AIM: To study the expression of nucleostemin (NS) gene in human breast tumor tissues and the relations of NS gene expression level with histological grades, histological types and TNM stages of the tumor.METHODS: Total RNA was isolated from human breast tumor tissue. The methods of electrophoresis and RT-PCR were used in measuring NS gene expression level, and the relations of NS gene expression level with histological grades, histological types and TNM stages of the tumor were analyzed. RESULTS: The results indicated that there was no NS gene expression detected in normal breast tissues, and NS gene expression in malignant breast tumor tissues (P0.05). CONCLUSION: It is suggested that there is no relation of NS gene expression level with histological types of the breast cancer, but there is a marked correlation of NS gene expression level with the histological grades and TNM stages.

Objective　To evaluate the in vitro effect of che motherapeutant on hepatic carcinoma.　Methods　The inhibitory effect of 5-Fu, DDP and 4-PA on Bel-7402 cel ls proliferation was studied by MTT colorimetry.　Resu lts　(1) Bel-7402 was on an active proliferation by the treat ment of 5-Fu and DDP for half an hour, and the proliferation inhibited after 12 hours; but the proliferation was inhibited by 4-PA after half an hour of tr eatment. (2) The inhibitory rates increase with time and were over 50% after 48 ～72 hours of treatment. (3) There was temporarily active proliferation of Bel- 7402 cells in the early period of chemotherapy by 5-Fu and DDP. 　Conclusion　Continuous chemotherapy is effectiv e for the inhibition of Bel-7402 cells proliferation.

AIM: To examine Nucleostemin (NS) expression in tumor cells, and observe the effect of NS specific RNA interference on the cell proliferation in Hela cells. METHODS: Total RNA was extracted from 6 kinds of cultured tumor lines, the NS expression level was measured by RT-PCR and Northern blot. An NS-specific siRNA expression vector was constructed to transfect HeLa cell (NS-siRNA-HeLa), and the proliferation of the cell was observed. RESULTS: NS was highly expressed in 6 kinds of tumor cells. NS expression level in the NS-siRNA-HeLa cells was remarkably reduced, and the percentage of G_0/G_1 cells increased. The neoplasm forming ability in nude mice by the NS-siRNA-HeLa cells was decreased. CONCLUSION: NS is highly expressed among tumor cells. NS-specific siRNA inhibits the entry of the cell cycle into the S phase, and remarkably reduces the proliferation ability of HeLa cells in vitro and in vivo.

Objective To investigate the effect on proliferation and apoptosis by RNA interference to inhibit Nucleostemin(NS) gene expression in gastric carcinoma SGC-7901 cells.Methods The NS-siRNA expression vector was transfected into gastric-carcinoma cells with LipofectamineTM2000 reagent.Then we detected the cell proliferation inhibition ratio by MTT assay,the levels of NS gene expression in all groups by RT-PCR,cell cycle by flow cytometry,cell apoptosis ratio by Annexin-V-FITC/PI kit.Results Compared with that in the control group,cell proliferation in S group decreased;at 24,48 and 72 h the cell proliferation inhibition ratio was 53.21%,71.54% and 87.47%,respectively,the level of NS gene expression reduced in S group.G0/G1 phase cell was 58.34%,S phase cell was 20.68%,and the cell apoptosis was 26.85%.Conclusion RNA interference could substantially inhibit NS gene expression in gastric carcinoma SGC-7901 cells,decrease cell proliferation,arrest cell cycle and increase apoptosis ratio.

