The paper introduces the background of YY/T 0841 standard, and interprets the main content of this standard. It is helpful to understand and applicate the standard.
Electrical Equipment and Supplies ; standards

Objective To explore the biological function of ICAM-1and LFA-1 in tumor metastasis,we observed the expression of ICAM-1(Intercellular Adhension Molecule-1)and LFA-1(Lymphocyte function related antigen-1)in cancer tissue and normal tissue.Methods A total of the samples were obtained from hospital,it includes 60 cases of breast cancer and 60 cases of normal mammary tissues.We observed the expression of ICAM-1 and LFA-1 by SP immunohistochemistry.We analyzed the relationship between the expression of ICAM-1,LFA-1and lymphatic vessel metastasis of breast cancer.Results The expression positive rate of ICAM-1and LFA-1 in breast cancer was obviously higher than that in normal mammary tissues(P

Objective: To explore therapeutic effect of levosimendan combined dobutamine on refractory heart failure (RHF).Methods: A total of 90 RHF patients, who were treated in our hospital from Apr 2014 to Oct 2015, were selected.According to random number table, patients were randomly and equally divided into dobutamine group and combined treatment group (received dobutamine combined levosimendan).Cardiac function improvement after treatment,and adverse reactions during treatment were compared between two groups.Results: After treatment, cardiac function improvement rate of combined treatment group was significantly higher than that of dobutamine group (86.7% vs.60.0%), P=0.02.Compared with before treatment, after treatment, there was significant reduction in dyspnea score, and significant rise in LVEF and stroke votume(SV) in both groups (P<0.05 or <0.01);compared with dobutamine group after treatment, there was significant reduction in dyspnea score [(1.58±0.78) scores vs.(1.02±0.47) scores], and significant rise in LVEF [(42.61±7.10)% vs.(50.21±7.82)%] and SV [(57.62±14.63)ml vs.(64.21±15.12)ml] in combined treatment group, P<0.05 or <0.01.Hypotension, myocardial ischemia and arrhythmia etc.occurred in both groups, but there was no significant difference in incidence rate of total adverse reactions (P>0.05).Conclusion: Levosimendan combined dobutamine possess significant therapeutic effect on refractory heart failure and safety is good, which is worth extending.

OBJECTIVE: To study the apoptosis/proliferation of Kölliker organ supporting cells and to understand the prompting apoptosis factors in vivo in the supporting cells in the Kölliker organ by changing the environment of the cultured supporting cells in the Kliker organ in vitro, via the separation, culture and purification of the supporting cells in the K6lliker organ. METHOD: A combinatorial approach of enzymatic digestion and mechanical separation was employed to isolate and culture in vitro pure Kölliker organ supporting cells. The purity was tested by flow cytometry assay. And K6lliker organ supporting cells were harvested to detect the rate and cycle of apoptosis by flow cytometry after Annexin V/PI staining, to test the cell growth curve by MTT assay, and to observe the differential expressions of the Bcl-2, Caspase-3, Caspase-8 and Caspase-9 through the Realtime PCR and Western blot. The calcium, potassium and glutamate concentrations in the culture medium of these cells in vitro were changed to detect the survival rate of cells by MTT assay. RESULT: The purity of K6lliker organ supporting cells by flow cytometry assay was 96. 56%. And these cells showed no significant difference in apoptosis, but an evident linear growth. The results of Realtime PCR and Western blot showed that the expression of Bcl-2, Caspase-3, Caspase-8 and Caspase-9 mRNA and protein in all different time points kept stable. Furthermore, the elevation of extracellular Ca2+ might contribute to decrease the cell viability of supporting cells. And K+ participated regulation of cell viability in a concentration-depending way. However, glutamate appeared to be a protective factor in high concentration. CONCLUSION There is no significant apoptosis in vitro of the supporting cells in the Kölliker organ of rats, showing a linear growth. The Ca2+ in high concentration might contribute to the apoptosis factor of these cells. However, the K+ and glutamate appear to be protective factors in high concentration.
Animals ; Animals, Newborn ; Apoptosis ; Caspase 3 ; Cell Cycle ; Cell Proliferation ; Cell Survival ; Cochlea ; cytology ; growth & development ; Flow Cytometry ; In Vitro Techniques ; Rats

