality of life,and satisfaction degree of health education was 0.153 8,0.153 8,0.366 1,0.086 3,0.153 8,0.086 3,respectively.Conclusions The effect evaluation index system for health education of coronary heart disease is reliable and valid,which can provide a quantitative basis for effect evaluation of health education.

Objective To evaluate cerebrospinal fluid adenosine deaminase(CSF-ADA)activity in the diagnosis of tuberculous meningitis(TMB), and to observe its dynamic changes. Methods A total of 160 patients were included and were divided into two groups: 76 cases of TBM and 84 cases of non-TBM.Among the cases of non-TBM, there were 36 cases of bacterial meningitis, 30 cases of viral meningitis and 18 cases of cryptocoocal meningitis. All the patients were measured with their CSF-ADA activity by Enzymecoupled assay(Trinder method)and 47 patients of TBM were measured again after 2 weeks' and 6 weeks'antitubercular therapy. Results were expressed as(-x)± s. Mann-Whitney U test and paired-samples t test were used. Results CSF-ADA activity in TBM group was(12.9 ±6.4)U/L, while that in the non-TBM group was(6.0 ± 4.1)U/L, the difference was of statistical significance(U = 7.860, P ＜ 0.05). With the cutoff value of 9 U/L, the sensitivity and specificity to differentiate TBM from non-TBM was 84.21% and 83. 33%, respectively. CSF-ADA activity decreased in TBM patients after antitubercular treatment.Conclusions CSF-ADA activity can be an effective laboratory marker for early differential diagnosis of TMB with the cut-off value of 9 U/L. Dynamic changes of CSF-ADA activity may be a indicator for the effect of antitubercular treatment.

Objective To evaluate the influence of two different education approaches on diabetic knowledge of type Ⅱ diabetes.Methods 79 type Ⅱ diabetes cases were collected from March 2010 to May 2010,and divided into experimental group and control group according to admission time.Experimental group was those in the first-half,and received figure dialogue education.Control group was those in the last-half,and received traditional heath education.Diabetes knowledge assessment questionnaire was used to evaluate knowledge level before and after education.Results Knowledge level was higher after education than that before education in both groups.No differences was found between the two groups ( P ＞ 0.05 ).All subjects in experimental group thought figure education could improve the study interesting,68.3％ subjects thought mastering knowledge about diabetes was useful,and 80.0％ subjects expect discussion by group.However,74.4％ subjects in control group thought traditional education could improve the study interesting,15.4％ subjects thought mastering knowledge about diabetes was useful,and 46.2％ subjects expect discussion by group.Conclusions Compare with traditional education,figure education although could not increase the knowledge level about diabetes significantly,but could highly increase the interesting of study.

The tail suspension test (TST), forced swimming test (FST) and chronic unpredictable mild stress (CUMS) model were used to evaluate the anti-depressant effect of supercritical CO2 extract from Compound Chaigui Fang (FFCGF). A nuclear magnetic resonance (NMR)-based metabonomics combined with multivariate statistical analysis was performed to explore the mechanism of FFCGF. Rats were conducted by CUMS procedure for 28 days and drugs were administrated at the same time. The body weight, sucrose preference, crossings and rearings in open-field tests were evaluated and the urine was collected simultaneously. The metabonomic profiles of rats' urine were analyzed by NMR and potential biomarkers were searched by multivariate statistical analysis. The results showed that administration of FFCGF significantly decreasing the immobility time in FST and TST and improving rats' body weight, sucrose preference, crossings and rearings in CUMS, which were indication that the anti-depressant effect of FFCGF was abvious. Significant differences in the metabolic profile of the CUMS treated group and the control group were observed, which were consistent with the results of behavioral tests. Decreased levels of acetic acid, succinic acid, 2-oxidation glutaric acid and citric acid and increased glycine and pyruvic acid in urine were significantly affected by the CUMS procedure and the 6 biomarkers were reversed evidently after administration of FFCGF. These changes were suggestion that the anti-depressant mechanism of FFCGF was associated with energy metabolism, lipid metabolism and amino acid metabolism.
Animals ; Antidepressive Agents ; chemistry ; isolation & purification ; pharmacology ; therapeutic use ; Behavior, Animal ; drug effects ; Body Weight ; Carbon Dioxide ; chemistry ; Depression ; drug therapy ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; therapeutic use ; Male ; Mice

OBJECTIVETo compare anatomical differences between subinguinal and ilioinguinal approaches for the treatment of acetabular fracture and investigate clinical therapeutic effect of subinguinal approach.

