Biomedical subjects

Rong Liang

Publications and source records attributed to Rong Liang.

At least 19 recordsLinked to original sources

PubMed2006

Localization of cystathionine beta synthase in mice ovaries and its expression profile during follicular development.

BACKGROUND: In vitro fertilization (IVF) researches have suggested that cystathionine beta synthase (CBS) is involved in oocyte development. However, little is known about the regional and cellular expression patterns of CBS in the ovary. The purpose of this study was to analyze the localization of CBS in mice ovaries and to investigate the expression profile during follicular development. METHODS: We used in situ hybridization and immunohistochemical analysis to determine CBS expression in the ovaries of female Balb/c mice. Then the follicles were collected from F1 (C57BL x Balb/c) mice and cultured in vitro. With the method of semi-quantitative RT-PCR, we also investigated the expression profile of CBS during follicular development. RESULTS: CBS was absent in the oocytes, although it was ubiquitously expressed in the ovary with the strongest expression in follicular cells at all stages. In late antral follicles, CBS expression was markedly higher in granulosa cells located close to the antrum and in cumulus cells around the oocyte. The semi-quantitative RT-PCR showed that CBS mRNA was detected in follicles at all stages in vitro. In cumulus-oocyte complexes superovulated, CBS expression also increased rapidly. CONCLUSIONS: CBS was located mainly in the follicular cells in the ovaries. The level of CBS expression is high in follicles during folliculogenesis in mice. Differences in the CBS expression profile between oocyte and follicular cells suggest a role for CBS as a mediator in interactions between oocyte and granulosa cells.

Animals↗

PubMed2006

[Cloning, identification, and cellular localization of a down-regulated gene fragment related with Down Syndrome].

OBJECTIVE: To clone a novel gene and explore its expression patterns in tissues and cells, so as to find its role in the process of encephalopathy in DS. METHODS: On the base of our previous microarray's result together with the tissue type, we chose EST AI480014 to carry out RACE, then analyzed its expression profiles in liver, spleen, kidney, heart, brain by multi-tissues Northern blot, after that semi-quantitive RT-PCR was used to reexamine the expression profiles. Furthermore, we used ISH to find whether aim gene expressed in neuroglial cells cultured in vitro. Finally we performed semi-quantitive RT-PCR to explore whether it expressed differently between DS and normal. RESULTS: We gained a 682 bp new cDNA fragment (DQ275636) which expressed in all the tissues examined and had no alternative splices in them. It expressed highly in brain especially in frontal lobe and hippocampus. According to the ISH result we convinced that it expressed in neuroglial cells. Using bioinformatics we mapped DQ275636 to chromosome 5q14. CONCLUSION: We have obtained a new gene fragment based on the above results. According to its expression character and tissue type, it can be suggested that this gene has a probable role in the process of encephalopathy in DS.

Base Sequence↗

PubMed2006

Glucosamine sulfate-induced apoptosis in chronic myelogenous leukemia K562 cells is associated with translocation of cathepsin D and downregulation of Bcl-xL.

Cathepsin D (cat D) reportedly plays an important role in certain apoptotic processes, the downstream pathways of which involve release of cytochrome c (cyt c) from mitochondria and activation of the caspase cascade. Previous studies revealed that the B-cell lymphoma 2 (Bcl-2) family members Bax or Bid play important roles in apoptotic signal transduction between cat D and mitochondria. Here, we show that glucosamine sulfate (GS) inhibits the proliferation and induces apoptosis of human chronic myelogenous leukemia K562 cells in vitro. GS interfered with the maturation of cat D. Activation of caspase-3, cleavage of poly-(ADP-ribose)-polymerase, release of cyt c, and downregulation of Bcl-xL accompanied GS-induced apoptosis, and these processes were inhibited by the cat D inhibitor pepstatin A. However, we did not detect any altered gene expression of Bcl-2, Bax, or Bid during apoptosis. Translocation of cat D from the lysosome to the cytosol was observed in GS-treated K562 cells. These findings suggest that GS-induced K562 cell apoptosis involves the translocation of cat D from the lysosome to the cytosol. Furthermore, our findings suggest that downregulation of Bcl-xL (but not Bcl-2, Bax, or Bid) connects cat D and the mitochondrial pathway, which causes the release of cyt c and activation of the caspase cascade during GS-induced apoptosis of K562 cells.