Mitochondrial aldehyde dehydrogenase(ALDH2),one of the isoforms of aldehyde dehydrogenase,has multiple enzymatic functions including the activity of dehydrogenase and esterase.The metabolisms of ethanol,amino acids,biogenic amine,vitamin or steroid in the body produce various substances of aldehyde.With the help of co-factor NAD(P)+,ALDH2 can convert aldehydes into corresponding carboxylic acid,which plays a key role in reducing toxic effects of aldehydes on the body.It does not need co-factor when ALDH2 works as esterase.It can convert carboxylic ester or other acids into corresponding carboxylic acids or alcohols.Recently,it has been shown that the decrease of ALDH2 activity exacerbates multiple factors(such as ethanol,ischemia)-induced myocardial injury and accelerates the development of nitroglycerin tolerance.Therefore,the development of specific agonists of ALDH2 may provide a novel approach to the therapy and prevention of heart diseases.

BACKGROUND: Osteosarcoma cell OS-9607 is a cell line cultured from human osteosarcoma tissue. To construct cDNA library of this cell line so as to clone target gene from it.OBJECTIVE: To construct cDNA library of osteosarcoma cell OS-9607 by switching mechanism at 5' end of RNA transcript (SMART) technique and analyze its quality.DESIGN: Verified experimental study SETTING: Department of Orthopaedics, Changhai Hospital, Second Military Medical University of Chinese PLA MATERIALS: This experiment was carried out at the Department of Orthopaedics, Changhai Hospital, Second Military Medical University of Chinese PLA in May 2005. Osteosarcoma cell OS-9607 was cultured in CO2 incubator. Cells were pre-treated with DM for 30 minutes, recovered for 24 hours in standard culture medium. The whole RNA of osteosarcoma cell OS-9607 was isolated by RNAease reagent kit. The quantity and integrity of total RNA was detected by ultraviolet spectrometer and electrophoresis on a denaturing formaldehyde agarose gel stained by ethidium bromide (EtBr). METHODS: Total RNA was extracted from human osteosarcoma cell OS9607 and mRNA was isolated, cDNA of OS-9607 cells was acquired by in situ polymerase chain reaction (PCR) and long-distance PCR (LD-PCR),then the cDNA was ligated with dephosphorylated λ phage vector. The recombinants were constructed with SMART cDNA library construction kit.The size of cDNA inserts and the diversity of library were detected by PCR.MAIN OUTCOME MEASURES: The quality and integrity of total RNA, the titer of un-amplified and amplified library and recombination fraction; analysis of cDNA length of library RESULTS: ①The concentration of the isolated total RNA was determined and the measuring absorbance was between 260 nm and 280 nm. Its absorptance was about 1.9 A. 0.8％ denaturing formaldehyde agarose gel electrophorosis showed clear 28S rRNA and 18S rRNA,and the brightness rate of 28S rRNA and 18S rRNA (28S/18S) was 2:1. ②After linked to λ phage in vitro, packed and transfected host bacteria, the titer of unamplified library was 2.2×107 pfu/mL, and the titer of amplified 1 ibrary arrived to 4.5×1010 pfu/mL. The recombination fraction analysis showed that there were 8 blue plaques and 1 360 colorless plaques in one plate, so the recombination fraction was 99.5％. ③ The obtained cDNA library consisted of 1.6×106 recombinant bacteriophages and the average exogenous inserts of the recombinants was 1.5 kb.CONCLUSION: These results suggest that the obtained osteosarcoma cell OS-9607 cDNA library has high quality, which sets up a firm foundation for the further research on osteosarcoma cell OS-9607 and its elated genes screen.

