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Cellular signal transduction includes many cell signal
pathways, an area having received considerable research interest. The
cell endoplasmic reticulum ER Ca2+ pool attempts to maintain an
ion concentration balance of cellular calcium, which is an extremely
important organelle. Among the calcium, an ion is cellular ER to
proceed with the protein translation, protein translocation, folding and
cellular ER translation protein to confirm ER Ca2+ pool plays a
significant role. However, these functions should mainly cause the
implementation of many resident ER Ca2+binding proteins. The
existence of many Ca2+-binding proteins in the ER is widely
documented. Additionally, although passive in producing ER inside the
high concentration, these proteins have an important physiological
function. Additionally, the protein kinase C accepts some physiological
functions after stimulation. Protein kinase C belongs to serine-
threonine kinase about the message transduction The cell-related
hormone or growth factor is used to proceed to the next step for
activation of phospholipase C, thus producing DAG. Finally, protein
kinase C is activated.
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However, in addition to having many isoforms, PKC can
differentiate between conventional PKCs (a, b1, b2, and g), novel PKCs
(d, e,and), atypical PKCs (and/) . Previous research focused on
ER and how to regulate the calcium concentration of the intracellular
pathway. . Therefore, these PKC isoforms activate to some degree
correlation. However, PKC cannot clearly identify the different types of
cell lines that inhibit ER Ca2+ sequestering activity. PKC has also not
been investigated with respect to the intracellular Ca2+ pool.
Still, ER Ca2+ pool has not been investigated with respect
to PKC isoforms in cell signal transduction, the functions of PKC and
DNA transcription or translation as well as various intracellular
pathways. Moreover, conventional cell culture methods can not
thoroughly understand the cellular pathway of PKC, making the cellular
apoptosis mechanism unclear as well. The
inability to thoroughly understand protein kinase C and the cell signal
pathway will negatively impact the physiological characteristics of PKC
with respect to cellular life and death. Therefore, exactly
what role macrophage secretion cytokine plays in the human immune
system must be investigated.
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Exactly what role macrophage secretion cytokine plays
in the human immune system can be investigated. To
do so, NO can be examined, as found between 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.
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
different glucose 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. (NOTE : Add 2-4
more sentences that describe more thoroughly how the proposed
method contributes to a particular field or sector)
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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 protein embellishment. 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 a major
intracellular reservoir of Ca2+ in nonmuscular cells, endoplasmic
reticulum ER is essential for many cellular functions, including
protein processing within ER. 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.
(NOTE : Add 2-4 sentences that describe characteristics of the
problem or statistics that reflect its severity)
()
For instance, cell ER pool investigations
have not identified 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
it impossible to determine what role ER calicium pool and
induced cytokine play in the RAW 264.7 cell.
Therefore, TNF- and NF-B of the RAW 264.7 cell must be
analyzed by using cell culture methods, subsequently 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 must also be
determined to be closely associated with the signal pathways of
both human diseases such as cancer.
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TNF- and NF-B of the RAW 264.7 cell can be analyzed by
using cell culture methods, subsequently 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
secondary antibodies. 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-. 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 the characteristics 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. (NOTE : Add 2-4 more
sentences that describe more thoroughly how the proposed method contributes
to a particular field or sector)
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