Objective:To construct a signature for identifying active tuberculosis (TB) based on the relative expression orderings (REOs) of gene expression within a single sample.Methods:Using peripheral whole blood samples from 75 active TB and 69 latently infected individuals from four datasets as the training set, and highly stable REO patterns were extracted from the gene expression profile of the two groups of samples. Then, the gene pairs that reversed the REO pattern between the two groups were selected, and each gene pair was ranked in descending order based on their reversal degree. Finally, the top k gene pairs with the highest classification accuracy were selected as the signature for independent dataset validation. Results:A signature composed of seven gene pairs, denoted as 7-GPS, was constructed from the training set. The accuracy rate for 7-GPS to distinguish active TB from latently infected samples was 88.89%, and the accuracy rate for distinguishing active TB from normal samples was 90.09%. In the mixed validation data from different detection platforms, the AUC value for distinguishing active TB from latently infected samples was 0.914 (95% CI: 0.881-0.948), and the AUC value for distinguishing active TB from normal samples was 0.934 (95% CI: 0.904-0.964). In addition, the four genes ETV7, BATF2, ANKRD22 and CARD17P from this signature tended to be highly expressed in peripheral blood samples of active TB, and their expression values were significantly related to the duration of anti-tuberculosis treatment in clinical. Conclusion:The 7-GPS signature is robust and suitable for individualized analysis of a single peripheral blood sample. It has certain clinical application potential.

Objective:To construct a signature for identifying active tuberculosis (TB) based on the relative expression orderings (REOs) of gene expression within a single sample.Methods:Using peripheral whole blood samples from 75 active TB and 69 latently infected individuals from four datasets as the training set, and highly stable REO patterns were extracted from the gene expression profile of the two groups of samples. Then, the gene pairs that reversed the REO pattern between the two groups were selected, and each gene pair was ranked in descending order based on their reversal degree. Finally, the top k gene pairs with the highest classification accuracy were selected as the signature for independent dataset validation. Results:A signature composed of seven gene pairs, denoted as 7-GPS, was constructed from the training set. The accuracy rate for 7-GPS to distinguish active TB from latently infected samples was 88.89%, and the accuracy rate for distinguishing active TB from normal samples was 90.09%. In the mixed validation data from different detection platforms, the AUC value for distinguishing active TB from latently infected samples was 0.914 (95% CI: 0.881-0.948), and the AUC value for distinguishing active TB from normal samples was 0.934 (95% CI: 0.904-0.964). In addition, the four genes ETV7, BATF2, ANKRD22 and CARD17P from this signature tended to be highly expressed in peripheral blood samples of active TB, and their expression values were significantly related to the duration of anti-tuberculosis treatment in clinical. Conclusion:The 7-GPS signature is robust and suitable for individualized analysis of a single peripheral blood sample. It has certain clinical application potential.

Objective:To evaluate the effect of canagliflozin on intrarenal fat content and oxygenation in newly-diagnosed type 2 diabetes patients.Methods:Twenty-three newly-diagnosed type 2 diabetes patients were divided into canagliflozin( n=11) and glimepiride control( n=12) groups .Both groups received MRI scanning with Dixon MRI and BOLD MRI sequence to assess patients′ intrarenal fat content, oxygenation level before treatment and 24 weeks after treatment. Fasting blood glucose, glycosylated hemoglobin, blood uric acid, blood lipids, blood pressure, weight, and other metabolic index were also tested before and after treatment. Furthermore, the relationship between body mass index(BMI) and intrarenal fat content and the correlation between changes in intrarenal fat content and improvement in renal hypoxia were analyzed. Results:No significant differences were found in baseline age, body weight, fasting blood glucose, glycosylated hemoglobin, blood lipid, and serum uric acid between the two groups. There was no significant difference in fasting blood glucose, glycosylated hemoglobin, cholesterol(CHO), low-density lipoprotein-cholesterol(LDL-C), and triglycerides(TG) levels in both groups after 12 and 24 weeks of treatment. The decrease in body weight, blood uric acid level, and diastolic blood pressure from baseline in the canagliflozin group was greater than those in the control group( P<0.05). Two groups of patients with type 2 diabetes at baseline had no obvious difference in intrarenal fat content, and the patients′ BMI showed no obvious correlation with degree of intrarenal fat accumulation. Canagliflozin treatment for 24 weeks could reduce intrarenal fat content, which was higher than that of control group. The R2 * values of renal cortex and medulla in the canagliflozin group decreased from baseline by 19.22% and 22.63% respectively( P<0.05), whereas no significant difference was seen in the glimepiride control group. The decrease of intrarenal fat content in the canagliflozin group was related to the improvement of renal cortex and medulla oxygenation. Conclusion:Canagliflozin can reduce intrarenal fat accumulation and improve renal cortical hypoxia in newly diagnosed type 2 diabetes patients with normal renal function.

