Effects of narrow-band UVB combined with tacrolimus ointment in the treatment of vitiligo and its influence on IL-17 and IL-22 in peripheral blood / 中国基层医药

Objective To investigate the effect of narrow-band UVB combined with tacrolimus ointment in the treatment of vitiligo,and its influence on IL-17 and IL-22 in peripheral blood.Methods 100 patients with vitiligo were selected as study subjects,and they were divided into the control group and observation group according to the random number table method,50 cases in each group.The control group was given 0.1% tacrolimus ointment,the observation group was treated with narrow-band UVB combined with 0.1% tacrolimus ointment.After 6 months of treatment,the clinical treatment effect of the two groups was observed.The levels of serum IL-17 and IL-22 were detected by ELISA,the differences between before and after treatment were compared between the two groups.Results The cure rate,effective rate of the observation group were 74.0%,96.0%,respectively,which were significantly higher than 62.0%,84.0% of the control group,the differences were statistically significant(x2=3.125,2.897,all P0.05).After treatment,the levels of IL-17 and IL-22 in the two groups significantly decreased(t=3.454,3.032,all P<0.05),which in the observation group were lower thanthose in the control group(t=3.867,2.665,all P<0.05).Conclusion Narrow-spectrum UVB combined with tacrolimus ointment has good clinical effect on vitiligo,which can effectively improve the local immune response.

Expression difference of FKBP51 in colorectal cancer and normal tissues and its relationship with clinicopathological features / 中国医师杂志

Objective To explore the difference of the expression level of FK506 Binding Protein 51 (FKBP51) in colorectal adenocarcinoma and normal colorectal tissues,and the correlation between FKBP51 expression level and clinicopathological characteristics,and to clarify whether FKBP51 is involved in the occurrence and development of colorectal cancer.Methods By immunohistochemical staining [streptavidin-peroxidase (SP) method] and Western blotting methods tested 31 cases of colorectal cancer tumor tissues and normal colorectal tissues far from tumor 5 cm,and explored the expression level difference of FKBP51.Combined with clinical data of patients,results were analyzed by statistical method x2 test of four case table data.Results The high expression rate of FKBP51 in tumor tissues was 74.19％ (23/31 cases),while the high expression rate of FKBP51 in normal tissue was 9.68％ (3/31 cases).The difference was significant.The expression level of FKBP51 in patients with colorectal carcinoma had no obvious correlation with gender (P =0.771),age (P =0.474),tumor location (P =0.213),degree of differentiation (P =0.318),lymph node metastasis (P =0.124),distant metastasis (P =0.318) and clinical stage (P =0.171);and the tumor size (P =0.049),depth of invasion related (P =0.031),the difference was statistically significant.Conclusions The expression of FKBP51 in colorectal cancer was strong,while weak expression in normal colorectal tissues.With the increase of tumor infiltration and deepening,the expression of FKBP51 became stronger,which indicated that FKBP51 participated in the genesis and development of colorectal cancer,and it might become a new target for individual therapy of colorectal cancer.

Expression and significance of autophagic regulatory factor mTOR and Beclin1 in benign and malignant pleomorphic adenoma / 重庆医学

Objective To explore the expression and significance of main autophagic regulatory factors of mTOR and Beclin 1 in benign and malignant pleomorphic adenoma .Methods The expression of Beclin1 in 35 cases of carcinoma in parotid pleomorphic adenoma(CPA) ,37 cases of benign pleomorphic adenoma(PA) and 20 cases and normal parotid tissues was measured by adopting the immunohistochemical staining method .The expression of p-mTOR and Beclin1 in CPA ,PA and normal parotid tissues was measured by Western blot .Results The positive expression of p-mTOR in normal parotid tissue ,parotid pleomorphic adenoma and parotid pleomorphic adenoma showed the increasing trend ,and the difference among them was statistically significant (P<0 .05) . The Beclin1 level in 3 kinds of tissue showed the descending trend ,the difference among the three tissues was statistically signifi-cant(P<0 .05) .Conclusion mTOR signal factor is over-expressed in CPA tissue ,while the positive expression of Beclin 1 is inhibi-ted in CPA tissue .

