Science Project Summary (12)

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Our recent scientific effort
addressed how to understand the physiological characteristics of PKC
with respect to cellular life and death.
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.
(cont)
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.
(cont)
Therefore, we investigated what role macrophage
secretion cytokine plays in the human immune system.
NO was examined, as found between different
macrophage cells, RAW 264.7 and J774. An identical stimulator and
inhibitor were then 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 were examined. Furthermore,
RAW 264.7 and J774 cells were treated with the LPS in different
glucose concentration media. Analysis results
indicated 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 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)

Our group has
thoroughly explored the difficulty in elucidating the signal
pathways of intracellular TNF-, NF-B, and MAPK.
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.
(cont)
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 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, we analyzed 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.
(cont)
The RAW 264.7 cell was analyzed using
Griess reagent (1% Sulfanilamide, 0.1% NED) and chemiluminescence.
The PKC protein was then analyzed using western blot analysis. Next,
NF-B and MAPK were analyzed using primary and secondary
antibodies. Additionally, the mouse fibroblasts was 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 was measured by MTT assay.
Finally, a standard curve was defined using TNF-.
Analysis results indicated 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 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)
Further details can be found at
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