A 22-year-old female patient presented with skin flushing in the bilateral legs for 4 years, which gradually spread throughout the whole lower limbs and forearms 6 months ago. Skin examination showed diffuse flushing and dilated capillaries in the lower limbs and both forearms, and the flushing faded after a press. Histopathological examination of the skin lesion on the leg showed hyperkeratosis in a basket-like shape, increased pigmentation in the basal layer, infiltration of the superficial dermis with scattered lymphocytes, with no obvious red blood cell overflow; periodic acid-Schiff staining showed thickened and homogeneous deposits around the blood vessels; immunohistochemical staining showed thickened blood vessel walls and positive staining for type Ⅳ collagen. Diagnosis: cutaneous collagenous vasculopathy.

Objective:To investigate the role of cyclo-oxygenase-2 (COX-2) in breast cancer metastasis and its possible mechanism. Methods: A total of 45 cases of primary breast cancer tissues and brain metastatic breast cancer tissues were collected from patients, who underwent mastectomy in Yunnan Cancer Hospital from October 2015 to April 2018, including 30 cases of primary lesions and 15 cases of brain metastasis. qPCR was used to detect the expression of COX-2 in breast cancer tissues and brain metastatic breast cancer tissues. Recombinant viruses with COX-2 over-expression (LV6-COX2) or COX-2 knockdown (LV3-COX2 shRNA1, LV3-COX2 shRNA2) were transfected into human breast cancer MDA-MB-231 cells; After obtaining the stable expression cell lines, the effect of COX-2 expression on the proliferation of MDA-MB-231 cells was detected by CCK-8, and the effects of COX-2 expression on the migration and invasion of MDA-MB-231 cells were detected by scratch test and Transwell assay, respectively. The mRNAand protein expressions of COX-2 in each group were examined by qPCR and WB, respectively. The effect of COX-2 expression on the expression of EMT-related genes in MDA-MB-231 cells was analyzed by qPCR. Results: The expression of COX-2 in tissues of patients with brain metastases was significantly higher than that in patients with primary breast cancer tissues (P<0.01), and it was correlated with tumor TMN stage in breast cancer patients. MDA-MB-231 cell lines with stable COX-2 over-expression/knockout were successfully constructed. Over-expression of COX-2 promoted the migration and invasion of MDA-MB-231 cells (all P<0.01), and significantly increased the expressions of MMP2, MMP1, N-cadherin and vimentin (all P<0.01), but exerted insignificant effect on cell proliferation. The effect of COX-2 silence exerted the opposite effect and promoted cell proliferation (P<0.05). Conclusion: COX-2 is highly expressed in brain metastatic breast cancer tissues, which may promote the migration and invasion of breast cancer MDA-MB-231 cells by regulating EMT processes.

Objective To study the correlation of cleft lip and palate transmembrane 1 like(CLPTM1L)expression and radiosensitivity of lung cancer cells.Methods Thiazolyl blue tetrazolium bromide(MTT) and cell colony formation assays were used to determine cell growth and survival.Western Blot assay was employed to measure protein expression.Results The results demonstrated a negative correlation between the CLPTM1L expression level and radiosensitivity of lung cancer cells.A lower radiosensitivity in lung cancer cells containing high level of CLPTM1L expression,and vice versa.Enforced expression of CLPTM1L resulted in a significant reduction of radiosensitivity in lung cancer cells irradiated with γ-rays.On the contrary,a marked elevation of radiosensitivity was observed in lung cancer cells transfected with CLPTM1L siRNA.Conclusions CLPTM1L may be a novel target gene in mediating radiosensitivity of lung cancer cells.

