Background and purpose: High expression of telomerase and telomere stability are two common features in tumor cell. hTERT is a catalytic subunit of telomerase, TRF2 is extremely important to maintain the length and stability of telomerase. This study was to construct the recombinant adenovirus mediated shRNA to hTERT and TRF2, and to investigate the inhibitory effects of the vector by solo-inhibiting and connect-inhibiting in the MCF-7 cells, in order to present a new approach to the gene therapy for breast cancer. Methods: rAd-hTERT and rAd-TRF2 were constructed and the expression of hTERT mRNA and TRF2 mRNA were tested by FQ-PCR 48 hours after transfecting in MCF-7 cells. Apoptosis was observed by flow cytometry 1 to 6 days after transfection. Results: ①At 48 hours after transfection, the results of FQ-PCR showed that compared to PBS group, the expression of hTERT in rAd-hTERT group was obviously decreased and the inhibition ratio was about 86%, but TRF2 had not been obviously inhibited (P>0.05);the expression of TRF2 in rAd-TRF2 group was obviously decreased and the inhibition ratio was about 80%, but hTERT had not been obviously inhibited (P>0.05);in rAd-hTERT/rAd-TRF2 group, the inhibition ratio of hTERT and TRF2 were about 88% and 85%. Comparing rAd-hTERT/rAd-TRF2 group with rAd-hTERT group and rAd-TRF2 group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). Otherwise, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). ②The result of flow cytometry showed that apoptosis was induced at the first day after transfecting in rAd-hTERT group and rAd-TRF2 group, the most obvious apoptosis was in the 3rd to 5th days,at the peak in the 5th day, and decreased in the 6th day after transfection. The apoptosis ratio of rAd-hTERT group was 46.2%, rAd-TRF2 group was 43.5%. The apoptosis ratio of rAd-hTERT/rAd-TRF2 group was 46.2% at first day, 68.5% at the second day, the most obvious apoptosis was in the 3rd to 6th days and was 77.6% in the 6th days in rAd-hTERT/rAd-TRF2 group. There were significant differences in apoptosis ratio in solo-inhibiting and connect-inhibiting(P<0.05). In addition, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences in apoptosis ratio(P>0.05). Conclusion: ①Target sequence of RNAi which aimed at hTERT gene and TRF2 gene was designed efficiently, and the RNAi expression vectors were seen in vivo study efficiently and specifically inhibited the correspond gene expression and promoted cell apoptosis in MCF-7 cells. ②rAd-hTERT vector and rAd-TRF2 vector have no synergistical effect and antagoinstical effect on inhibiting hTERT gene and TRF2 gene mRNA expressing in MCF-7, but there was synergistical effect in terms of the induction of apoptosis. So association-RNAi-technique targeting to the genes of telomere length and stability can effectively promote tumor cell into apoptosis and inhibit breast cancer cell growth. RNAi technique of connecting correlation genes is a more effective gene therapy strategy.

Objective To investigate the expression of human epidermal growth factor receptor?2(HER?2)and Survivin protein in gastric cancer tissue,and explore its relationship with clinicopathological characteristics. Methods Immunohistochemistry assay was used for detection of HER?2 and Survivin protein expression in 70 gastric cancer biopsy samples. The relationship with the clinicoathological parameters in patients with gas?tric cancer was analyzed. Results The expression of HER?2 and Survivin protein was correlated with tumor differentiation ,TNM stage and lymph node metastasis(all P<0.05),but not with sex,age,the diameter of tumor. The correlation analysis of positive rate and the semi?quantitative pro?tein expression showed that the expression of Survivin and HER?2 was significantly positively related(all P<0.01). Conclusion The positive ex?pression of HER?2 and Survivin protein in gastric cancer correlates with tumor differentiation ,TNM stage and lymph node metastasis. HER?2 ex?pression significantly correlates with Survivin at the protein level in gastric cancer tissues.