Objective To study the effects of protein kinase C (PKC) inhibitor staurosporine (STS) on the proliferation and apoptosis of human cholangiocarcinoma QBC-939 cells and to explore its possible mechanism. Methods CCK-8 was used to detect the effects of PKC inhibitor STS on the proliferation of human cholangiocarcinoma QBC-939 cells. The effects of STS on the ultrastructural characteristics of QBC-939 cells were observed by routine transmission electron microscopy (TEM). The apoptosis rate and the cell cycle distribution of QBC-939 cells were detected by flow cytometry. The expression of cyclin B1,Cdk1 and p-Cdk1 in QBC-939 cells was detected by Western blotting. Results STS could significantly inhibit the proliferation of QBC-939 cells in a dose-dependent manner (P <0. 05) and the half inhibitory concentration ( IC50 ) of QBC-939 cells at 24th and 48th h was 334 nmol·L-1 and 118 nmol · L-1 , respectively. TEM observed that STS could induce typical apoptotic bodies and super-microstructural changes of QBC-939 cells. By Annexin V-FITC/ PI double labeling flow cytometry,we found that the apoptotic rate of QBC-939 cells after treatment with STS for 0,12,24 and 48 h was (10. 16±4. 52)% ,(22. 35±2. 19)% ,(34. 27±2. 30)% and (59. 70±5. 97)% ,respectively. By flow cytometry,compared with the control group,STS could significantly increase apoptosis rate of QBC-939 cells,decrease the percentage of cells in G0 / G1 phase and increase the percentage of cells in G2 / M phase (P<0. 05). Western blotting proved that the expression levels of cyclin B1 and Cdk1 proteins in the STS-treated QBC-939 cells were significantly decreased (P<0. 05),while the expression level of p-Cdk1 protein in the STS-treated QBC-939 cells was significantly increased ( P < 0. 05 ). Conclusion STS can significantly inhibit cell proliferation and induce apoptosis of human cholangiocarcinoma QBC-939 cells and the mechanism may be related to cell cycle arrest at G2 / M phase.

With the development of chemical analysis technology and data processing technology,metabonomics has become the important method in functional genomics study.In this paper,the definition and advangtages of metabonomics are provided.The implication of metabolomics in pharmacology are summarized and future perspectives are discussed.

Objective:To analysis the N-terminal amino acid sequence of 5D3Ag purified by mAb5D3 immunoaffinity column.Methods:SDS-PAGE and Western Blot were used to study the characteristics of 5D3Ag.From SDS-PAGE gel,5D3Ag was transferred to PVDF membrane and its N-terminal amino acid sequence was analysised with an auto amino acid sequencer.Results:SDS-PAGE of 5D3Ag showed a band on the gel with molecular weight of 43 kD.Western Blot study showed that mAb5D3 can recognize 5D3Ag.The N-terminal amino acid sequence was NQGSHGSEEQ.Conclusion:It is sure that 5D3Ag is unique to Osteosarcoma because of the purification method itself and the characteristics of mAb5D3.Compared with other reported Osteosarcoma associated antigen,5D3Ag is different from them in molecular weight and may be a novel Osteosarcoma associated antigen.With the help of N-teminal amino acid sequence,some primers may be designed to clone the whole length gene of 5D3Ag.The identification of 5D3Ag gene may be useful in understanding the biologic mechanics of Osteosarcoma and developing new diagnostic and therapeutic strategies.

Objective To investigate the differential expression of synaptophysin(SYN)in the developing rat cochlea,to analyze the relationship between the expression of SYN and auditory functional development,and to explore the source of the adenosine triphosphate(ATP)in the cochlea.Methods The immunofluorescence was applied to observe the differential expression of SYN in the cochlea of SD rats on different days(p1,p5,p10,p14 and p28)after parturition.Results No significant stain was observed in the Kolliker organ or the Corti's organ of P1,P5 and the top turn of the cochlea in P10.The SYN expression was turned on the outer spiral bundle,the inner spiral bundle,and the medial wall of the Deiters' cell of the cochlea in P14,P28 and the second or the third turn of Corti's organ of P10.All the spiral ganglion neurons(SGN)were stained in different aged cochlea.Conclusion The expression of SYN was different during development of rat cochlea.This difference was in favor of the configuration constructed between nerve endings and target cells.It also played a key role in the formation of correct auditory coding during auditory system development.The hair cells and the supporting cells,possible the source of ATP in the cochlea,contained ATP that maybe existed in a non-vesicular form or specific staining of non-SYN.