Objective To investigate the effect of IGF-I on the apoptosis and the expression of survivin through the signaling pathway of PI-3K/Akt in colon cancer cell line SW480.Methods When colon cancer cell line SW480 was culfured,Western blot was used to detect the expression of survivin protein in SW480 cells after they were stimulated by different concentrations of IGF-I for different lengths of time.The phosphorylation of Akt in SW480 cells was determined by Western blot after SW480 cells were stimulated by 100 ng/mL IGF-I with in 2 h.Before and after the specific inhibitor LY294002 was treated in SW480 cells to block the pathway of PI-3K/Akt,the phosphorylation level of Akt protein was detected by western blot,survivin protein by western blot and cell immunofluorescence and the cell apoptosis rates of different treated-groups were measured by flow cytometry.Results IGF-I up-regulated the expression of survivin in a dose-dependent and time-dependent manner in colon cancer line SW480(when treated with IGF-I for 12h,the expression of Survivin reached the peak and when treated with 100ng/ml IGF-I,the Survivin reached the highest expression level);IGF-I actived the signaling pathway of PI-3K/Akt of SW480 cells quickly(the expression of p-Akt proteins in SW480 reached the maximum level when cultured with IGF-I for 15 and 30min,but after that they quickly decreased);IGF-I induced survivin expression through the PI-3K/Akt in colon cancer line SW480;IGF-I could inhibited the apoptosis of SW480 cells through the PI-3K/Akt[use of flow cytometry for detection in the blank,stimulation and block groups showed the SW480 cell apoptosis rate was(5.18?0.415)%,(0.85?0.052)%,(3.15?0.411)% respectirely,with significant differences between the groups(P

Objective To study the effect of high intensity focused ultrasound(HIFU)ablation on microvessel density(MVD)and vascular endothelial growth factor(VEGF)in rabbit hepatcocellular carcinoma (HCC)models.Methods The VX2 t umor cells were planted in the hepatic left-central lobe on 30 rabbits.All animals were divided into two groups randomly:HIFU group(n=25)and control group(n=5).The MVD and the expression of VEGF in the HCC was detected by immunohistochemical SABC methods in control group and 0,1,3.7,16 d after the operation in HIFU group.Results The MVD and the expression of VEGF in the HCC tissue after HIFU treatment in the HIFU group were decreased significantly as compared with those in the control group (P＜0.01).However,there was no significant difference in MVD and the expression of VEGF in the HCC tissue after HIFU treatment in the HIFU group among the different time points(P＞0.05).Conclusions HIFU treatment can effectively destroy the micovessel,repress the neovascularization and reduce the blood supply of the HCC.

Objective To screen the prevalence of celiac disease with serologic markers in the central Chinese population,specifically in patients with chronic diarrhea-predominant irritable bowel syndrome (D-IBS).Methods A total of 282 adult patients with D-IBS were selected based on ROME Ⅲ criteria with 296 age and sex matched consecutive healthy individuals as controls.A gluten-free diet (GFD) was advised in subjects positive for IgA/IgG anti-htTG/DGP antibodies and the serologic antibodies were retested after the GFD.Results Among the 578 study subjects,five D-IBS patients (5/282,1.77％) and two healthy controls (2/296,0.68％) were positive for anti-htTG/DGP antibodies.Among the seven positive cases,one was lost to follow-up and only four were evaluated during GFD therapy for an average of 5.2 months with clinical and/or serological manifestations improved.Conclusions The prevalence of celiac disease may not be uncommon in China.Compared with the healthy population,patients with D-IBS tend to be affected more.Thus,it is significantly important to conduct routine screening for celiac disease in patients with D-IBS.

Pressure overload can result in heart hypertrophy, and induce apoptosis, but the phenomenon of apoptosis during the forepart of cardiomyocyte hypertrophy remains unclear. The aim of this study was to inquire into the process of cardiomyocyte apoptosis during the forepart of cardiomyocyte hypertrophy. We constructed the hypertrophy model by transverse aortic constriction of male SD rat. The apoptosis ratio was detected by TUNEL method and FCM. The result demonstrated that hypertrophy indexes increased with the time of constriction, reached the highest level at day 7, maintained that level, and then decreased at day 91. The apoptosis ratio of cardiomyocyte at day 4 was higher than that at day 0 detected by FCM method, and the ratio at day 7 was markedly lower than that at day 4. After day 28, the apoptosis ratio of cardiomyocyte went up again with the time of constriction after surgery. TUNEL method revealed that positive nuclei were observed in cardiomyocytes exclusively in the left ventricle; the apoptosis ratio increased when constriction continued. TUNEL method confirmed the result of FCM. These data indicate that cardiac hypertrophy is initiated by apoptosis of cardiomyocyte, these two factors(hypertrophy and apoptosis) maintain the balance between growth and death during the early short period of aortic constriction, and when aortic constriction goes on they are involved in the pathogenesis of heart remodeling.