AIM: To investigate the protective effect of sodium selenite ( Na2 SeO3 ) on human keratinocytes under ultraviolet-B (UVB) irradiation.METHODS: The cultured HaCaT cells were divided into 4 groups: (1) normal control group;(2) Na2 SeO3 group:pretreated with Na2 SeO3 at doses of 10 nmol/L, 50 nmol/L, 100 nmol/L, 200 nmol/L and 1 μmol/L for 24 h;(3) UVB group: irradiated with UVB at doses of 300, 600 and 900 J/m2; (4) Na2SeO3 +UVB group:after pretreated with Na2SeO3 for 24 h, irradiated with UVB at doses of 300, 600 and 900 J/m2.The cell pro-liferation was detected by MTT assay .The apoptotic rates of HaCaT cells treated with UVB at dose of 300 J/m2 were as-sessed by flow cytometry .RESULTS:Compared with normal control group , the cell proliferation activity in UVB group de-creased significantly ( P<0.05 ) .The cell activity was inversely correlated with the irradiation intensity .No significant difference of the cell activity between Na 2 SeO3 group and normal control group was observed .The cell proliferation in Na2SeO3 +UVB group was higher than that in UVB group significantly (P<0.05).Na2SeO3 at concentration of 100 nmol/L showed the strongest activity to promote cell proliferation .After 300 J/m2 UVB irradiation, the apoptotic rate in Na2SeO3+UVB group decreased significantly ( P<0.05) compared with UVB group .The inhibitory effect of Na 2 SeO3 at concentra-tion of 100 nmol/L on apoptosis was the strongest .CONCLUSION: The damage of human keratinocytes by UVB irradia-tion is in a dose-dependent manner .The photoprotection performance of Na 2 SeO3 reduces the damage of human keratino-cytes induced by UVB irradiation .

Objective To study the nuclear protein association of high-mobility group box-1 (HMGB1) and histone deacetylase 1 (HDAC1),and the effect of interaction on radiosensitivity in human breast cancer cells.Methods The protein-protein interaction was determined by immunoprecipitationWestern blot and glutathione-S-transferase capture assays.Cell growth was examined by MTT (methyl thiazolyl tetrazolium)assay and clonogenic assay.Histone deacetylase activity was analyzed by histone deacetylase assay.Results A significant increase of HMGB1 protein and radiosensitivity was observed in MDA-MB-231 and MDA-MB-468 cells transfected with a pCMV-Tag2B expression vector carrying with a full-length of HMGB1 cDNA.HMGB1 binding to HDAC1 was demonstrated as GST (glutathione Stransferase)-pull down and immunoprecipitation Western blot assay,and the association was elevated by irradiation.An LXCXE motif was required for the HMGB1-HADC1 interaction and HMGB1 radiosensitization.A significant difference of IC50 value was observed,for example,1.8 and 2.2 Gy (wtHMGB1 transfectants,P ＜ 0.05),3.6 and 3.8 Gy (HMGB1/C103F transfectants,P ＞ 0.05),both compared with 3.9 and 4.1 Gy (pCMV-Tag2B transfectants) in MDA-MB-231 and MDA-MB-468 cells,respectively.A specific HDAC1 inhibitor trichostatin A markedly reduced the HMGB1-mediated radiosensitivity,0.5 Gy in the presence of trichostatin A versus 1.8 Gy in absence of trichostatin A in MDA-MB-231 transfectants,1.2 Gy (with trichostatin A) versus 2.2 Gy (without trichostatin A) in MDA-MB-468 transfectants,P ＜ 0.05.Histone deacetylase activity was also detected in immunoprecipitates prepared from these cells with antibodies to HMGB1,and this activity was abolished by the histone trichostatin A.Conclusions These results suggest a previous unanticipated role for HDAC1 in modification of HMGB1-mediated radiosensitivity by its direct interaction with HMGB1.

