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At least 955 records · Page 53Linked to original sources

PubMed2004

[Rapid serum-free culture of dendritic cells from human peripheral blood monocytes and their intracellular signal transduction].

OBJECTIVE: To explore the methods for rapid in vitro culture of the dendritic cells (DCs) from human peripheral blood monocytes (PBMCs) under serum-free conditions and ascertain whether intracellular signal transduction pathway differs between calcium ionophore (CI) and tumor necrosis factor (TNF)- alpha during their induction of dendritic cell differentiation. METHODS: PBMCs isolated from healthy donors were plated in serum-free medium supplemented with 50 ng/ml rhGM-CSF. Cells cultured overnight were induced to differentiate with 100 ng/ml A23187 or 50 ng/ml TNF-alpha, given before or 30 min after pre-treatment with 0.5 mug/ml cyclosporine A (CsA). After culture for 40 h, the cell morphology was observed under phase-contrast microscope, and the surface markers on treated PBMCs were analyzed by flow cytometry. MTT colorimetry was employed to assess the proliferation of the allogeneic T cells. RESULTS: PBMCs of healthy donors treated with 50 ng/ml rhGM-CSF in combination with 100 ng/ml CI or 50 ng/ml TNF-alpha for 40 h exhibited typical morphology of DCs with rapidly decreased CD14 expression and increased expressions of CD83 and co-stimulatory molecules (CD80 and CD86), showing also enhanced ability of stimulating allogeneic T cell proliferation. Calcineurin antagonist CsA inhibited the differentiation induced by CI, but not that induced by TNF-alpha. CONCLUSIONS: Under serum-free conditions, both CI and TNF-alpha are capable of inducing rapid DC differentiation from human PBMCs, but the intracellular signal transduction of CI-induced differentiation is different from that induced by TNF-alpha.

Calcimycin↗

PubMed2004

[Study on acute pancreatitis-associated lung injury induced by L-arginine in mice].

OBJECTIVE: To establish the model of acute pancreatitis (AP)-associated lung injury induced by L-arginine and study the role of TNF-alpha and ICAM-1 in the model mice. METHODS: For the establishment of the AP model, the mice were given the first injection (2 g/kg) of L-arginine intraperitoneally and then the second injection in the same way an hour later. At 12 h after the second injection, the animals' serum amylase activities were assayed by colorimetry, and the tumor necrosis factor-alpha (TNF-alpha) contents of their pancreatic tissue were determined by radioimmunoassay. At 24 h after the second injection, the histopathological changes in the animals' pancreatic and pulmonary tissues were observed. The pulmonary expression and cellular localization of intercellular adhesion molecule-1 (ICAM-1) were characterized by immunohistochemistry. RESULTS: The serum amylase activities, TNF-alpha content and ICAM-1 expression in lung tissue significantly increased in the AP group as compared with those in the control group. Histopathological examinations revealed that the acinar architecture was partially destroyed with necrosis, interstitial edema and inflammatory infiltration at 24 h after the second injection. CONCLUSION: TNF-alpha and ICAM-1 may play important roles in the mice with acute pancreatitis-associated lung injury.

Acute Disease↗

PubMed2004

[Experimental study on the penetrability of trypan blue to the rat prostate].

OBJECTIVE: To understand the penetrability of trypan blue to the normal prostate as well as to the inflammatory prostate and the prostate with benign hyperplasia in rats. METHODS: Sixty SD male rats were randomized into 4 groups: NP (normal prostate) group (n = 15), BP (bacterial prostatitis) group (n = 15), BPH (benign prostatic hyperplasia) group (n = 15), and BPH-BP (benign prostatic hyperplasia with bacterial prostatitis) group (n = 15). Five rats were taken from each group as non-staining controls (NC, n = 5 x 4) and the other 10 were injected by tail intravenation with 1% trypan blue and then the prostates were isolated from the rats killed by anaesthesia after 2 hours. The color of the prostates and other tissues of the animals were observed and the contents of the trypan blue in the tissues of the prostates were determined separately by colorimetry. RESULTS: Apart from the tissues of brains and spinal cord the surface and the inner tissues of the prostates with NP, BP, BPH and BPH-BP from the rats injected with the dye were also dyed blue similar to that of the muscles, livers, intestines and others. The content of the trypan blue in the tissues of the prostates with NP, BP and BPH-BP was obviously higher than those with NP and BPH. CONCLUSION: The penetrability of trypan blue with properties of ionization and larger molecular weight is high in either the normal prostate or the prostate with BP, BPH and BPH-BP, and much higher in inflammatory prostate than in the normal prostate and the prostate with BPH.