Apoptosis↗

PubMed2006

Priming with DNA encoding E2 and boosting with E2 protein formulated with CpG oligodeoxynucleotides induces strong immune responses and protection from Bovine viral diarrhea virus in cattle.

The objective of this study was to develop an optimal vaccination strategy for Bovine viral diarrhea virus (BVDV). The E2 protein of BVDV plays a major protective role against BVDV infection. In order to be able to compare DNA, protein and DNA prime-protein boost regimens, a plasmid was constructed encoding a secreted form of the NADL strain E2 protein (pMASIA-tPAsDeltaE2). Furthermore, a pure secreted recombinant DeltaE2 (rDeltaE2) protein was produced. The rDeltaE2 protein was formulated with a combination of Emulsigen and CpG oligodeoxynucleotide. Groups of calves were immunized with pMASIA-tPAsDeltaE2 or with rDeltaE2, or first with pMASIA-tPAsDeltaE2 and then with rDeltaE2. To evaluate the protection against BVDV, calves were challenged with BVDV strain NY-1 after the last immunization. Although all immunized calves developed humoral and cellular immune responses, the antibody responses in the DNA prime-protein boost group were stronger than those elicited by either the DNA vaccine or the protein vaccine. In particular, E2-specific antibody titres were enhanced significantly after boosting the DeltaE2 DNA-primed calves with rDeltaE2 protein. Moreover, protection against BVDV challenge was obtained in the calves treated with the DNA prime-protein boost vaccination regimen, as shown by a significant reduction in weight loss, viral excretion and lymphopenia, compared with the unvaccinated calves and the animals immunized with the DNA or protein only. These results demonstrate the advantage of a DNA prime-protein boost vaccination approach in an outbred species.

Animals↗

PubMed2006

[Rac1 expression and its effects on the cell cycle progression and apoptosis in human acute leukemic cell line HL-60].

The study was aimed to investigate the expression of Rac1 in human acute leukemic cell line HL-60 and effect of Rac1 on cell cycle progression and apoptosis. The mRNA expression of Rac1 in HL-60 cell line and normal human peripheral blood mononuclear cells (PBMNC) were examined by semi-quantitative RT-PCR. After transfection of HL-60 cells with different concentrations of Rac1 antisense oligodeoxynucleotide (ASODN) by means of FuGENE6, the survival, cell cycle, apoptosis of HL-60 cells were observed through MTT assay, FCM test, Wright-Giemsa, acridine orange/ethidium bromide (AO/EB) and Annexin V-FITC/PI staining test respectively. The results showed that Rac1 relative amount in HL-60 was 0.84 +/- 0.13, while it in the normal PBMNC was 0.26 +/- 0.1 (P < 0.01); the expression of Rac1 in HL-60 cells was significantly upregulated. Compared with sense oligodeoxynucleotide (SODN), HL-60 cell viability, after exposure to ASODN at a concentration of 2.0 g/L decreased, (73.7 +/- 5.0)% vs (93.2 +/- 3.0)% (P < 0.01), while the proportion of G(1) cells increased as (52.1 +/- 6.8)% vs (31.6 +/- 4.7)% (P < 0.05), the percentage of Annexin V positive cells increased, (19.2 +/- 2.1)% vs (4.1 +/- 1.7)% (P < 0.01), and HL-60 cells were observed to have formation of apoptotic bodies. The data presented indicate that Rac1 may be involved in regulation of HL-60 cell cycle and apoptosis, promote overproliferation of HL-60 cells and inhibit their apoptosis.

Apoptosis↗

PubMed2006

[Inhibition of proliferation and induction of apoptosis and differentiation of leukemic cell line HL-60 by sodium valproate].

