organizing science research papers(12)

8/8/2019 Organizing Science Research Papers(12)

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Organizing Science

Research Papers (12)

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(:)

Setting of research ():??

Research problem () :?

Quantitative specification of problem () :

Importance of problem () :,?


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(:)

Research objective ()

Methodology to achieve objective()

Anticipated results ()

Contribution to field ()


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( :) Cellular signal transduction includes many cell signalpathways, an area having received considerable research interest. Thecell endoplasmic reticulumERCa2+ pool attempts to maintain anion concentration balance of cellular calcium, which is an extremelyimportant organelle. Among the calcium, an ion is cellular ER toproceed with the protein translation, protein translocation, folding and

cellular ER translation protein to confirm ER Ca2+ pool plays asignificant role. However, these functions should mainly cause theimplementation of many resident ER Ca2+binding proteins.Theexistence of many Ca2+-binding proteins in the ER is widelydocumented. Additionally, although passive in producing ER inside thehigh concentration, these proteins have an important physiologicalfunction. Additionally, the protein kinase C accepts some physiologicalfunctions after stimulation. Protein kinase C belongs to serine-threonine kinase about the message transduction The cell-relatedhormone or growth factor is used to proceed to the next step foractivation of phospholipase C, thus producing DAG. Finally, proteinkinase C is activated.


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( :) However, in addition to having many isoforms, PKC candifferentiate between conventional PKCs (a, b1, b2, and g), novel PKCs(d, e,and), atypical PKCs (and/) . Previous research focused onER and how to regulate the calcium concentration of the intracellularpathway. . Therefore, these PKC isoforms activate to some degreecorrelation. However, PKC cannot clearly identify the different types of

cell lines that inhibit ER Ca2+ sequestering activity. PKC has also notbeen investigated with respect to the intracellular Ca2+ pool.

Still, ER Ca2+ pool has not been investigated withrespect to PKC isoforms in cell signal transduction, the functions ofPKC and DNA transcription or translation as well as variousintracellular pathways. Moreover, conventional cell culture methods

can not thoroughly understand the cellular pathway of PKC, makingthe cellular apoptosis mechanism unclear as well.

The inability to thoroughly understand protein kinaseC and the cell signal pathway will negatively impact the physiologicalcharacteristics of PKC with respect to cellular life and death.


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(:) Based on the above, we should attempt to

understand what role macrophage secretion cytokine plays

in the human immune system.

To do so, NO can be examined, as foundbetween different macrophage cells, RAW 264.7 and J774.

An identical stimulator and inhibitor can then be inserted

into RAW 264.7 and J774 murine macrophage cells,

allowing RAW264.7 and J774 cells not only to produce the

cytokine but also to induce apoptosis physiology.

Moreover, NO content and cell apoptosis can be examined.

Furthermore, RAW 264.7 and J774 cells can be treated

with the LPS in different glucose concentration media.


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(:) As anticipated, analysis results can indicate

that NO and other cytokines can locate many signal

pathways of the macrophage. Additionally, the NO content

and protein kinase C of cellular signal regulation can

identify the RAW 264.7 or J774 cell morphology in differentglucose concentration media.

Results in this study can demonstrate that, in

addition to possibly inducing the cell apoptosis pathway,

NO can promote the human immune system to achieve an

appropriate balance between cytokines.


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( :) Commonly found in the RAW 264.7 cell,

endoplasmic reticulum (ER) calcium pool plays a

significant role in regulating the concentration of cellular

calcium ion. Additionally, , ER calcium pool can facilitate

protein translation, protein transfer, and proteinembellishment. According to recent investigations,

elevated intracellular Ca2+ concentration, [Ca2+]i, can

initiate apoptosis; in addition, [Ca2+]i increases before

genome fragmentation and cell death. As well known, as amajor intracellular reservoir of Ca2+ in nonmuscular cells,

endoplasmic reticulum ER is essential for many cellular

functions, including protein processing within ER.


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( :) However, while previous studies investigated how murine

macrophage cell line regulates the signal pathway of ER calcium pool,

exactly how the signal pathways of cellular TNF-, NF-B, and MAPK

regulates the concentration of cellular calcium ion remains unclear.

For instance, cell ER pool investigations have notidentified the signal pathways of TNF-, NF-B, and MAPK within an

accuracy of 80, thus making it impossible to determine how RAW

264.7 cell regulates the signal pathway.

The inability to thoroughly understand the signal

pathways of intracellular TNF-, NF-B, and MAPK makes itimpossible to determine what role ER calicium pool and induced

cytokine play in the RAW 264.7 cell.


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(:) Based on the above, we should analyze TNF- and NF-B of

the RAW 264.7 cell by using cell culture methods, providing insight into

how the cell signal pathway and immunity regulation of nitro oxide in

cellular endoplasmic reticulum (ER) calcium poolare related. TNF-

,NF-B and MAPK can also be determined to be closely associated

with the signal pathways of both human diseases such as cancer.

To do so, the RAW 264.7 cell can be analyzed using

Griess reagent (1% Sulfanilamide, 0.1% NED) and chemiluminescence.

The PKC protein can then be analyzed using western blot analysis.

Next, NF-B and MAPK can be analyzed using primary and secondaryantibodies. Additionally, the mouse fibroblasts can be used by adding

L929 to serial dilutions of the conditioned media at 5104 cells per well

(in 96-well plates), followed by treatment with 1 g/ml actinomycin D.

Moreover, after 24 h of treatment, the viability of cells can be measured

by MTT assay. Finally, a standard curve can be defined using TNF-.


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(:) As anticipated, analysis results can indicate

that the TNF-,NF-B and MAPK can be found in the RAW

264.7 cell of the cellular endoplasmic reticulum (ER)

calcium pool signal pathway. Additionally, this pathway can

be understood with respect to elucidating thecharacteristics of cancer.

Results of this study can provide further insight

into not only the signal pathways of intracellular TNF-, NF-

B, and MAPK, but also the role in which ER calicium pool

and induced cytokine play in the RAW 264.7 cell.


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organizing science research papers(12)

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