Objective:To assess the prognostic value of methylation of interferon regulatory factor 6 ( IRF6) gene promoter in patients diagnosed with Kidney renal clear cell carcinoma (KIRC). Methods:The primary lesions of fifty KIRC patients who were diagnosed at the First Affiliated Hospital of Nanjing Medical University from January 2016 to January 2020 were collected. The expression of IRF6 protein was determined with an immunohistochemical method. The correlation between the level of IRF6 expression and survival and/or metastasis status was analyzed. The mRNA and protein levels of the IRF6 in KIRC and normal renal tissues were compared by using bioinformatic tools. The difference in the methylation rate of the IRF6 gene promoter between tumor and adjacent tissues was analyzed by searching the online databases. Statistical analysis was carried out for the methylation status of the IRF6 gene promoter region to select those negatively correlated with the overall survival (OS) among the patients. In vitro experiments were conducted with cell lines to verify the correlation between the status of promoter methylation and transcription level of the IRF6 gene. Results:The mRNA and protein levels of the IRF6 gene in KIRC tissues were significantly lower than those of the normal controls, and this was more prominent in patients who had died or developed metastasis. The extent of IRF6 gene promoter methylation in the KIRC tissues was much higher compared with that of the adjacent normal renal tissues. There was a significant negative correlation between the methylation of the IRF6 gene promoter and mRNA level of the IRF6 ( R= -0.52). The higher methylation degree in the IRF6 gene promoter regions cg12034118 and cg16030177, the shorter the OS and worse prognosis in the patients. Only twenty CpG sites in cg12034118 were confirmed to be highly methylated in KIRC cell lines. The transcription level of the IRF6 gene was upregulated in a time- and dose-dependent manner after the treatment with demethylation reagent 5-azadeoxycytidine. Conclusion:The methylation of IRF6 gene promoter in the renal tissues of KIRC patients is closely correlated with the OS. Cg12034118 may provide a promising biomarker for laboratory detection, and its high methylation rate has certain reference value for the prognosis.

Combination of passive targeting with active targeting is a promising approach to improve the therapeutic efficacy of nanotherapy. However, most reported polymeric systems have sizes above 100 nm, which limits effective extravasation into tumors that are poorly vascularized and have dense stroma. This will, in turn, limit the overall effectiveness of the subsequent uptake by tumor cells via active targeting. In this study, we combined the passive targeting via ultra-small-sized gemcitabine (GEM)-based nanoparticles (NPs) with the active targeting provided by folic acid (FA) conjugation for enhanced dual targeted delivery to tumor cells and tumor-associated macrophages (TAMs). We developed an FA-modified prodrug carrier based on GEM (PGEM) to load doxorubicin (DOX), for co-delivery of GEM and DOX to tumors. The co-delivery system showed small particle size of ∼10 nm in diameter. The ligand-free and FA-targeted micelles showed comparable drug loading efficiency and a sustained DOX release profile. The FA-conjugated micelles effectively increased DOX uptake in cultured KB cancer cells that express a high level of folate receptor (FR), but no obvious increase was observed in 4T1.2 breast cancer cells that have a low-level expression of FR. Interestingly, in vivo, systemic delivery of FA-PGEM/DOX led to enhanced accumulation of the NPs in tumor and drastic reduction of tumor growth in a murine 4T1.2 breast cancer model. Mechanistic study showed that 4T1.2 tumor grown in mice expressed a significantly higher level of FOLR2, which was selectively expressed on TAMs. Thus, targeting of TAM may also contribute to the improved in vivo targeted delivery and therapeutic efficacy.

Cerebral Ventricles ; Heart Ventricles ; Humans ; Punctures

Objective    To analyze the clinical characteristics of patients treated with esophagectomy following endoscopic submucosal dissection (ESD) for early stage esophageal cancer or precancerosis and the reasons for esophagectomy. Methods    We retrospectively analyzed the clinical data of 57 patients who were treated with esophagectomy following ESD in West China Hospital and Shanxian Hygeia Hospital from January 2012 through October 2016. There were 42 males and 15 females at age of 65.4 (52–77) years. There were 15 patients of upper thoracic lesions, the middle thoracic lesions in 34 patients, and the lower thoracic lesions in 8 patients. Results    The reasons for esophagectomy included 3 patients with residual tumor, 8 patients with local recurrence, 37 patients with esophageal stricture, and 9 patients with dysphagia, although the diameter was larger than 1.0 cm. The pathology after esophagectomy revealed that tumor was found in 16 patients, including 3 patients with residual tumor and 8 with recurrent tumor confirmed before esophagectomy, and 5 patients with new-found recurrent tumor. Conclusion    In the treatment of early stage esophageal cancer or precancerosis, the major reasons for esophagectomy following ESD include esophageal stricture, abnormal esophageal dynamics, local residual or recurrence.