To investigate the influence of sodium valproate (VPA) on proliferation, apoptosis and differentiation of HL-60 cell line, effect of VPA in various concentrations on proliferation of HL-60 cells was detected by MMT; Wright-Giemsa staining was performed to observe the morphologic changes of HL-60 cells; NBT experiment was used to test the differentiation of HL-60 cells; flow cytometry was used to observe cell cycles and analyze the apoptosis. The results indicated that the changes of the growth curve showed inhibition of proliferation of HL-60 cells. After a 24-48 hours culture with 2 mmol/L VPA, the cells exhibited nuclear shrinkage, pyknosis fragmentation and appearance of apoptosis bodies. The percentage of the annexin V(+)/PI(-) cells which were apoptotic increased from 2.9% to 17.1%; hypodiploid peak was observed; the percentage of HL-60 cells in G(1) phase increased from 51.1% to 84.6% and the cells in S phase decreased from 37.9% to 14.4%. After a week culture with 0.25 mmol/L VPA, the cells exhibited characteristics of differentiation. The percentage of NBT positive cells was (47 +/- 2)%. It is concluded that VPA can inhibit the proliferation of HL-60 cells while inducing differentiation and apoptosis of these cells. The mechanism needs to be further studied.

Apoptosis↗

PubMed2006

[Difference of gene expression profiles between HL-60/VCR and HL-60 cells detected by human genome genechip].

This study was aimed to detect the gene expression profile changes between human acute leukemia cell line HL-60 and VCR-resistance HL-60, and to investigate the underlying mechanisms of MDR by using genechip technology. In experiments, mRNA were harvested using TrizoL reagent from these two cell lines, through RT-PCR, the biotinylated nucleotide were incorporated into the cRNA during the in vitro transcription reaction. The high quality RNA was hybridized to the gene expression array--human genome U133A developed by Affymetrix. It was scanned by G2500A GeneArray Scanner and the acquired image was analysed by a series of softwares. The results showed that 5,507 genes were differentially expressed between human acute leukemia cell line HL-60 and VCR-resistant HL-60. Compared with HL-60, 3,100 genes were up-regulated and 2,407 genes were down-regulated in VCR-resistant cell line. These genes were involved in different cell activities such as growth regulation and signal transduction. Among the genes with remarkable differential expression between the two cell lines, 435 were up-regulated and 605 were down-regulated. It is concluded that many different kinds of genes are involved in the mechanism of MDR and there is an intricate molecular network that controls the sensitivity of leukemia cells to the chemotherapeutic agents. Genechip is an efficient tool for parallel gene expression analysis.

Drug Resistance, Neoplasm↗

PubMed2005

[Preliminary exploration of the influence factors of degenerate oligonucleotide primered PCR of genome DNA].

OBJECTIVE: To explore the influence factors of the degenerate oligonucleotide primered PCR(DOP-PCR). METHODS: Genome DNA template from the mouse single oocyte or liver tissue were used to perform DOP-PCR. DOP-PCR was carried out with templates of different origin, different gradient dilution, with or without low melting point gel purified to wipe off the small fragment that might interfere with the following analysis, and then PCR of gene FTCD and CBS were carried out to evaluate the influence of these factors on the amplification efficiency and specificity. RESULTS: Compared with genome DNA template from mouse liver, the template from single oocyte had the same efficiency and specificity but a minor yield and different gradient dilution of DNA template had no effect on the efficiency and specificity. Furthermore, there was a higher specificity in the low melting point gel-purified DOP-PCR product than in untreated ones. CONCLUSION: We have got a satisfactory result and increased specificity from DOP-PCR product purified with the low melting point gel. Single oocyte of mice could be used for further investigation of special genes detection by DOP-PCR and of an optimization in the yield of the products.

Animals↗

PubMed2005

Immunization with plasmid DNA encoding a truncated, secreted form of the bovine viral diarrhea virus E2 protein elicits strong humoral and cellular immune responses.

The major protective antigen of bovine viral diarrhea virus (BVDV), the E2 protein, is cell-associated and not expressed on the cell surface. In this study we evaluated a DNA vaccine encoding various secreted versions of E2. In vitro analysis demonstrated that deletion of the transmembrane anchor and addition of the signal sequence of bovine herpesvirus-1 (BHV-1) (gDsDeltaE2) resulted in efficient secretion of E2 into the culture medium. In contrast, full-length E2, either without or with gDs (gDsE2), as well as truncated E2 without gDs (DeltaE2), remained entirely cell-associated. Mice immunized with plasmid encoding gDsDeltaE2 developed significantly higher IgG and virus neutralizing antibody titres compared to animals vaccinated with plasmid encoding E2, DeltaE2 or gDsE2. To optimize secretion of E2, the efficiency of gDs was compared with that of the tissue plasminogen activator signal (tPAs) sequence. In addition, the effect of the plasmid backbone was assessed by comparing two vectors. Four plasmids, pMASIA-gDsDeltaE2, pMASIA-tPAsDeltaE2, pSLKIA-gDsDeltaE2 and pSLKIA-tPAsDeltaE2, were constructed and administered intradermally to mice. The mice immunized with pMASIA-tPAsDeltaE2 developed the strongest and most balanced immune responses. Vaccination of cattle confirmed that pMASIA-tPAsDeltaE2 elicited both strong humoral and cellular immune responses and thus could be a candidate DNA vaccine against BVDV.