Objective To investigate the long-term efficacy of pure transanal total mesorectal excision (PtaTME) for middle-low rectal cancer.Methods The retrospective descriptive study was conducted.The clinicopathological data of 18 patients with middle-low rectal cancer who were admitted to the Sixth Affiliated Hospital of Sun Yat-sen University from July 2014 to August 2016 were collected.There were 7 males and 11 females,aged (58±13) years,with a range from 40 to 84 years.The body mass index was (22±3) kg/m2.All the 18 patients underwent PtaTME.Observation indicators:(1) surgical and postoperative conditions;(2) postoperative pathological examination;(3) follow-up and survival.Follow-up using inpatient reexamination,outpatient examination,and telephone interview were performed to detect anastomotic complications,anal function,urinary retention,sexual dysfunction,survival and tumor recurrence and metastasis once every 3 months within postoperative 6 months,once every 6 months from 6 months to 3 years,and once a year after 3 years up to June 2019.The measurement data with normal distribution were represented as Mean±SD,and the measurement data with skewed distribution were represented as M (range).Count data were expressed as percentages.Survival rates were calculated by the Kaplan-Meier method.Results (1) Surgical and postoperative conditions:18 patients successfully underwent PtaTME,without conversion to open surgery.The operation time,volume of intraoperative blood loss,distance between anastomosis and anal verge,time to first flatus,time to urinary catheter removal,and duration of postoperative hospital stay were (202±68) minutes,50 mL (range,20-400 mL),(4.5± 2.0)cm,2 days (range,2-7 days),3 days (range,2-5 days),and 7 days (range,5-10 days) in the 18 patients,respectively.There was no perioperative complication.Among 18 patients,4 underwent preventive ileostomy.(2) Postoperative pathological examinations:the length of surgical specimens,the number of lymph node dissection,distance from tumor to the distal margin were (11.0±3.0)cm,12±6,and 1.0 cm (range,0.8-3.7 cm),respectively.The 18 patients had complete mesorectal membrane excision,with negative proximal margin,distal margin,and circumferential margin.Tumor pathological staging:there were 2 cases in Tis stage,4 in T1 stage,7 in T2 stage,and 5 in T3 stage;16 in N0 stage,1 in N1 stage,and 1 in N2 stage.Tumor histological classification:2 patients had carcinoma in situ,9 had moderately differentiated adenocarcinoma,and 7 had high-differentiated adenocarcinoma.(3) Follow-up and survival:18 patients were followed up for 34.0-59.0 months,with a median follow-up time of 57.5 months.During the follow-up,4 patients developed grade B anastomotic leakage and were cured after conservative treatment.One patient developed anastomotic recurrence at 2 years after surgery,and no recurrence was found after surgical resection of the recurrent lesion.Four patients with prophylactic ileostomy had the stoma closured,and the anus function was satisfactory after surgery.There was no urinary retention or sexual dysfunction in the 18 patients.Of the 18 patients,17 had tumor free survival after surgery.The 3-year disease-free survival rate was 94.4％,and the 3-year overall survival rate was 100.0％ in 18 patients.Conclusion PtaTME can achieve high quality of specimen,which is safe and feasible for the treatment of rectal cancer.

Objective:To compare peripheral blood tenascin-C (TN-C) level in patients with Kawasaki disease (KD) on admission, after treatment and at recovery, and to assess the potential of TN-C as a novel predictor for coronary artery lesion.Methods:Retrospective study.Blood samples of 44 KD patients [including 21 patients with coronary artery lesions (CAL + group) and 23 patients without coronary artery lesions(CAL - group)], 39 anaphylactoid purpura patients and 36 non-infected and non-vasculitis controls in the Affiliated Hospital of North Sichuan Medical College during January 1, 2018 and November 1, 2018 were collected.TN-C level was measured by enzyme-linked immunosorbent assay.Normally distributed data were compared by the t test; otherwise, they were compared by the Mann- Whitney U test. Pearson product-moment correlation coefficient or Spearman rank correlation coefficient was used to analyze the correlation between TN-C and other laboratory indexes. Results:For KD patients, TN-C levels on admission [(32.0±13.8) μg/L] and after treatment [(33.5±11.4) μg/L] were significantly higher than that at recovery [(23.3±10.8) μg/L](all P<0.01), which was positively correlated with C-reactive protein ( r=0.317, P=0.038), and negatively correlated with sodium level ( r=-0.472, P=0.004). No significant difference in TN-C level was found between CAL + group and CAL - group [on admission: (31.7±15.4) μg/L vs.(32.3±12.5) μg/L; after treatment: (32.2±11.6) μg/L vs.(34.8±11.3) μg/L; at recovery: (22.6±7.3) μg/L vs.(24.0±13.4) μg/L; all P>0.05]. In addition, TN-C level in patients with KD [(32.0±13.8) μg/L] and anaphylactoid purpura [(37.2±18.2) μg/L] was significantly higher than that of control children [(24.0±8.05) μg/L] (all P<0.01). Conclusions:The study findings are able to prove the potential of peripheral blood TN-C as a predictor for coronary artery lesion in KD patients, nor as a maker of vascular injury.Nevertheless, it may be used as an indicator of immune response in the acute phase of KD.

Background: Diabetes can complicate hypertension management by increasing the risk of cardiovascular disease (CVD) and all-cause mortality. Studies targeting diabetes detection in hypertensive individuals demonstrating an increased risk of diabetes are lacking.We aimed to assess the performance of hemoglobin A1c (HbA1c) and its cut-off point in detecting diabetes in the abovementioned population. Methods: Data from 4,096 community-dwellers with hypertension but without known diabetes were obtained from the Study on Evaluation of iNnovated Screening tools and determInation of optimal diagnostic cut-off points for type 2 diaBetes in Chinese muLti-Ethnic (SENSIBLE) study; these data were randomly split into exploration (70% of the sample) and internal validation (the remaining 30%) datasets. The optimal HbA1c cut-off point was derived from the exploration dataset and externally validated using another dataset from 2,431 hypertensive individuals. The oral glucose tolerance test was considered the goldstandard for confirming diabetes. Results: The areas under the ROC curves for HbA1c to detect diabetes were 0.842, 0.832, and 0.829 for the exploration, internal validation, and external validation datasets, respectively. An optimal HbA1c cut-off point of 5.8% (40 mmol/mol) yielded a sensitivity of 76.2% and a specificity of 74.5%. Individuals who were not diagnosed as having diabetes by HbA1c at 5.8% (40 mmol/mol) had a lower 10-year CVD risk score than those diagnosed as having diabetes (P = 0.01). HbA1c ≤ 5.1% (32 mmol/mol) and ≥ 6.4% (46 mmol/mol) could indicate the absence and presence of diabetes, respectively. Conclusions HbA1c could detect diabetes effectively in community-dwellers with hypertension.