Positive expression of phosphorylated mammalian target of rapamycin in gastric caner and its clinicopathological and prognostic significance: a meta analysis / 中华普通外科杂志

Objective To evaluate positive p-mTOR expression and its significance in gastric cancer.Methods Original studies published for the correlation between p-mTOR and clinicopathological parameters as well as prognostic significance were enrolled from Cohrane Library,Pubmed,EMbase database and CBM,CNKI.Analyses were performed by software REVMAN 5.0.Age,gender,TNM stage,lymph node metastasis,type were analyzed using pooled odds ratio (OR) with 95％ confidence interval (CI),and OS were analyzed using pooled hazard ratio (HR) with 95％ CI.Results Seven studies including 2 477 gastric cancer patients were enrolled in the meta analysis.p-mTOR expression was positive in 1 089 of the cases.p-mTOR positive expression was correlated with age,OR =0.74,95％ CI:0.62-0.89,dcpth of invasion OR =0.76,95％ CI:0.60-0.97,lymph node metastasis,OR =1.95,95％ CI:1.59-2.39,TNM stage,OR =0.57,95％ CI:0.38-0.84,type,OR =0.64,95％ CI:0.50-0.83,and OS,HR =1.86,95％ CI:1.60-2.16.Gastric cancer patients with p-mTOR positive expression tend to be younger,had a higher risk of lymphatic invasion,later TNM stage and poor prognosis.Its positive expression had no relation with the gender.Conclusion p-mTOR positive expression is a significant predictor for advanced TNM stage,more lymph node metastasis,intestinal type and poorer OS.

Astragalus polysaccharides inhibit the growth and proliferation of human hepatocellular carcinoma cell via AMPK-mTOR pathway / 中国医师杂志

Objective To investigate the effect of astragalus polysaccharides (APS) on the growth and proliferation of hepatocellular carcinoma cells and its effect on adenosine monophosphate activated pro-tein kinase (AMPK) activity.Methods Hepatocellular carcinoma HepG2 cells cultured for 12,24,and 48 hours were treated with 200,300,and 400 mg/L concentration of astragalus polysaccharides.The cell inhibition rate was detected with methyl thiazolyl tetrazolium (MTT),and apoptosis was observed under the fluorescence microscope.Western blot method was used to measure the expression of total AMPK,phosphorylated AMPK (p-AMPK),and phosphorylate mammalian target of rapamycin (p-mTOR) protein expressions.Results Astragalus polysaccharides of each concentration significantly inhibited the proliferation of human hepatoma HepG2 cells (P ＜ 0.01),and the effect of 300 mg/L concentration astragalus polysaccharides was more significant than that of the 200 mg/L concentration (P ＜0.01);while inhibitory effect of 400 and 300 mg/L Astragalus polysaccharides on the proliferation of human hepatoma cell line HepG2 was not significant difference.We found that Astragalus polysaccharides of each concentration could promote the apoptosis of HepG2 cells,and the effect of 300 mg/L Astragalus polysaccharides was more significant.However,astragalus polysaccharide of 400 mg/L concentration could promote the apoptosis no more than the 300 mg/L concentration,which was observed by fluorescent microscope.Western blot results showed that astragalus polysaccharides could increase the expression of p-AMPK (P ＜ 0.05),and inhibit its downstream protein expressions of p-mTOR (P ＜ 0.05).The proliferation effect of astragalus polysaccharides was weakened after accession of AMPK antagonist compound C on hepatocellular carcinoma cells.Conclusions APS can inhibit the growth and proliferation of hepatocarcinoma cells,and its mechanism is related to the AMPK-mTOR pathway.