Objective To investigate the acute effect of local vibration stimulus with low frequency on hand control ability, flexibility, grip strength, and muscle EMG activity. Methods 10 subjects in this study held a vibrator for 20 seconds with isometric contraction at the vibration frequency of 12.5 Hz (vibration group, VIB) or with only isometric contraction (control group, CON). The same procedure measurement, including control ability, flexibility, grip strength, and muscle activity, were recorded both at pre-stimulus (pre-test) and 60-second after stimulus (post-test), respectively. Paired-t test was used to compare the values between pre-test and post-test and the changing rate between groups. Results The control ability and flexibility in pre-post test showed significant improvement after VIB intervention in VIB (P＜0.05). The changing rate of grip strength and EMG in VIB was significantly higher than those in CON (P＜0.05). Conclusions Acute local vibration stimulus with low frequency can be used to improve the hand control ability and flexibility for those who need better hand functional ability in the future.

Objective To investigate the effects of Tai Chi Chuan combined with vibration training on the excitability of α-motorneuron pool and γ-reflex arc. Methods 55 healthy college students were divided into Tai Chi Chuan + vibration training (TAV) group, Tai Chi Chuan training (TAI) group, vibration training (VB) group, and control group (CON) for 8 weeks with 3 times training per week. Each time at pre-and post-training, H-reflex and M-wave were recorded by electrical stimulus induced on soleus muscle. T-reflex was also collected by knocking on the Achill tendon. Results After 8-week training, the ratios of Hmax/Mmax and T-reflex/Mmax in VB group were significantly decreased (P<0.05), while the ratio of T-reflex/Mmax in TAV group was significantly increased (P<0.05), and the change percentage of T-reflex/Mmax was significantly higher than that in VB group (P<0.05). Conclusions Although the vibration training could decrease the resting excitability of α-motorneuron pool, the Tai Chi Chuan combined with vibration training could give the muscle spindle stronger excitement so as to further induce the high excitability of γ-reflex arc. It indicated that the Tai Chi Chuan combined with vibration training is feasible since the neural adaptation around peripheral neuron system could be induced after such kind of training.

OBJECTIVETo investigate the protective effects of selenium on rat hippocampal neurons against ischemia-reperfusion (IR) injury.

METHODSThirty-two rats were randomly divided into sham-operated group, IR group and selenium-treated group, and in the latter two groups, cerebral IR injury was induced by middle cerebral artery occlusion; Na2SeO3 treatment was administer in selenium-treated group. At 14 days after reperfusion, the brain tissues were harvested from the rats and hippocampal neuron injuries were observed by TUNEL and Methylene Blue staining. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and nerve growth factor (NGF) in the hippocampal tissues were measured by ELISA.

RESULTSCompared with IR group, the rats in selenium-treated group showed no significant increase in the expression of m-NGF (P>0.05), but pro-NGF expression was significantly increased (P<0.05) in the hippocampal tissue. Na2SeO3 treatment significantly inhibited the expressions of TNF-α and IL-1β and decreased the apoptosis of hippocampal neurons following cerebral IR injury (P<0.05).

CONCLUSIONSelenium produces antiapoptotic effect to protect the hippocampal neurons following cerebral IR injury possibly not by increasing the level of m-NGF but by decreasing the expressions of the inflammatory factors.