Animals↗

PubMed2003

[An experimental study on the effect of allitridin on inhibiting the expression of HCMV immediate-early antigens in vitro].

OBJECTIVE: To investigate the prophylactic, blocking and therapeutic effects of Allitridin on inhibiting HCMV proliferation by measuring the expression level of HCMV IEA in vitro and explore the mechanism of Allitridin anti-HCMVactivity. METHODS: The cytotocity of Allitridin was evaluated through MTT colorimetry and cell morphology. HCMV IEA levels were quantitatively detected by Flow Cytometry respectively under the following conditions: Allitridin was given before (pretreated for 24 h), during, or after viral inoculation in which serial doses (maximum tolerant concentration, MTC for human embryo lung cells, HEL) of Allitridin was used to treat HCMV infected HLE cells for different durations (24, 48, 72, 96 h) after viral infection. RESULT: The MTC of Allitridin was 9.60 mg x L(-1). Allitridin remarkably inhibited the expression of HCMV IEA in vitro. Within MTC, the inhibitory rate had a significant correlation with its dosage (r = 0.96). At the time of IEA highest expression (72 h after infection), inhibitory effect was the greatest (inhibitory rate: 89.3%). With pretreatment of Allitridin, the inhibitory rate was 28.6%. When Allitridin was used together with HCMV inoculation, IEA inhibitory rate was only 10.3%. CONCLUSION: Allitridin can inhibit HCMV, IEA expression in vitro remarkably which is probably one of the major mechanisms of Allitridin anti-HCMV activity because IEAs are the very important regulatory factors for the expression of all HCMV genes. Its therapeutic effect is the best at the peak stage of IE1 gene expression (72 h after infection) but it has low prophylactic and little blocking effect.

Allyl Compounds↗

PubMed2005

[Construction of the fusion gene of Hsp65 of Mycobacterium tuberculosis and EGFP and preparation of dendritic cell vaccine against tuberculosis].

AIM: To construct the fusion gene of Hsp65 of Mycobacterium tuberculosis H37Rv and enhanced green fluorescence protein (EGFP) and prepare dendritic cell (DC) vaccine. METHODS: Hsp65 DNA amplified by PCR was cloned into eukaryotic expression vector EGFP-C1. The recombinant plasmid pEGHsp65 was subsequently transfected into Hela cells, and the transfection rate was determined under confocal laser scanning fluorescence microscope at different times. RT-PCR was used to detect the expression of Hsp65 mRNA in Hela cells. The GM-CSF and IL-4 induced DCs from mouse bone marrow cells were transfected with recombinant plasmid pEGHsp65. Proliferation of unprimed splenocytes activated by transfected DCs was detected by MTT colorimetry. RESULTS: Restrictive enzyme digestion analysis (EcoR I, Bgl II) confirmed that Hsp65 DNA had been inserted into the vector pEGFP-C1. The recombinant plasmid pEGHsp65 was transfected into Hela cells and the expression of the fusion gene reached peak at the 48 hours after transfection. Expression of Hsp65 mRNA was detected in Hela cells by RT-PCR. DCs transfected with pEGHsp65 could stimulate the proliferation of unprimed splenocytes. CONCLUSION: The pEGHsp65 fusion gene was successfully constructed and DCs transfected with the pEGHsp65 might be a candidate vaccine for tuberculosis.

Animals↗

PubMed2005

[The role of MAPK signal transduction pathway in killing tumor cells by Mtb-Ag activated gammadelta T cells].