Animals↗

PubMed2005

Ectopic cervical anaplastic ependymoma.

Ependymomas generally arise in the central nervous system (CNS), although rare primary extraneural ependymomas have been observed. Reported herein for the first time is the case of a patient with primary ectopic cervical anaplastic ependymoma. The tumor was found in the right neck root region of a 35-year-old man. No additional tumor was found in the CNS or in other parts of the body. The patient received surgery and post-surgical local radiotherapy. Microscopically, the tumor consisted of round to oval cells with fine chromatin, distinct nucleoli, moderate nuclear atypia and numerous mitoses (>25/10 high-power fields) in a densely cellular growth pattern with characteristic fibrillary cytoplasm and formation of perivascular pseudorosettes. By immunohistochemistry, the tumor cells were positive for glial fibrillary acidic protein, epithelial membrane antigen (EMA), vimentin and S-100 protein. EMA staining showed a membranous as well as a paranuclear pattern of immunoreactivity. Electron microscopic studies revealed that tumor cells form micro rosettes, into which microvilli and cilia projected. The diagnosis was World Health Organization grade III anaplastic ependymoma. There is no evidence of local tumor recurrence or distant metastasis after 30 months follow up. The present case adds yet another unique example to the already diverse spectrum of head and neck neoplasms encountered in surgical pathology.

Adult↗

PubMed2005

[Expression of bFGF and VEGF in acute leukemia and its effects on HL-60 cell growth].

The study was to investigate the expression levels of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) in the serum of patients with acute leukemia and supernatants of leukemia cell lines as well as effects of VEGF-specific antisense oligodeoxynucleotides (ASODN) on the growth of HL-60 cells. Meanwhile the methods to evaluate the VEGF level in the serum of patients with acute leukemia were explored. The levels of bFGF and VEGF in the serum from 32 patients with acute leukemia and 10 healthy subjects and in the supernatants of 5 various human leukemia cell lines were quantified by means of the enzyme-linked immunosorbent assay (ELISA) and were compared. VEGF levels were evaluated not only without standardization but also after standardized by platelet and finally expressed as VEGF/PLT (pg/10(6)). After with different concentrations of VEGF ASODN, HL-60 cell viability was examined with MTT assay and VEGF levels in supernatants were measured with ELISA, respectively. The results showed that bFGF was detected (3 pg/ml) in 14 out of 32 serum samples from patients with acute leukemia, and the positive (37.5%) was significantly higher than that in healthy controls (10%) (P < 0.01). 3 out of 5 supernanant samples obtained from leukemia cell lines demonstrated positive for bFGF as well. There is no difference of the serum VEGF levels between leukemia patients and healthy controls, but the serum VEGF levels in the serum from leukemia patients were significantly higher than those in healthy controls (P < 0.05) after standardization. 4 out of 5 leukemia cell (U937 excluded) were found to express VEGF in the supernanant. After exposure of HL-60 cells to VEGF ASODN at a concentration of 0.5, 1 and 5 micromol/L for 24 hours, the cell viability gradually dropped down to lower levels (P < 0.05 vs controls). After treatment of HL-60 cells with VEGF ASODN at a concentration of 1, 5 and 20 micromol/L for 24 hours, the VEGF levels in supernatants of target cells decreased (P < 0.05 vs controls). The patients with acute leukemia represented the higher levels of serum bFGF and VEGF than controls. Most of leukemia cell lines expressed bFGF and VEGF at different levels. It is concluded that bFGF and VEGF both have effects on regulations of angiogenesis in acute leukemia, but VEGF plays a pivotal role. VEGF-specific ASODN may have a role in them VEGF expression downregulated. Different results may be obtained in the evaluation of VEGF levels in the serum of patients with acute leukemia if different calculation methods are used. The methods reported can measure leukemia associated VEGF more accurately.