AIM: To investigate the role of mitochondrial ceramidase in mitochondrial functions, especially in the regulation of apoptosis. METHODS: pCDNA3.1/His-MtCDase plasmid, containing mitochondrial ceramidase cDNA sequence, was transfected into K562 cells by liposome, and G418 was used to screen the positive clones. A stable transfected K562 cell line was established and defined as ‘K562TC’. The differences between K562 and K562TC cells in serum withdrawal resistance and Bcl-2 protein expression were evaluated by annexin V/PI test, flow cytometry and Western blotting, respectively. RESULTS: K562TC cells with elevated Bcl-2 protein expression level identified by FCM or Western blotting showed stronger resistance to apoptosis induced by serum withdrawal than their parental cells. Inhibition of mitochondrial ceramidase expression in K562TC cells by its specific antisense oligodeoxynucleotide was correlated with a decrease in Bcl-2 protein level. N, N'-dimethylsphingosine (DMS), a sphingosine kinase inhibitor, depleted intracellular sphingosine-1-phosphate (SPP) production, also abrogated Bcl-2 protein expression in K562TC cells, while exogenous sphingosine-1-phosphate up-regulated Bcl-2 protein level in K562 cells. CONCLUSION: Mitochondrial ceramidase overexpression in K562 cells leads to markedly elevated level of Bcl-2 protein and results in more resistance to serum withdrawal. This effect is initiated not by sphingosine, the direct metabolite of mitochondrial ceramidase, but via sphingosine-1-phosphate, its phosphorylated form, indicating that mitochondrial ceramidase, through its sphingoid metabolite sphingosine-1-phosphate, up-regulates Bcl-2 protein expression in K562 cells.

Objective:To investigate the risk factors and missed diagnosis of intraductal carcinoma of prostate (IDC-P) in patients with metastatic prostate cancer.Methods:The preoperative PSA, prostate MRI, bone scans and lung CT of all patients who underwent prostate biopsy in Department of Urology, Xiangya Hospital, Central South University from January 2018 to July 2020 were reviewed. A total of 261 patients with high suspicion of metastatic prostate cancer were screened for inclusion. Two full-time senior pathologists of urogenital tumors in Xiangya Hospital independently reviewed their pathological sections and detected IDC-P according to the 2016 WHO tumor classification. Diagnostic criteria are defined as malignant epithelial cells filling large acini and prostatic ducts, with preservation of basal cells and solid or dense cribriform pattern/loose cribriform or micropapillary pattern with either marked nuclear atypia or non-focal comedonecrosis.Results:The detection rate of IDC-P was 29.12%(76/261), while the actual reporting rate was only 9.96%(26/261). The results of subgroup analysis including age, PSA level, Gleason score as well as different metastatic sites showed that detection rate of IDC-P was 33.69% in the PSA≥50 ng/ml subgroup, much higher than 17.57% in the PSA <50 ng/ml subgroup ( P=0.0039); And it was 32.33% in the Gleason score ≥ 8 subgroup, much higher than 3.45% in the Gleason score < 8 subgroup ( P<0.01). It was not significantly different in different age subgroups as well as different metastatic site subgroups. These data suggest that PSA ≥ 50 ng/ml as well as Gleason score ≥ 8 may be risk factors of IDC-P.157 samples were stained by immunohistochemistry. The detection rates of IDC-P were 84.21% (16/19) in P63 (+ ) samples, 36.00% (9/25) in ERG (+ ) samples. There were 3 samples with both P63 (+ ) and ERG (+ ), all of which had IDC-P. Conclusions:There is misdiagnosis of IDC-P on prostate needle biopsy in patients with metastatic prostate cancer currently. PSA ≥ 50 ng/ml and Gleason score ≥ 8 are risk factors of IDC-P. Thus, attention should be paid to the possibility of IDC-P in such patients. When the diagnosis is difficult, immunohistochemical staining for ERG and P63 is helpful in IDC-P determination.

OBJECTIVETo observe the effect and side effect of Danzhi Xiaoyao powder (DXP) in treating depression.

METHODSA randomized controlled and double-blinded study was conducted in 63 cases of depression by divided them into the western medicine group (WMG, 31 cases) treated with maprotiline, and the Chinese medicine group (CMG, 32 cases) treated with DXP. The effect of therapy was evaluated before and at the 2nd, 4th and 6th week of the treatment with Hamilton's depressive scale (HAMD), self-rating depression scale (SDS), self-rating anxiety scale (SAS) and the scale for TCM syndrome and symptom differentiation (TCM-SSD), and the side-effect of therapy was assessed with Asberg side-effect scale as well.

RESULTSThere was no significant difference between the two groups in scores of HAMD, SDS, SAS, and TCM-SSD. The markedly effective rate in CMG was 84% and in WMG 87%, showed no significance between them (P > 0.05). The scores of HAMD, SDS and SAS of both groups were remarkably lowered after therapy (P < 0.05). However, the score of Asberg in CMG was lower than that in WMG (P < 0.05).

CONCLUSIONDXP shows the effect equivalent to that of maprotiline, but with obviously less side-effect.