AIM: To explore the role of MAPK signal transduction pathway in killing tumor cells by Mycobacteria tuberculosis antigen (Mtb-Ag) activated human gammadeltaT cells. METHODS: Normal human peripheral blood mononuclear cells (PBMCs) were stimulated with Mtb-Ag and expanded in rIL-2-containing medium to induce gammadeltaT cells. The highly purified gammadeltaT cells were isolated by positive selection with MACS separator. Proportion of gammadeltaT cells was detected by flow cytometry (FCM). The cytotoxicity and CD69 expression of gammadeltaT cells pre-treated with or without PD98059 (Erk inhibitor) or SB203580 (p38 inhibitor) were detected by MTT colorimetry assay and FCM, respectively. RESULTS: The percentages of gammadeltaT cells in freshly isolated PBMCs, Mtb-Ag-activated PBMCs and cells sorted by MACS were 3.56%, 74.63% and 98.20%, respectively. The cytotoxicities of gammadeltaT cells to tumor cell lines (K562 and Raji cells) were inhibited by PD98059. The inhibitory rate of cytotoxicity to K562 cells by gammadeltaT cells (39.27%) was higher than that to Raji cells(26.58%). CD69 expressions on gammadeltaT cells induced by K562 or Raji cells were decreased by pretreatment with 100 mumol/L of PD98059. But SB203580 had no effects on the cytotoxicity and CD69 expression on gammadeltaT cells induced by K562 and Raji cells. CONCLUSION: Erk MAPK pathway, not p38 MAPK, plays an important role in triggering cytotoxicity of gammadeltaT cells. The Erk pathway may affect on the granule exocytosis of gammadeltaT cells more than the Fas/FasL-mediated cytotoxicity of gammadeltaT cells. Moreover, Erk MAPK pathway is also involved in expression of activation molecule CD69 on gammadeltaT cells induced by tumor cells.

Antigens, Bacterial↗

PubMed2005

[In vitro induction and expansion of dendritic cells from rat bone marrow and their characterization].

AIM: To induce and expand dendritic cells (DC) from rat bone marrow in vitro and identify their biological characterization. METHODS: The rat bone marrow cells were collected and cultured for 48 hours and the floating cells were removed. Then IL-4 and GM-CSF were added into the fresh medium. After 2 weeks, the morphological character of the cultured DCs was observed under light microscope and transmission electron microscope. Expressions of MHC class II molecule, B7-1 and B7-2 were detected by flow cytometry. The cultured DCs were co-cultured with allogenic T cells derived from rat spleen. T cell proliferation was measured by MTT colorimetry. RESULTS: The cultured DCs had the typical morphological characterization of DC, and the expression rates of MHC class II molecule, B7-1 and B7-2 were 74.2%, 81% and 76% respectively. The cultured DCs could notably stimulate the proliferation of allogeneic T cells. CONCLUSION: The adherent culture of rat bone marrow cells, and co-culture with IL-4 and GM-CSF can obtain a number of high purity of DCs, which lay the foundation for study on DC's function.

Animals↗

PubMed2005

[Comparison of antitumor effects of dendritic cells modified with different forms of hepatocellular cancer antigens].

AIM: To explore the antitumor effects of dendritic cells (DCs) modified with different forms of hepatocellular cancer antigens. METHODS: DCs were modified with freeze-thawed H22 cell lysate, small molecular H22 peptides and Hsp70-H22 peptide complex, respectively. The cytotoxicity of DCs-activated CTLs to H22 cells was detected by MTT colorimetry. The expression level of IFN-gamma mRNA in splenocytes was detected by RT-PCR. The DCs modified with different antigens were used to immunize mice and then the suppressive effects on the growth of tumor were measured. RESULTS: The DCs modified with H22 peptides alone could not activate CTLs. The ability to activate CTLs by DCs modified with Hsp70-H22 peptide complex was stronger than that by DCs modified with freeze-thawed H22 lysate, with the killing rates of CTLs being 47.3% and 18.3%, respectively. The expression level of IFN-gamma mRNA in 3 groups of splenocytes was consistent with the killing rate of CTLs.The growth of H22 cells in mice immunized with DCs modified with freeze-thawed H22 lysate or Hsp70-H22 peptide complex was suppressed, but the suppressive effect of the latter was stronger than that of the former. The tumorigenesis rate in the Hsp70-22 peptide complex group was only 40%, whereas the tumorigenesis rates in the other two groups were 100%. CONCLUSION: Hsp70-H22 peptide complex is a strong sensitizer for DCs, which participates in immune rejection of tumor through activating CTLs and inducing CD4 (+) T cells to differentiate into Th1 cells.

Animals↗

PubMed2004

[Endothelial nitric oxide synthase gene transfected endothelial cells inhibit smooth muscle cell proliferation and platelet aggregation early in vitro].