Acute Disease↗

PubMed2005

[Effect of bone marrow stromal cells on the apoptotic sensitivity of HL-60 and HL-60/VCR cells].

This study was aimed to investigate the effects of human bone marrow fibroblastoid stromal cell line (HFCL) on chemosensitivity of acute myeloid leukemia sensitive HL-60 cell line and multidrug-resistant (MDR) HL-60/VCR cell line in vitro co-culture. Setting up co-culture system of HL-60 or HL-60/VCR cells in direct contact with HFCL cells, or with HFCL cells separated by transwell, and exposing HL-60 or HL-60/VCR cells to different concentrations of topotecon (TPT), morphologic evidence for apoptosis was determined by staining with Wright-Giemsa stain and acridine orange/ethidium bromide (AO/EB). Cell cycle, sub-G(1) and annexin V FITC staining were detected by flow cytometry. The expression of active caspase-3, Bcl-2 and Pgp was detected by Western blot. The results showed that HL-60 or HL-60/VCR cells treated by TPT revealed characteristic apoptotic morphological changes by Wright-Giemsa and AO/EB staining. The percentage of annexin V-positive cells and apoptotic cells decreased when they were cocultured with HFCL cells. The proportion of G(0)/G(1) HL-60 or HL-60/VCR cells treated by TPT increased and the sub-G(1) appeared significantly, but apoptotic and sub-G cells reduced after direct contact with HFCL cells. Meanwhile, although HL-60 or HL-60/VCR cells treated by TPT expressed activated caspase-3, and the expression of Bcl-2 decreased, the expression of activated caspase-3 decreased and Bcl-2 increased after direct contact with HFCL cells. In conclusion, HFCL stromal cells can prevent TPT-induced apoptosis in HL-60 and HL-60/VCR cells via modulation of Bcl-2 and active caspase-3.

Antineoplastic Agents↗

PubMed2005

[Role of Bcl-xL in the cathepsin D-associated apoptosis of K562 cells].

The purpose of study was to explore the possible functions of Bcl-xL in the glucosamine sulfate-induced apoptosis of chronic myeloid leukemia K562 cells. Light microscopy and Wright-Giemsa staining were used to investigate the morphologic evidences for apoptosis of K562 cells induced by glucosamine sulfate (GS); immunofluorescence was used to observe the translocation of cathepsin D and cytochrome C during the apoptosis; Western blot was performed to detect the expression of Bcl-xL, Bid, Bax in K562 cells treated by GS. The results showed that many vacuoles were observed in the cytoplasma of the K562 cells treated by GS; fluorescent signals of cathepsin D and cytochrome were fransformed from granules to disperse form by using immunofluorescence; the expression of Bcl-xL was found down-regulated in K562 cells treated by GS, but not in the cells pre-treated with pepstatin A; the significant changes were not detected in expression of Bax and Bid protein before or after apoptosis. It is concluded that Bcl-xL protein may mediate relationship between cathepsin D and mitochondia pathway, Cathepsin D may play an important role in the GS inducing apoptosis of K562 cells through downregulation of Bcl-xL expression.

Apoptosis↗

PubMed2005

[Effects of HCMV on phenotypes of parotid duct epithelial cells and its mechanisms].

OBJECTIVE: To investigate the effects of HCMV infection on phenotypes of parotid duct epithelial cells and relative mechanisms. METHODS: The expressions of immediate early antigen of HCMV, pan cytokeratin and cathepsin D etc. were detected by immunohistochemical staining in tissues of parotid cytomegalic inclusion disease. RESULTS: Cytokeratin which acts as an epithelial marker became negative while staining of Cathepsin D was intensified in parotid duct epithelial cells after infected by HCMV. CONCLUSION: It demonstrated that cytokeratin was lost through over-expression of Cathepsin D in parotid duct epithelial cells infected by HCMV.

Animals↗

PubMed2004

[The expression profiles of human chromosome 21 orthologues in mouse M II oocyte].