Adult ; Antidepressive Agents ; therapeutic use ; Depressive Disorder ; drug therapy ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Maprotiline ; therapeutic use ; Middle Aged ; Phytotherapy ; Psychiatric Status Rating Scales

Objective To investigate the mechanism of treatment of granulocyte colony-stimulating factor(rhG-CSF),recombinant human interleukin-11(rhIL-11)and recombinant human interleukin-2 (rhIL-2)on hematopoietic injuries induced by 4.5 Gy60 Coγ-ray irradiation in beagles,and to provide experimental evidence for the clinical treatment of extremely severe myeloid acute radiation sickness (ARS).Methods Sixteen beagle dogs were given 4.5 Gy60 Co γ-ray total body irradiation(TBI),then randomly assigned into irradiation control group,supportive care group or cytokines+supportive care (abbreviated as cytokines)group.In addition to supportive care,rhG-CSF,rhlL-11 and rhIL-2 were administered subcutaneously to treat dogs in cytokines group.The percentage of CD34+cells,cell cycle and apoptosis of nucleated cells in peripheral blood were examined by Flow cytometry.Results After 4.5 Gy 60 Co γ-ray irradiation,the CD34+cells in peripheral blood declined obviously(61.3%and 52.1% of baseline for irradiation control and supportive care group separately).The cell proportion of nucleated cells in Go/G1 phase was increased notably(99.27% and 99.49% respectively).The rate of apoptosis(26.93% and 21.29% separately)and necrosis(3.27% and 4.14%,respectively)of nucleated cells were elevated significantly when compared with values before irradiation(P<0.05) 1 d post irradiation.When beagles were treated with cytokines and supportive care,the CD34+cells in peripheral blood were markedly increased(135.6% of baseline).The effect of G0/G1 phase blockage of nucleated cells became more serious(99.71%).The rate of apoptosis(5.66%)and necrosis(1.60%)of nucleated cells were significantly lower than that of irradiation control and supportive care groups 1 d after exposure.Conclusions Cytokines maybe mobilize CD34+cells in bone marrow to peripheral blood,indce cell cycle block at G0/G1 phase and reduce apoptosis,and eventually cure hematopoieticinjuries induced by irradiation.

Objectivc To observe the therapeutic effects of combined cytokines on hematopoietic injuries induced by 4.5 Gy60 Co γ-rays irradiation in beagles,and to provide experimental evidences for the clinical treatment of extremely severe myeloid acute radiation sickness(ARS).Methods 16 beagles were given 4.5 Gy60 Co γ-rays total body irradiation,and then randomly assigned into irradiation control group,supportive care group and cytokines group.In addition to supportive care,recombinant human granulocyte colony-stimulating factor (rhG-CSF),recombinant human interleukin-11(rhIL-11)and recombinant human interleukin-2(rhIL-2)were administered subcutaneouly to dogs in cytokines group.Peripheral blood hemogram was examined once every two days.Bone marrow and peripheral blood were collected to proceed colony cultivation 4 d pre-irradiation and 1 and 45 d post-irradiation.Conventional histopathological sections of sternum were prepared to observe the histomorphology changes. Results After irradiation,the population of all kinds of cells in peripheral blood declined sharply.WBC nadir Was elevated(1.04×109/L,but 0.28×109/L and 0.68×109/L for the irradiation control group and the supportive care group separately),the duration of thrombocytopenia was shortened (24 days,but 33 days for the supportive care groug) and red blood cell counts were maintained in the range of normal values after cytokincs treatment in combination.The colony forming efficiency of haemopoietic stem cells(HSCs)in bone marrow and peripheral blood decreased obviously 1 d post irradiation,but recovered to the level of that before irradiation 45 d post irradiation after supportive care and cytokines treatment.Hematopoietic cells disappeared in bone marrow of animals in irradiation control group,but hematopoietic functions were recovered after cytokines were administrated.Conclusions RhG-CSF.rhIL-11 and rhIL-2 used in combination could elevate WBC nadir,accelerate the recovery of leukocytes,platelets and red blood cells and promote the proliferation,differentiation and maturity of HSPCs left in the body after 4.5 Gy γ-rays total body irradiation,eventually restore the hematopoietic function.Hence,combination of rhG-CSF,rhIL-11 and rhIL-2 could serve as better therapeutic strategy to treat extremely severe myeloid ARS.

Objective To explore the mechanisms of cytokines on acute radiation disease in irradiated beagles.Methods The sera of beagles irradiated with 4.5 Gy γ-rays with cytokines treatment was collected at different time points post irradiation.The two-dimensional gel electrophoresis(2-DE)was used to isolate and compare the differentially expressed proteins in sera.HD-MS was used to analyze the differentially expressed proteins with significance,and the amino acid sequences should be determined. Results High resolution 2-DE gel map was obtained.There were six differentially expressed proteins in sera of irradiated beagles at different time points.Four protein spots were successfully identified by MS.A significant spot was identified as serum amyloid A(SAA)by HD-MS,with relative molecular mass of 13 077 and isoelectfie point of 6.26.Expression of SAA was not found 1 d pre-irradiation and 36 d postirradiation,but increased slightly 1 d(0.2166)and significantly 14 d post-irradiation(0.4577). Conclusions The expression of serum amyloid A was consistent with the process of acute radiation injury,which might indicate the turnover of the disease.