OBJECTIVE: To test the function of endothelial cells transfected eNOS gene. METHODS: To test the function of eNOS-overexpressing endothelial cells inhibiting smooth muscle cell proliferation and platelet aggregation in vitro by colorimetry, MTT and (3)H-TdR permeate, respectively, in four different endothelial cells groups. RESULTS: The platelet aggregation is significantly decreasing between the groups of EC and eNOS at the different time (P < 0.05), is 42.2 and 32.6 separately at the time of 120 h. The SMC proliferation is significantly decreasing between the groups of EC and eNOS at the different time (P < 0.05). It is 0.28 and 0.22 by MTT test, 4691 and 3995 by (3)H-TdR permeate separately at the time of 120 h. CONCLUSIONS: eNOS-overexpressing endothelial cells inhibited significantly smooth muscle cells proliferation and platelet aggregation in vitro; which shows powerful effect 48 hours post transfection and lasts up to 120 hours at the least; and were held back by L-NAME.

Animals↗

PubMed2004

[Protection of vascular endothelial cells from TNF-alpha induced injury by quercetin].

OBJECTIVE: To investigate the effects of quercetin (Que) on the TNF-alpha injured human umbilical vein endothelial cell line (ECV-304). METHODS: The experiments were performed in culture of TNF-alpha-induced ECV-304 in vitro. Cell viability was assessed with MTT assay. Lipid peroxidation of ECV-304 was monitored as malonaldehyde (MDA) with a fluorometric assay. NO and SOD of ECV-304 were monitored with colorimetry. The expression of NF-kappaB was detected with immunocytochemistry. RESULTS: TNF-alpha inhibited the ECV-304 prolifteration. Preincubation of ECV-304 with Que for 24h before TNF-alpha exposure increased the cell viability. Que significantly decreased the level of NO and MDA and the expression of NF-kappaB, and increased the activity of SOD in TNF-alpha-induced ECV-304. CONCLUSION: These results demonstrate that Que can produce the protective action on TNF-alpha-induced cultured ECV-304 and its mechanism of action may be related to the decrease of NO, antioxidant effect of lipids. However, antioxidant effect might be exerted through NF-kappaB activation pathway.

Cell Line↗

PubMed2004

[Effect of ligustrazine on injury of vascular endothelial cells ECV-304 induced by hypoxia and lack of glucose].

OBJECTIVE: To investigate the effects of ligustrazine on nitric oxide (NO), malonaldehyde (MDA) production, release of intracellular lactate dehydrogenase (LDH) and membrane fluidity of the injured human umbilical vein vascular endothelial cell line (ECV-304) with hypoxia and lack of glucose. METHOD: The experiments were performed in culture of ECV-304 injured with hypoxia and lack of glucose in vitro. The released LDH of ECV-304 was measured with automatic biochemistry analyse. NO content of ECV-304 was monitored with colorimetry. Lipid peroxidation of ECV-304 was monitored as MDA with a fluorometric assay. The membrane fluidity of ECV-304 was measured with the fluorescence polarization method. RESULT: After culture ECV-304 in hypoxia and lack of glucose for 24 h, the LDH release, MDA production and the membrane fluidity increased significantly and NO level was decreased. Preincubation of ECV-304 with ligustrazine for 24 h reduced LDH release, MDA production, membrane fluidity increasing and increased the level of NO in ECV-304 due to hypoxia and lack of glucose. CONCLUSION: Ligustrazine has protective effect on injury of ECV-304 induced by hypoxia and lack of glucose.

Cell Hypoxia↗

PubMed2004

[Soluble sugar content of clonal plant Neosinocalamus affinis at module and ramet levels].

In this paper, soluble sugar content of Neosinocalamus affinis was measured by anthrone colorimetry at two levels of module and ramet in order to reveal its ecological and physiological adaptability. The results showed that soluble sugar content decreased in the order of leaf > branch > culm. As for soluble sugar content, different modules responded to ramet age and position in different ways. The branch and culm of 1-year-old ramets contented more soluble sugar than those of other four ages, but soluble sugar content in leaf was independent to ramet age. For leaf and culm, lower parts of ramet contented more soluble sugar than middle and upper parts. Under high irradiance, the soluble sugar content of Neosinocalamus affinis leaf was more than that under low irradiance. Seasonal change had significant effect on soluble sugar content. The soluble sugar content of leaf presented a monthly change, with a bottom and upper value in January and in April, respectively, but there was no obvious difference between 2-year-old and 3-year-old ramets. The soluble sugar content of parent ramet leaves was positively correlative to that of daughter ramet in upper and middle parts of ramets. There was no significant difference between ramet ages at ramet level. Hierarchical response of physiological character to environmental changes existed at the module and ramet levels of Neosinocalamus affinis.