METHODSSeven fresh human bodies were chosen, comparative study were performed on the right and left side on the same specimen. Ilioinguinal approaches were adopted on the left and subinguinal were adopted on the right. Inner part of incision started to sun wild above pubic symphysis at 2 cm, and lateral incision ranged from iliac to anterior superior spine about 5 cm. Length and transverse diameter of the first window exposed and lliopsoas freeness were tested and compared. Fifteen acetabular fracture patients treated through subinguinal approach were compared from May 2010 to August 2012. Among all patients, including 12 males and 3 females aged from 20 to 65 years old with an average of 40.6 years old. Matta criteria were used to evaluate clinical outcomes.

RESULTSLength and transverse diameter of the first window exposed and lliopsoas freeness through subinguinal approach were better than through ilioinguinal approach (P<0.01). In 15 patients with acetabular fracture, 10 patients obtained anatomical reduction and 10 patients got satisfied reduction in accordance with Matta criteria. X-ray results of all patients were excellent.

CONCLUSIONCompared with ilioinguinal approach, subinguinal approach could enlarge visualization of the first window and simplify surgical procedure. It is an ideal approach to expose anterior and anterior-medialis wall of acetabulum and anterior hip capsule.

Acetabulum ; injuries ; surgery ; Adult ; Aged ; Female ; Fracture Fixation ; methods ; Fractures, Bone ; surgery ; Groin ; Humans ; Male ; Middle Aged

Acupuncture Points ; Acupuncture Therapy ; Adult ; Female ; Humans ; Male ; Middle Aged ; Pharyngitis ; therapy ; Young Adult

Mounting evidence has shown that side population (SP) cells are enriched for cancer stem cells (CSCs) responsible for cancer malignancy. In this study, SP technology was used to isolate a small subpopulation of SP cells in human gallbladder cancer cell line GBC-SD, and SP cells which had superior potential for proliferation in vitro and tumorigenesis in vivo were identified. Importantly, the abundance of GBC-SD SP cells was increased by a transforming growth factor-β (TGF-β)-induced epithelial-mesenchymal transition (EMT), and this effect was accompanied with a strong up-regulation of ABCG2 mRNA expression, and a decreased sensitivity to mitoxantrone. SP cells were restored upon the removal of TGF-β and the reversion of the cells to an epithelial phenotype, and smad3-specific siRNA reduced SP abundance in response to TGF-β. In conclusion, TGF-β-induced EMT by smad-dependent signaling pathway promotes cancer development and anti-cancer drug resistant phenotype by augmenting the abundance of GBC-SD SP cells, and a better understanding of mechanisms involved in TGF-β-induced EMT may provide a novel strategy for preventing cancer progression.

Objective: To investigate the effect of acupuncture plus mild hypothermia on neurological function impairment score, cerebral infarct size and apoptosis-related factors in cerebral ischemia reperfusion injury (CIRI) rats.Methods: Sixty healthy male Sprague-Dawley (SD) rats were routinely reared for 1 week. Ten rats were randomly selected as the sham operation group and 10 rats as the blank control group, while the remaining 40 rats were subjected to preparing the middle cerebral artery occlusion (MCAO) model by modified filament occlusion method. The 40 MCAO rats were further randomly divided into a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, with 10 rats in each group. Rats in the sham operation group, the blank control group and the model group did not accept treatment except binding; rats in the acupuncture group received acupuncture treatment; rats in the mild hypothermia group received mild hypothermia treatment; rats in the acupuncture plus mild hypothermia group received acupuncture and mild hypothermia treatment. 72 h after the treatment, neurological function impairment score was performed; the infarct area ratio was determined by 2,3,5-tripheyl tetrazolium chloride (TTC) staining; apoptosis of brain cells was observed by TUNEL method; the expressions of Bcl-2, Bax and Caspase-3 were detected by immunohistochemistry.Results: Compared with the blank control group and the sham operation group, the neurological function impairment score, cerebral infarct area ratio, apoptosis, and the expressions of Bax and Caspase-3 in the model group were significantly increased, while the expression of Bcl-2 was significantly decreased, and there were significant between-group differences (allP<0.05). After the treatment, there were statistically significant differences among the treatment groups in the neurological function impairment score, cerebral infarct area ratio and apoptosis in the ischemic side of rats, as well as the expressions of Bcl-2, Bax and Caspase-3 (allP<0.05), and from the figures, tables and statistical analysis, it was found that a better tendency in the acupuncture plus mild hypothermia group than the acupuncture group or mild hypothermia group.Conclusion: Acupuncture plus mild hypothermia can protect the brain cells by improving neurological function impairment, decreasing cerebral infarct area ratio, reducing the number of apoptotic cells in the ischemic area and regulating the expressions of apoptosis related proteins to inhibit apoptosis.