Objective To evaluate the effect of selenomethionine (Se-Met) against ultraviolet B (UVB)-induced oxidative damage to human HaCaT keratinocytes,and to explore its possible mechanisms.Methods Cultured HaCaT cells were divided into several groups:normal control group receiving no treatment,Se-Met groups treated with Se-Met at concentrations of 1,10,50,100,200 nmol/L and 1 μmol/L for 24 hours respectively,UVB groups irradiated with UVB of 30,60 and 90 mJ/cm2 respectively,Se-Met + UVB groups treated with Se-Met at concentrations of 1,10,50,100,200 nmol/L and 1 μmol/L for 24 hours firstly,then irradiated with UVB of 30,60 and 90 mJ/cm2 respectively.Subsequently,methyl thiazolyl tetrazolium (MTT) assay was performed to estimate cellular proliferative activity,flow cytometry to detect cell apoptosis,colorimetry to evaluate superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and to determine malondialdehyde (MDA) levels.Statistical analysis was carried out by using factorial design analysis of variance (ANOVA),one-way ANOVA and least significant difference (LSD) test.Results Factorial design ANOVA showed that UVB radiation had an inhibitory effect on the proliferative activity of HaCaT cells (F =128.04,P ＜ 0.05),which significantly decreased along with the increase of UVB doses,with significant differences between the three UVB groups (P ＜ 0.05).Se-Met pretreatment also affected cellular proliferative activity (F =5.95,P ＜ 0.05),which was significantly increased in Se-Met (10 nmol/L-1 μmol/L) + UVB groups compared with the UVB groups at corresponding doses (all P ＜ 0.05).There was no significant interaction effect on cellular proliferative activity between UVB radiation and Se-Met pretreatment (F =1.65,P ＞ 0.05).The apoptosis rate of HaCaT cells in the 30-mJ/cm2 UVB group was 31.9％ ± 2.67％,significantly higher than that in the normal control group (4.1％ ± 0.67％,P＜ 0.05) and in the 10-,50-,100-,200-nmol/L and 1-μmol/L Se-Met + 30-mJ/cm2 UVB groups (21.9％ ± 3.72％,17.2％ ± 1.67％,4.6％ ±-0.85％,7.5％ ± 1.86％ and 13.5％ ± 1.95％ respectively,all P ＜ 0.05).Similarly,SOD and GSH-Px activities were significantly weaker (both P ＜ 0.05),while MDA levels were higher (all P ＜ 0.05) in the 30-mJ/cm2 UVB group than in the normal control group;however,there was a significant increase in SOD and GSH-Px activities but a decrease in MDA levels in the Se-Met (10 nmol/L-1 μmol/L) + 30-mJ/cm2 UVB groups compared with the 30-mJ/cm2 UVB group (all P ＜ 0.05).Conclusions Se-Met can reduce UVB-induced oxidative damage to HaCaT cells,likely by enhancing antioxidase activity and decreasing oxygen radicals.

OBJECTIVETo investigate the inhibitory effect of resveratrol on interleukin-1β (IL-1β) production in mesenchymal stem cells (MSCs) exposed to radiation and the action mechanism of resveratrol.

METHODSMSCs were divided into blank control group, radiation group, shRNA interference group, and resveratrol groups. The resveratrol groups were given different doses of resveratrol (50, 100, and 200 µmol/L) before radiation. The secretion and expression of IL-1β was measured by enzyme-linked immunosorbent assay, Western blot, and RT-PCR.

RESULTSCompared with the radiation group, the resveratrol groups had significantly decreased extracellular secretion of IL-1β (t = 83.34, 24.48, and 12.52, P < 0.05 for all) and significantly decreased intracellular expression of IL-1β protein and mRNA (t = 8.695, 14.77, and 13.9, P < 0.05 for all). Compared with those given 200 µmol/L resveratrol alone before radiation, the MSCs treated by SIRT1 silencing and given 200 µmol/L resveratrol before radiation had significantly increased extracellular secretion of IL-1β (t = 18.57, P < 0.05) and significantly increased intracellular expression of IL-1β protein and mRNA (t = 10.24, P < 0.05).

CONCLUSIONResveratrol can significantly inhibit the production of IL-1β in MSCs exposed to radiation, and SIRT1 may play a key regulatory role in the process of inflammation induced by radiation.

Cells, Cultured ; Humans ; Interleukin-1beta ; metabolism ; Mesenchymal Stromal Cells ; drug effects ; metabolism ; radiation effects ; Radiation ; Radiation Dosage ; Stilbenes ; pharmacology