Glucose↗

PubMed2005

[Inhibition of CD40 expression on CA46 cell lines by hairpin siRNA eukaryotic expression vectors].

AIM: To construct human CD40 hairpin siRNA eukaryotic expression vectors and investigate their effects on CD40 expression, proliferation and apoptosis of CA46 cells. METHODS: Two DNA oligonucleotides encoding CD40 hairpin siRNAs were synthesized, and cloned into pSilenCircle to construct recombinant plasmid siCD40/pSilenCircle that expressed hairpin siRNAs under the control of pol III U6 promoter. At the same time, antiCD40/pSilenCircle encoding the antisense CD40 RNA and siFly/pSilenCircle encoding siRNA against luciferase were constructed as control vectors. The recombinants were identified by restriction enzyme digestion analysis and sequencing. The recombinant plasmids were transfected into CA46 cells. CD40 expression on CA46 cells and apoptosis were detected by flow cytometry. The proliferation of transfected CA46 cells was detected by MTS colorimetry. RESULTS: (1)siCD40/pSilenCircle, antiCD40/pSilenCircle and siFly/pSilenCircle had been constructed successfully. (2) CD40 expressions on siCD40/pSilenCircle- and antiCD40/pSilenCircle-transfected CA46 cells were significantly reduced, as compared with that on siFly/pSilenCircle-transfected CA46 cells (P<0.01 and P<0.05, respectively). Both hairpin siRNAs and antisense RNA had no influence on the proliferation and apoptosis of CA46 cells. CONCLUSION: Constructed siCD40/pSilenCircle and antiCD40/pSilenCircle can significantly inhibit CD40 expression, but have no significant influence on the proliferation and apoptosis of CA46 cells.

Animals↗

PubMed2005

[The effects of pcDNA3.1-IL-15 transfection on the surface molecules and immune function of murine bone marrow-derived dendritic cells].

AIM: To explore the effects of pcDNA3.1-IL-15 transfected on co-stimulatory molecule expression and immune function on murine bone marrow-derived dendritic cells (DCs). METHODS: Recombinant plasmid pcDNA3.1-IL-15 was constructed and used to transfect DCs. The expression of CD40, CD80 and CD86 on the transfected DCs was analyzed by flow cytometry. Murine splenocytes were stimulated with the transfected DCs. CD4+ and CD8+ T cell subsets in the splenocytes were analyzed by flow cytometry. The proliferation of splenocytes was detected by MTT colorimetry. The IFN-gamma in the culture supernatant of the splenocytes was detected by ELISA. RESULTS: pcDNA3.1-IL-15-transfected DCs expressed higher level of CD40, CD80 and CD86, and induced proliferation of CD4+ T cells and CD8+ T cells in the splenocytes. But the ratio of CD4+/CD8+ T cells was lower than that in the spleen cells stimulated by untransfected DCs or DCs transfected with pcDNA3.1. CONCLUSION: pcDNA3.1-IL-15 can improve the expression of co-stimulatory molecules on DCs and enhance their immune function.

Animals↗

PubMed2005

[Expression of IL-23 gene in murine colon carcinoma cells transduced with recombinant retrovirus and its anti-tumor activity].

AIM: To obtain murine colon carcinoma cell lines stably transducted by recombinant retrovirus encoding mIL-23 gene. METHODS: The retrovirus vector was used to transduce the mIL-23 gene into murine colon carcinoma cells (Colon26) and stable clones expressing mIL-23 (Colon26/IL-23) were obtained by screening with G418. The expression of mIL-23 gene was detected by PCR and RT-PCR. The amount of mIL-23 secreted from Colon26/IL-23 cells and IFN-gamma production by murine splenocytes induced by mIL-23 were determined by ELISA. MTT colorimetry was used to assess the in vitro growth of Colon26/IL-23 cells. BALB/c mice were injected s.c. with Colon26/IL-23 cells to observe their tumorigenicity. RESULTS: Colon26/IL-23 cells were set up. Colon26/IL-23 could secrete mIL-23 which could induce the IFN-gamma production by murine splenocytes. The growth of Colon26/IL-23 cells was similar to that of Colon26 cells in vitro. But the tumorigenicity of Colon26/IL-23 cells was lower than that of Colon26 cells in vivo. CONCLUSION: Colon26/IL-23 cells secreting mIL-23 were obtained and IL-23 had anti-tumor activity.