OBJECTIVE: To analyse the expression profile of orthologues of human chromosome 21 (HC21) in mouse M II oocytes, and to discuss the relationship between this expression profile and early embryonic development and further to find the possible reasons of DS phenotypes genesis. METHODS: cDNA array and Global RT-PCR methods were used to analyse and identify the expression profile of 93 HC21 orthologues in mouse M II oocytes. RESULTS: 26 of 93 orthologues were proved to be expressed in mouse M II oocytes and these genes were involved in many biological procedure including transcriptional, metabolism, ionic channel, and ubiquitin pathway etc. CONCLUSION: To our knowledge, this is first report about the HC21 orthologues expression profile in M II oocytes and this expression profile indicates that 26 of 93 HC21 orthologues may directly play important role in early development, and the biological information obtained from this experiment would be beneficial to understand imbalance of HC21 genes expression and the molecular mechanism of DS phenotypes genesis.

Animals↗

PubMed2004

[Effects of normal human bone marrow fibroblastoid stromal cell line on the proliferation of multiple myeloma cell line RPMI8226].

To investigate the effects of normal human bone marrow fibroblastoid stromal cell lines HFCL on the proliferation of multiple myeloma cell lines RPMI8226, the co-culture system of RPMI8226 with HFCL was established, the adhesion ratio was determined by MTT colorimetric assay, growth curves were determined by trypan blue exclusion, the mitotic index (MI) of RPMI8226 cell was observed by Wright-Giemsa staining. Flow cytometry and Western blot were used to study the changes of cell cycle and expression of proliferating cell nuclear antigen (PCNA), respectively. The results showed that in co-culture the adhesion ratio of RPMI8226 after 1 hour was 29.4%; the proliferation of RPMI8226 cell line in direct contact with HFCL cell line was inhibited as compared with RPMI8226 in suspension. No obvious changes were observed in RPMI8226 cell separated by transwell inserts. The percentage of G(1) phase cells of RPMI8226 cell line in direct contact with HFCL was higher than that of RPMI8226 in suspension, and the percentage of S phase cells was lower. Moreover, the MI of RPMI8226 cell line in suspension was higher than that of RPMI8226 cell line in direct contact with HFCL cell. The expression of PCNA in RPMI8226 cell line in suspension was higher than that of RPMI8226 cell in direct contact with HFCL cell. It is concluded that the normal human bone marrow fibroblastoid stromal cell HFCL inhibits the proliferation and progression of cell cycle of multiple myeloma cell line, RPMI8226.

Bone Marrow Cells↗

PubMed2003

[Investigation on circulation of Ao shi shang han jin jing lu (Ao's Golden mirror of cold pathogenic diseases) in Japan].

Being the earliest extant work on tongue diagnostics, Ao shi jin jing lu was block-printed and published in 1654 after it was spread into Japan, on the basis of which many kinds of manuscripts were made. This book not only exerted profound influence on the Kampo diagnostics in Edo age, but also laid down a foundation for the formation of pulse diagnostic school of tongue diagnostics in Kampo medicine.

Books↗

PubMed2003

[Effects of human fibroblastoid stromal cell line on proliferation of HL-60 cells and expression of VEGF].

To investigate the effects of normal human bone m arrow fibroblastoid stromal cell line (HFCL) on the proliferation of acute myeloid leukemia cell line HL-60 and expression of vascular endothelial growth factor (VEGF), establishing coculture system of leukemia cell line HL-60 and HFCL, growth data was obtained by cell counting. Mitotic index (MI) was observed under Wright-Giemsa staining. Flow cytometry and Western blot were used as assays for cell cycle and expression of proliferating cell nuclear antigen (PCNA) separately. VE GF levels were evaluated by using commercial ELISA kits. The results showed that compared with HL-60 cells without HFCL cells, the proliferation of HL-60 cells in direct contact with HFCL cells and with HFCL cells separated by transwell was inhibited. The MI of HL-60 cells without HFCL cells was highest followed by HL-60 cells separated by transwell and HL-60 cells in direct contact with HFCL cells. The expression of PCNA in HL-60 cells with HFCL cells were lower than HL-60 cells without HFCL cells. Meanwhile, the percentage of HL-60 cells in G1 phase cocultured with HFCL cells was higher than that without HFCL cells while the percentage of Sphase cells was lower. The levels of VEGF in HL-60 cells with HFCL cells were lower than that in HL-60 cells alone. In conclusion, the normal bone marrow fibroblastoid stromal cells inhibited the proliferation of HL-60 cells as well as the expression of VEGF.

Cell Cycle↗