Relationship among mTOR levels,regulatory T cells and cytokins in patients with acute coronary syndrome / 天津医药

Objective To assess whether mTOR was activated in peripheral T lymphocytes of acute coronary syndrome (ACS)patients and to investigate the relationship among mTOR,regulatory T cells and cytokins. Methods Patients with acute coronary syndrome (n=32) were selected in ACS group. Meanwhile, patients who complainted of chest pain but were proven to be normal in ECG and in coronary arteriography, were excluded as ACS patients but were diagnosed as Chest Pain Syndrome (CPS) and selected in CPS group (n=28). The expression levels of mTOR, phospho-p70S6KT389 (indicative of mTORC1 activity) and phospho-AKTS473(indicative of mTORC2 activity) were investigated in T cells, which were isolated from peripheral blood of patients in ACS group or CPS group, using Western blot. The proportion of CD4+CD25+Foxp3+Tregs over CD4+cells were evaluated by FACS. And T cell subset related cytokines such as IFN-γ, IL-4, IL-17 and TGF-β1 were exam?ined by ELISA. Then we investigated the relationship among mTOR,regulatory T cells and cytokins by Spearman analysis. Results mTOR and p-p70S6KT389 expression were significantly enhanced in ACS group as compared with those in patients from CPS group (P0.05). By correlation analysis, p-p70S6KT389 expression level positively correlated with IFN-γ, IL-17(rs=0.91,092,P<0.01)and negatively correlated with Tregs and TGF-β1(rs=-0.85,-0.80, re?spectively, P<0.01). Conclusion mTORC1 pathway was activated in peripheral T lymphocytes of ACS patients, and Tregs insufficiency and cytokins imbalance both contribute to the activation of mTORC1 pathway and pathological process of ACS.

Effects of neural stem cells transplantation and FK506 on pathologic and ultrastructural changes of injured spinal cord / 中华创伤杂志

ObjectiveTo investigate the impacts of neural stem cells (NSCs) transplantation on spinal pathology and ultrastructure after spinal cord injury (SCI) in rats and probe into the protective role of tacrolimus (FK506) on neural regeneration.MethodsCompressive SCI at T8 was induced in the adult SD rats,which were randomly assigned to the control group,FK506 group,NSCs group and NSCs + FK506 group.The differences of neural regeneration in each group were compared at days 7,14,28 and 56 after injury by motor evoked potentials ( MEP),HE staining,immunohistochemical staining,ultrastructure observation and image analysis of the myelinated fiber. ResultsThe MEP latency in the NSCs + FK506 group was significantly shorter than that in other groups at day 28 ( P ＜ 0.05 ).HE staining revealed that only local necrosis presented in the NSCs + FK506 group at day 56.More BrdU and NF-200 positive cells were detected with immunohistochemical staining in the other three groups as compared with the control group.Moreover,the positive cells in the NSCs + FK506 group also outnumbered the FK506 group and NSCs group.Electron microscope scan showed edema under the membrane of large myelin sheath in the control group,and classic new myelin sheath and neuraxis in the NSCs + FK506 group at day 56.The regeneration of the nerve fiber in the NSCs + FK506 group was better than that of other three groups (P ＜0.01 ).ConclusionAfter NSCs transplantation for SCI rats,the early combination use of FK506 can improve the pathology and ultrastructure of the regenerative nerve fiber and is conducive to nerve regeneration.

Effect of FK506 on expression of axon guidance cue slit-2 after spinal cord injury in rats / 中华创伤杂志

Objective To investigate the effect of FK506 on the expression of axon guidance cue slit-2 after spinal cord injury(SCI)in rats.Methods A total of 75 adult Sprague-Dawley rats were randomly divided into three groups,ie,sham operation group,SCI group and FK506 treatment group.The SCI model was made by using the modified Allen's technique.Then,the rats were sacrificed and the spinal cord was removed at different time points(at days 1,3,7,14 and 28)for detection of the expression of slit-2 by means of reverse transcription polymerase chain reaction(RT-PCR)and immunohistochemistry.Results The expression of slit-2 changed with time.The expression of slit-2 could be detected at day 1 after SCI,reached the highest at day 7 and then decreased gradually,with higher expression level in the injury group compared with treatment group(P ＜ 0.05).Conclusion Following spinal cord injury,administration of FK506 can up-regulate the expression of slit-2 and may exert important effect on the guidance of the axon regeneration.