We evaluated an astaxanthin overproducing Phaffia rhodozyma JMU-MVP14, and developed astaxanthin high-yielding fermentation process. We analyzed several fermentation parameters, i.e., biomass, astaxanthin and total carotenoids content to compare the characteristics of P rhodozyma JMU-MVP14 and the original strain through flask fermentation experiments. We conducted batch and fed-batch fermentation experiments in 7 L fermentor to investigate the effects of pH controlling models and feeding medium compositions on the production of astaxanthin. We further evaluated the capability and practical value of P rhodozyma JMU-MVP14 by fed-batch cultivation in the 1 m3 fermentor. Flask fermentation experiments revealed that P. rhodozyma JMU-MVP14 produced high yield of astaxanthin and carotenoids with specific productivity of astaxanthin and specific productivity of total carotenoids of 6.01 mg/g and 10.38 mg/g. Results of batch culture experiments in the 7 L fermentor showed that controlling the pH by ammonia auto-feeding was better than discontinuously adjusting pH value at 6.0 with regard to the high productivities of biomasses and astaxanthin. This P. rhodozyma strain synthesized astaxanthin partially linked to the growth with the Ks and pmax of 0.20 h ' and 21.73 g/L, respectively. Results of batch-fed fermentations in 7 L fermentor indicated that the complex feeding medium consisted of 50% glucose, 0.5% yeast extract and 0.3% corn steep syrup had lower astaxanthin productivity than the simple feeding medium containing only 50% glucose, which produced biomass, volumetric productivity of astaxanthin, volumetric productivity of total carotenoids, specific productivity of astaxanthin and total carotenoids at 32.81 g/L, 155.99 mg/L, 4.94 mg/g, 399.99 mg/L and 12.19 mg/g, respectively. As fed-batch cultured in 1 m3 fermentor, P rhodozyma JMU-MVP14 yielded 85.11 g/L of biomass, 279.96 mg/L of volumetric productivity of astaxanthin, 618.01 mg/L of volumetric productivity of total carotenoids, 3.29 mg/g of specific productivity of astaxanthin and 7.26 mg/g of specific productivity of total carotenoids. Additionally, P rhodozyma JMU-MVP14 cell contained 21.54% of protein, 41.34% of carbohydrate and 34.31% of lipid. These comprehensive results suggest that P. rhodozyma JMU-MVPl14 has great practical prosperity related to its strong ability to produce astaxanthin and good value byproducts.
Basidiomycota ; genetics ; growth & development ; metabolism ; Batch Cell Culture Techniques ; Carotenoids ; biosynthesis ; Culture Media ; Fermentation ; Hydrogen-Ion Concentration ; Industrial Microbiology ; Kinetics ; Xanthophylls ; biosynthesis

To establish a rat model of rheumatoid arthritis (RA) with kidney deficiency syndrome.

Objective: To construct a female TCM management platform that integrates pulse diagnosis information to facilitate women's health status identification and conditioning of menstrual cycle related health problems. Methods:Using cloud computing technology, taking the EHR and knowledge base as the core, designing the TCM health information collection questionnaire according to the characteristics of women.s menstrual cycle related health problems, and incorporating the data collected by the Chinese medicine pulse diagnosis bracelet, the knowledge base can assist the development of a health management plan, designing a platform framework for combining the PC side of the doctor with the patient mobile APP. Results: The platform was conducive to the realization of more objective TCM consultation and pulse diagnosis information collection and long-term continuous monitoring, and can assist in the identification of women's menstrual problems and the development of health conditioning plans. Conclusion: The platform has good operability and practicability for the implementation of TCM health management for women with menstrual problems.

Objective: To explore pulse characteristic of the health female college students and their relationship with menstrual cycle. Methods: The subjects were 17 female college students of Jiangxi University of Traditional Chinese Medicine. Pulse test was performed with pulse bracelet. The detection time was every day in the wake and has not yet get up and wake up at any time before going to bed with a pulse-taking bracelet, and recorded the current menstrual cycle information. The pulses of the wake that has not yet get up and wake up at any time before going to bed took consistency analysis of the record with Kruskal Wallis Test. The pulse of menstruation stage analysis was compared with chi-square test. Results: The pulses of the wake that has not yet get up and wake up at any time before going to bed consistency was poor except rapid pulse (P＞ 0.05) . The occurrence frequency of slippery pulse and string-like pulse were 55.21％ and26. 03％ respectively. The pulses of premenstrual period, post menstrual period and menstrual period were not much different from others. The occurrence frequency of moderate pulse, string-like pulse, unsmooth pulse, slippery pulse in intermenstrual period respectively were 40.68％, 44.07％, 30.51％, and 40.68％. The occurrence frequency of moderate pulse, string-like pulse, unsmooth pulse, slippery pulse in-non intermenstrual period respectively were 17.38％, 24.69％, 10.45％, 56.30％. The difference was significant (P ＜ 0.05) . Conclusion: The female college students are mainly in terms of slippery pulse. The occurrence frequency slippery pulse in intermenstrual period respectively are less common. The string-like pulse, moderate pulse, unsmooth pulse in intermenstrual period are more common than in-non intermenstrual period.