Animals ; Apoptosis ; Brain Ischemia ; metabolism ; pathology ; Hippocampus ; cytology ; metabolism ; pathology ; Interleukin-1beta ; metabolism ; Male ; Nerve Growth Factor ; metabolism ; Neurons ; drug effects ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; pathology ; Sodium Selenite ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism

This study was to investigate the effect of phenylhexyl isothiocyanate (PHI) on drug resistance and sensitivity on K562/A02 cell line to adriamycin (ADM) and to elucidate the possible mechanisms. The inhibition rates of ADM and PHI + ADM on growth of K562/A02 cell line were measured by MTT assay, and K562/A02 cell resistant multiple was calculated. The apoptosis rate of K562/A02 cell line, the changes of intracellular ADM concentrations and MRP1 protein level were detected by flow cytometry (FCM). Intracellular reoxidized GSH level was determined by spectrometric enzyme assay and MRP1 mRNA was assayed by semiquantitative RT-PCR before and after using PHI. The results indicated that the survival rate of K562/A02 cell line decreased with the increasing concentration of PHI. Apoptosis rate increased after treatment in combination with two above drugs, the changes of drug resistance multiple and intracellular ADM level had statistical significance between K562/A02 and K562 cells (p < 0.05), when the concentration of PHI was more than 20 micromol/L. Intracellular GSH level in K562/A02 cell line reduced 5% when 1 microg/ml ADM was used alone, and it increased slightly at first, then decreased when more than 10 micromol/L PHI was used. When more than 20 micromol/L PHI was used in combination with 1 microg/ml ADM, intracellular GSH level in K562/A02 cell line decreased progressively with increasing the concentration of PHI. The expressions of MRP1 mRNA and protein had no statistical significance between K562/A02 and K562 cells (p > 0.05) after or before PHI was used. It is concluded that the cyto-toxicity of PHI to K562/A02 cell line does not associate with the depletion of the intracellular GSH. PHI not only enhances the sensitivity of K562/A02 cell line to ADM, but also partially reverses effect of K562/A02 cell resistance to ADM. ADM combined with PHI can diminish side effect and dosage of ADM.
Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; Isothiocyanates ; pharmacology ; K562 Cells

Objective To identify the different proteins of pancreatic cancer with and without diabetes mellitus (DM) ,so as to provide some clues for understanding the molecular mechanisms of DM in the development .of pancreatic cancer. Methods Samples were divided into pancreatic cancer with or without DM and their normal adjacent tissue with 5 each. The protein expression profiles were analyzed using 2-DE in combination with MALDI-TOF/TOF MS/MS. Western blot was used to verify the expression of a candidate protein CapG. Results Seven proteins were obviously up-regulated in pancreatic cancer with DM(P＜0.05). These proteins were involved in cell motility, metabolism, oxidative stress, etc. Additionally, CapG implicated a functional contribution of cancer invasion and metastasis. Conclusion The results also provide evidence to support the hypothesis that DM plays an important role in the development of pancreatic cancer.

The aim of this study was to investigate the roles of chemokine CCL20 in development of CD4(+)CD25(+) thymocytes by means of fetal thymus organ culture. Fetal mouse thymus lobes were removed at the fetus age of 14.5 days and cultured in complete RPMI 1640 with 20% FBS in vitro. Phenotypes of the thymocytes were analyzed by FACS and the number of cells per lobe was counted. The results revealed that from day 14.5 to day 19, the absolute and relative numbers of the CD4(+)CD25(+) thymocytes varied similarly as their development as in vitro culture at 6 days. Data showed that during the 6 days in vitro culture the CD4(+)CD25(+) cell percentage out of CD4(+) cells was 58.29%, 12.14%, 6.08%, 17.78%, 9.06%, 4.04% and the CD4(+)CD25(+) cell percentage out of CD25(+) cells was 3.75%, 10.81%, 17.20%, 51.93%, 61.64%, 80.06%. All these data indicated similar characters to their development in vivo. Moreover, at interference with CCL20, the percentage of CD4(+)CD25(+) T cells in thymocytes significantly decreased at the 3 and 6 days from 3.24+/-0.18 and 3.96+/-0.24 to 1.27+/-0.11 (p<0.001) and 1.76+/-0.22 (p<0.001) respectively. It is concluded that the development of CD4(+)CD25(+) thymocytes is similar both in vitro and in vivo, interfering with CCL20 significantly downregulate the expression of CD4(+)CD25(+) T cells. The above data may help to understand the development of naturally arising CD4(+)CD25(+) regulatory T cells.
Animals ; Chemokine CCL20 ; pharmacology ; Embryo, Mammalian ; Embryonic Development ; Female ; Male ; Mice ; Mice, Inbred BALB C ; Organ Culture Techniques ; T-Lymphocytes, Regulatory ; cytology ; drug effects ; Thymus Gland ; cytology ; embryology