Action of tacrolimus on Wistar rat kidneys implanted with Walker 256 carcinosarcoma / Estudo da ação do tacrolimus em rins de ratos Wistar implantados com carcinossarcoma de Walker

PURPOSE: To evaluate the development of Walker 256 tumor in male Wistar rats treated with tacrolimus using an experimental kidney tumor model. METHODS: 40 male Wistar rats were divided into four groups: Tumor group (TU) (n=10), Tacrolimus-Tumor group (TT) (n=10), Tacrolimus group (TC) (n=10) and Control group (C) (n=10). Treatment with tacrolimus was performed in groups TT and TC. Under anesthesia, the right kidney of each animal of TU and TT was accessed through a supraumbilical incision and inoculated with a 0.1mL solution containing 2x10(6) tumor cells (Walker 256 carcinosarcoma tumor cells). Group TC was treated with a saline solution. All the animals of groups TC and TT were treated with tacrolimus (5mg/kg/day) by gavage for 15 days. TU group animals received saline by gavage for 15 days. On the 15th postoperative day, all animals were submitted to euthanasia and blood sampling for analysis of serum creatinine (Cr) and blood urea nitrogen (BUN). Abdominal gross examination was performed, the right kidney removed and prepared for histological analysis by hematoxylin-eosin staining. The resulting data were submitted to statistical analysis by ANOVA. RESULTS: Statistical significance was found when comparing creatinine level between groups TU, TT and TC -TT group culminated with a marked increased in creatinine levels (Cr=1.013 ± 0.3028 mg/mL), TU group (Cr=0.5670 ± 0.03536 mg/dL) P=0.00256, TC group (Cr =0.711 ± 0.1653 mg/mL) P= 0.02832. Statistical significance was found when comparing BUN levels in TT group (71.32 ± 17.14 mg/mL), compared with TU group (45.83 ± 5.046 mg/dL), P=0.000318. There were no statistically significant differences between groups TT and TC (61.23 ± 9.503 mg/mL) P=0.7242. Histological analysis showed a poor evolution in TT group with multiple foci of hemorrhage and cortical invasion by the Walker tumor. CONCLUSION: The Tacrolimus-treated group developed a more aggressive tumor and a drug-related nephrotoxic effect.

OBJETIVO: Avaliar as alterações na evolução do carcinosarcoma 256 de Walker, inoculado no rim de ratos Wistar, sob tratamento imunossupressor com o tacrolimus. MÉTODOS: Foram utilizados 40 ratos Wistar, machos divididos em quatro grupos de 10: grupo Tumor (TU), Tumor-Tacrolimus (TT), Tacrolimus (TC) e Controle (C). Os ratos dos grupos TU e TT foram inoculados com 0,1 mL de solução contendo 2x10(6) células do tumor de Walker no parênquima do rim direito. Os dos grupos TC e TT receberam tratamento com tacrolimus na dose de 5mg/kg de peso, via gavagem orogástrica durante 15 dias. Os ratos do grupo TU receberam solução salina isotônica pH 7,2. Ao 15º dia de evolução, todos foram submetidos à eutanásia. Amostras de sangue eram coletadas para dosagem de creatinina (Cr) e uréia (Ur) e posteriormente realizada nefrectomia para avaliação histológica. RESULTADOS: As dosagens séricas de creatinina foram maiores no grupo TT (Cr = 1,013±0,3028 mg/mL), que diferiu significantemente dos grupos TU (Cr=0,5670 ± 0,03536 mg/dL) com p=0,00256 e do TC (Cr=0,711 ± 0,1653 mg/mL) com p=0,02832. As dosagens séricas de uréia foram maiores no grupo TT (71,32 ± 17,14 mg/mL), que diferiu significantemente do grupo TU (45,83 ± 5,046mg/dL) com p=0,000318, mas comparado ao grupo TC (61,23 ± 9,503 mg/mL) não houve diferença significante (p=0,7242). No inventário da cavidade abdominal dos grupos TU e TT, observou-se presença macroscópica de tumor em todos os rins direitos; não foram evidenciadas efusões ascíticas, formação de bridas ou metástases tumorais em outros órgãos ou tecidos adjacentes aos rins direitos. CONCLUSÃO: O tacrolimus exerceu efeito nefrotóxico e induziu exacerbação do crescimento do tumor de Walker 256, quando implantado no rim de ratos Wistar.