Objective To study the correlation of quantitative electroencephalography (qEEG) parameters with outcomes of adults who sustained coma after cardio-pulmonary resuscitation.Methods The clinical data of coma patients after cardio-pulmonary resuscitation,admitted to our hospital from March 2008 to August 2014 were retrospectively analyzed.Glasgow coma scale (GCS) was performed.EEG was registered and recorded at least once within 7 d of coma after cardio-pulmonary resuscitation:electrodes were placed according to the international 10-20 system,using a 16-channel lay out.The qEEG parameters defined as burst suppression ratio (BSR),brain symmetry index (BSI),[delta+theta]/[alpha+beta] ratio (DTABR) were analyzed at the same time.Follow-up was performed 3 months after onset and the outcomes of these patients were assessed by Glasgow outcome scale (GOS).The correlations of GOS scores with qEEG parameters were analyzed.Results Sixty patients with GCS scores-＜ 8 after cardio-pulmonary resuscitation were included.BSR1 and BSR2 were negatively correlated with GOS scores 3 months after onset (r=-0.723,P=0.000;r=-0.651,P=0.000);DTABR was positively correlated with GOS scores 3 months after onset (r=0.270,P=0.037).BSI and DTABR were not correlated with GCS scores.BSR1 and BSR2 were negatively correlated with GCS scores (r=-0.562,P=0.000;r=-0.429,P=0.001).Conclusion The qEEG parameters are correlated with outcomes after cardio-pulmonary resuscitation.

OBJECTIVES: This study aimed to investigate the effect of new biomimetic micro/nano surfaces on the osteoclastic differentiation of RAW264.7 macrophages by simulating natural osteons for the design of concentric circular structures and modifying graphene oxide (GO). METHODS: The groups were divided into smooth titanium surface group (SS), concentric microgrooved titanium surface group (CMS), and microgroove modified with GO group (GO-CMS). The physicochemical properties of the material surfaces were studied using scanning electron microscopy (SEM), contact-angle measurement, atomic force microscopy, X-ray photoelectron spectroscopy analysis, and Raman spectroscopy. The effect of the modified material surface on the cell biological behavior of RAW264.7 was investigated by cell-activity assay, SEM, and laser confocal microscopy. The effect on the osteoclastic differentiation of macrophages was investiga-ted by tartrate-resistant acid phosphatase (TRAP) immunofluorescence staining and quantitative real-time polymerase chain reaction (qRT-PCR) experiments. RESULTS: Macrophages were arranged in concentric circles along the microgrooves, and after modification with GO, the oxygen-containing groups on the surface of the material increased and hydrophilicity increased. Osteoclasts in the GO-CMS group were small in size and number and had the lowest TRAP expression. Although it promoted the proliferation of macrophages in the GO-CMS group, the expression of osteoclastic differentiation-related genes was lower than that in the SS group, and the difference was statistically significant (P<0.05). CONCLUSIONS Concentric circular microgrooves restricted the fusion of osteoclasts and the formation of sealing zones. Osteomimetic concentric microgrooves modified with GO inhibited the osteoclastic differentiation of RAW 264.7 macrophages.
Graphite/pharmacology* ; Titanium/pharmacology* ; Haversian System ; Macrophages ; Cell Differentiation ; Oxides/pharmacology* ; Surface Properties

BACKGROUNDThe brain death confirmation tests occupy a different position in each country's diagnostic criteria (or guideline); the choices of tests are also different. China brain death criteria include clinical judgment and confirmation tests. This study aimed to confirm the preferred confirmatory test and complementary confirmatory tests.

METHODSWe did a clinical brain death determination on deep coma patients, and then divided them into brain death group and non-brain death group. According to the Chinese standards for determining brain death, both the groups accepted confirmatory tests including electroencephalograph (EEG), somatosensory evoked potentials (SEP), and transcranial Doppler (TCD). The sensitivity, specificity, false positive rate, and false negative rate were calculated to evaluate the accuracy of the confirmatory tests.

RESULTSAmong the 131 cases of patients, 103 patients met the clinical criteria of brain death. Respiratory arrest provocation test was performed on 44 cases and 32 cases (73%) successfully completed and confirmed that they have no spontaneous breathing. Of the three confirmation tests, EEG had the highest completion rate (98%) and good sensitivity (83%) and specificity (97%); TCD had followed completion rate (54%) and not good sensitivity (73%) and specificity (75%); SEP had the lowest completion rate (49%), good sensitivity (100%), and not good specificity (78%). After the combination of SEP or TCD with EEG, the specificity can increase to 100%.

CONCLUSIONSThe completion rate of respiratory arrest provocation test remains a problem in the clinical diagnosis of brain death. If the test cannot be completed, whether to increase a confirmatory test is debatable. SEP had an ideal sensitivity, and the specificity will reach 100% after combining with TCD or EEG. When a confirmed test was uncertain, we suggest increasing another confirmatory test.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Brain Death ; diagnosis ; physiopathology ; Evoked Potentials, Somatosensory ; physiology ; Female ; Humans ; Male ; Middle Aged ; Ultrasonography, Doppler, Transcranial ; Young Adult