Animals↗

PubMed2004

[Study on the method for the determination of nitric nitrogen, ammoniacal nitrogen and total nitrogen in plant].

This paper reports the determination of three nitrogen states in plants: nitric nitrogen, ammoniacal nitrogen and total nitrogen. The plant sample was digested by the Kjeldathl method with H2SO4 + H2O2. The solution of the sample was determined by three methods: absorption photometric method, indigotic colorimetry, and UV absorption photometric method after K2S2O8 oxidation. By comparing the results, the relationship among the three N states is that the total N = NH4+ + NO3-. Suitable methods were found for the determination of the elements, thus providing a scientific basis for the selection of suitable methods for chemical element analysis in plants.

Ammonia↗

PubMed2005

[Influence of tangential excision within 24 postburn hours on the local wound inflammatory response in patients with deep partial thickness burn].

OBJECTIVE: To investigate the influence of early tangential excision within 24 postburn hours on local wound inflammatory response and tissue injury in patients with deep partial thickness burn. METHODS: Twelve patients with deep partial thickness burn were enrolled in the study. Tangential excision was performed within 24 postburn hours. The same wound of a patient was divided into three areas for harvesting tissue samples, i.e. pre-operation, post-operation and non-operation areas. For each patient, the biopsies from the pre-operation area were harvested before tangential excision, while those from post-operation and non-operation areas were harvested during 5 to 7 postburn days (PBDs). The contents of IL-8, MPO and MDA in wound tissue were determined by tissue culture and chemical colorimetry, respectively. HE and Mason's staining were employed to assess the degree of necrosis of the wound tissue. RESULTS: The IL-8, MPO and MDA contents in the local wound tissue after tangential excision were 6.83 +/- 1.85 microg/L, 4.07 +/- 0.87 U/g, and 8.94 +/- 5.66 micromol/g, respectively, which were significantly lower than those from non-operation area (P < 0.01). The inflammatory response in the non-operative wound area was distinct with expansion of necrotic tissue area. In contrast, the local inflammatory response of the wound after tangential excision was ameliorated without the enlargement of necrotic tissue area. CONCLUSION: Tangential excision within 24 postburn hours could be beneficial in ameliorating local inflammatory response, and in preventing progressive deepening of the burn wounds, thus it could accelerate wound healing in patients with deep partial thickness burn.

Adult↗

PubMed2004

[Research of anti-proliferation of curcumin on A549 human lung cancer cells and its mechanism].

OBJECTIVE: To investigate the mechanism of anti-tumor effects of curcumin on human lung cancer cell (A549). METHOD: MTT colorimetry method, fluoroscope, FCM combine PI and Annexin V-FITC double pigmentation method and Western blot method were used. RESULT: Under the effect of the curcumin, cell grew against the wall and suspended in the culture liquid, the A549 cell nucleolus were found fragmentated into different size of apoptosis body under fluoroscopy. The cell proliferation were obvious suppressed after treated with different concentration curcumin for 72 hours. The IC50 were 18 micromol/L by using linear regression. The apoptosis induced by curcumin of A549 cell is concentration dependent. With curcumin increased from 5 micromol/L to 30 micromol/L, Annexin-FITC single positive cell (early apoptosis cell) increased from 3.4% to 59.1%. When curcumin concentration reached 40 micromol/L, PI and Annexin V-FITC double positive cell (secondary apoptosis necrosis cell) became major part of cells, and the cell showed G2 phase block. Observed with western blot method, with the increase of curcumin concentration to 10 micromol/L, the expression of PARP increased simultaneously. CONCLUSION: Curcumin can interfere cell growth cycle of A549 cell and suppress cell growth. The suppression effect is concentration dependent. The effect depends not only from the nonspecific cytotoxic but also from induced cell apoptosis.

Adenocarcinoma↗