Expression and significance of phosphorylation-mammalian target of rapamycin in endometrial carcinoma / 中国医师进修杂志

Objective To investigate the expression of phosphorylation-mammalian target of rapamycin (p-mTOR) in endometrial carcinoma. Methods The expressions of p-mTOR protein in tissues from 45 cases of endometrial adenocarcinoma, 7 endometrial atypical hyperplasia, and 6 normal endometrium were detected by immunohistochemistry SP method. Results The expression of p-mTOR protein maily restricted to cytoplasm. Compared with normal endometrium, the expression of p-mTOR protein in the cases of endometrial atypical hyperplasia and endometrial adenocarcinoma was significantly up-regulation(2.36 ± 0.76vs 6.21 ± 1.19, 15.82 ± 2.64)( F = 11.37, P ＜ 0.05 ). There was significant difference of p-mTOR protein in different histology class in endometrial adenocarcinoma (F = 8.27, P ＜ 0.05), but there was no significant difference of p-mTOR protein in different pathological grade in endometrial adenocarcinoma (P ＞0.05).Conclusion The expression of p-mTOR protein may participate in the occurrence and development of endometrial carcinoma.

Study on the changes of mammalian target of rapamycin with aging in rat kidneys / 中华老年医学杂志

ObjectiveTo investigate the changes in mammalian target of rapamyein (mTOR) with aging in rat kidneys.MethodsMale Wistar rats at the ages of 3, 12, 24 months were used for this study. Therenaltissuesandmesangialcellswereprocessedfor senescenceassociated β-galactosidase (SA-β-gal) staining. The expression and location of roTOR in kidneys and mesangial cells were studied by immunohistochemistry and immunocytochemistry, respectively. The mRNA and protein levels of the roTOR and p-roTOR were detected by Western blot assay and RT-PCR,respectively.ResultsThe expression of neutral β-galactosidase activity was increased in kidneys and mesangial cells with advancing age. Percentages of SA-β-gal staining positive ceils were (11.9±3.6)% versus ( 39.0±4.0)% versus ( 86.9±7.4) % in young, middle and aging glomerular mesangial cells (P<0.05). The mTOR staining appeared in the mesangial matrix and interstitium in kidneys, while the mTOR protein showed localization in cytoplasm and nucleus in mesangial cells. The staining intensity of mTOR in kidneys and mesangial cells in aged rats was markedly increased as compared to that in young and middle aged rats (P<0.05). The mRNA level of roTOR was significantly increased in kidneys and mesangial cells of agedrats versus young and middle aged rats,meanwhile, the roTOR and p-mTOR protein expressions were dramatically increased with advancing age (P<0.05 ).ConclusionsmTOR expression is increased with aging, which may play an important role in the aging process of kidneys.

Study on protection of Bcl-2 on islet cells cultured in immunosuppressive drugs / 中国医师杂志

Objective To investigate the toxic effect of immunosuppressive drugs on apoptosis of rat pancreatic islet cells in vitro and the protective action of Bcl-2.Methods Islet cells expressing Bcl-2 and the control islet cells were cultured at different concentrations of tacrolimus and the apoptosis rate of islet cells and insulin accumulation in the culture medium were detected after 48h.Results Low and high concentrations of tacrolimus induced the apoptosis of islet cells and decreased insulin secretion.The Bcl-2 inhibited the apoptosis of islet cells induced by tacrolimus and improved the insulin secretion.Conclusion Tacrolimus may directly damage to isolated rat islet cells and the expression of Bcl-2 can protect the cells from the damage of immunosuppressive drugs.