OBJECTIVE: To investigate the effect of rapamycin on Parkinson's disease (PD) and its underlying mechanism in mice. METHODS: Sixty SPF adult male C57BL/6 mice were randomly divided into control group, model group and treatment group. 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine(MPTP) was used to induce Parkinson's disease in model group and treatment group. All mice were trained to cross the runway and were subjected to computer-assisted CatWalk. The numbers of tyrosine hydroxylase positive (TH) neurons in the substantia nigra (SN) were assessed by unbiased stereology using the optical fractionator method; protein expression was detected by Western blot analysis; and glutathione peroxidase (GSH-Px), malondialdehyde (MDA) and superoxide dismutase (SOD) were detected by spectrophotometry. RESULTS: In the model group, a decrease in stride rate and an increase in variation of stance and swing were noted by CatWalk system (<0.05 or <0.01); the numbers of TH neurons decreased (<0.01); expression of p-Akt, p-S6K, p-S6 and p-ULK increased (all <0.01); LC3-Ⅱ/Ⅰ ratio decreased (<0.01); MDA level was elevated while the levels of SOD and GSH-PX were reduced (all <0.01). Compared with the model group, after treated with rapamycin, the abnormal behavior including the stride length, variation of stance and swing and step patterns induced by MPTP were all improved (<0.05 or <0.01); the numbers of TH neurons increased (<0.05); the expression of p-Akt, p-S6K, p-S6 and p-ULK was suppressed (all <0.01); the LC3-Ⅱ/Ⅰ ratio was upregulated (<0.05); MDA level decreased while the levels of GSH-Px and SOD increased (all <0.01). CONCLUSIONS Rapamycin inhibits the activation of mTOR pathway, which contributes to protect against the loss of dopaminergic neurons and provide behavioral improvements in mice with Parkinson's disease. These results are partially related to the ability of rapamycin in inducing autophagy and reducing oxidative stress.
Animals ; Behavior, Animal ; drug effects ; Disease Models, Animal ; Male ; Mice ; Mice, Inbred C57BL ; Neuroprotective Agents ; pharmacology ; therapeutic use ; Oxidative Stress ; drug effects ; Parkinson Disease ; drug therapy ; Random Allocation ; Sirolimus ; pharmacology ; therapeutic use ; Substantia Nigra ; drug effects

The aim of this study was to investigate the role of selenoprotein M (SelM) in endoplasmic reticulum stress and apoptosis in nickel-exposed mouse hearts and to explore the detoxifying effects of melatonin. At 21 d after intraperitoneal injection of nickel chloride (NiCl₂) and/or melatonin into male wild-type (WT) and SelM knockout (KO) C57BL/6J mice, NiCl₂ was found to induce changes in the microstructure and ultrastructure of the hearts of both WT and SelM KO mice, which were caused by oxidative stress, endoplasmic reticulum stress, and apoptosis, as evidenced by decreases in malondialdehyde (MDA) content and total antioxidant capacity (T-AOC) activity. Changes in the messenger RNA (mRNA) and protein expression of genes related to endoplasmic reticulum stress (activating transcription factor 4 (ATF4), inositol-requiring protein 1 (IRE1), c-Jun N-terminal kinase (JNK), and C/EBP homologous protein (CHOP)) and apoptosis (B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), Caspase-3, Caspase-9, and Caspase-12) were also observed. Notably, the observed damage was worse in SelM KO mice. Furthermore, melatonin alleviated the heart injury caused by NiCl₂ in WT mice but could not exert a good protective effect in the heart of SelM KO mice. Overall, the findings suggested that the antioxidant capacity of SelM, as well as its modulation of endoplasmic reticulum stress and apoptosis, plays important roles in nickel-induced heart injury.
Animals ; Male ; Mice ; Antioxidants/pharmacology* ; Apoptosis ; Endoplasmic Reticulum Stress ; Melatonin/pharmacology* ; Mice, Inbred C57BL ; Nickel/adverse effects* ; Selenoproteins/genetics* ; Heart/drug effects*

Humans ; Lupus Vasculitis, Central Nervous System/pathology* ; Magnetic Resonance Imaging/methods* ; Diffusion Tensor Imaging/methods* ; Patients ; Lupus Erythematosus, Systemic/pathology* ; Brain/pathology*