Objective To investigate whether miR-200b suppresses proliferation and induces apoptosis of non-small cell lung cancer cells by targeting DNMT3A. Methods A qRT-PCR was employed for detecting the expression of miR-200b in different non-small cell lung cancer cells and human bronchial epithelial cells. A549 cells were transfected with miR-200b mimics, scramble, DNMT3A-siRNA and control-siRNA, respectively. The scramble and control-siRNA were served the negative control of miR-200b mimics and DNMT3A-siRNA, respectively. Western blot assay was conducted to detect the expression of DNMT3A protein in A549 cells. MTT and Annexin V/propidium iodide staining were employed to detect the proliferation ability and apoptosis rate of A549 cells. The effects of miR-200b mimics and DNMT3A-siRNA on the proliferation and apoptosis rate of A549 cells were compared between groups. Results Results of qRT-PCR showed that the expression of miR-200b was significantly down-regulated in A549, H1299, L78 and H460 cells than that of 16HBE cells. Among them, the most obviously reduction was found in A549 cells (P<0.05). Western blot assay showed that the level of DNMT3A protein was inhibited by restored miR-200b or knock-down DNMT3A in A549 cells. After transfection of miR-200b mimics or knock-down DNMT3A for 48 h, 72 h and 96 h, MTT showed that the OD values, which reflected the optical density of cell proliferation were significantly lower than those in the control group (P<0.05). Annexin V/propidium iodide staining showed that apoptosis rates of A549 cells after transfection of miR-200b mimics or knock-down DNMT3A were (23.33%±0.90%and 20.41%±0.70%), compared with the control group (5.28%± 0.55%and 5.68%±0.47%, P<0.01). Conclusion miR-200b suppresses cell proliferation and induces apoptosis by targeting DNMT3A in non-small cell lung cancer.

Objective To assess the craniotomy in traumatic frontal sinus fracture cerebrospinal fluid( CSF) leaks.Methods Clinical data of 12 traumatic frontal sinus fracture CSF leaks from January 2010 to December 2014, who treated by craniotomy and conservation treatment was invalid were reviewed.Combined typical clinical presenta-tion and basicranial thin-layer computed tomography(CT),made qualitative diagnosis and localization.Craniotomy by bilateral coronary incision and epidural approach was performed.Repairation was mainly for the endocranium and the basicranium.Bone cement was used to reconstruct the osseous defect of the frontal sinus,and then with pedicle periosteal flap coverage.Dural defects was fixed with autogenous fascia.After operation,staying in bed and using anti-biotic for 7-14 days were required,while mannital or lumbar-drainage as needed.Results All 12 cases got posi-tive preoperative CT results.Craniotomy was performed,succeeded without reoperation.None of intracranial infection happened,while 1 case suffered from anosphrasia.Followed up for 3 -12 months, none CSF leaks relapsed. Conclusion Craniotomy by coronary incision,dispose the endocranium and the basicranium for the patients who suf-fered from frontal sinus fracture CSF leaks while conservation treatment is invalid,can obtain satisfied result.

AIM: To study signal transduction pathway of nitric oxide-induced PC12 cell death. METHODS: After PC12 cells were incubated with sodium nitroprusside(SNP), caspase-3 inhibitor Ⅱ plus SNP or p38 inhibitor-SB203580 plus SNP, cell survival rate was quantified by MTT assay and caspase-3 activity was measured with caspase-3 assay kits. RESULTS: SNP induced PC12 cell death in a dose- and time-dependent manner, and increased caspase-3 activity gradually. Both caspase-3 inhibitorⅡand SB203580 reduced cell death significantly, but SB203580 reduced caspase-3 activity significantly. CONCLUSION: NO might induce PC12 cell death through the activation of p38 and caspase-3.

Objective To study the protective effect mechanism of Rosa laevigata Michx (RLM) on cardiotoxicity induced by adriamycin in rats. Method 30 SD rats were randomly into control group, doxorubicin group and RLM groups. The control group was injected with normal saline injection, while the model group was injected with adriamycin intraperitoneally at the dosage of 15 mg/kg every other day. For the RLM groups,1~5 g/kg RLM were given after adriamycin injection. The survival rate, plasma BNP was observed. Apoptosis of cardiomyocyte was detected by instituted-labeled DNA (TUNEL). The activity of GSH-PX, CAT and total SOD in the myocardium tissue were also observed. The expression level of CuZn-SOD , bcl-2 and bax were detected by real-time PCR. Results The survival rate was significantly improved in SD rats treated with RLM compared with that in the adriamycin group (P<0.01). The BNP level was increased when treated by adriamycin (P<0.01), and decreased after RLM administration (P<0.01). RLM could also upregulate the expression of the CuZn-SOD mRNA level, and enhance the activity of GSH-PX, CAT and T-SOD compared with that in adriamycin group. Adriamycin could induce myocardial cells apoptosis, as demonstrated by TUNEL. RLM could inhibit adriamycin-induced apoptosis, bax mRNA expression, and increase bcl-2 expression and bcl-2/bax ratio. Conclusion RLM exhibit some antioxidant activity through many stages, and the anti-apoptosis activity may be related to affect the expression of bax and bcl-2 expression.

Objective To investigate the effect and mechanism of high concentration glucose on cholesterol absorption of human colon cancer epithelial Caco-2 cells.Methods The experimental study was used.(1) CCK-8 detected cell proliferation:the proliferation rate changes of Caco-2 cells were detected by CCK-8 when different concentrations (12.5,100.0,300.0,700.0,1 000.0,1 388.0 mmol/L) of glucose solution effects on Caco-2 cells in order to ensure the half hindering concentration of glucose concentration on Caco-2 cells.(2)Cholesterol absorption of Caco-2 cells was detected:Caco-2 cells were divided into the cholesterol group,cholesterol plus ezetimibe (cholesterol inhibitor) group and blank control group.Cholesterol group:100 μmol/L cholesterol solution and different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution were added.Cholesterol plus ezetimibe group:100 μmol/L ezetimibe,100 μmol/L cholesterol solution and different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution were added.Blank control group:DMEM culture medium and corresponding concentrations of DMSO were added.The cholesterol absorption amounts of Caco-2 cells were measured.(3) The relative expressions of ATP binding cassette G8 (ABCG8),ATP binding cassette G5 (ABCG5),Nickman-Pick CI Like 1 (NPC1L1) and scavenger receptor class B type Ⅰ (SR-B Ⅰ) were examined by Western blot in the different groups.Caco cells were divided into the glucose group,glucose plus ezetimibe group and control group.The different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution were added into the glucose group,different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution and 100 μmol/L ezetimibe were added into the glucose plus ezetimibe group,and 100 μmol/L ezetimibe were added into the control group.The relative expressions of ABCG8,ABCG5,NPC1L1 and SR-B Ⅰ were detected by Western blot.Measurement data were presented as (x) ±s,repeated measure variance analysis was used to perform variation trend test,and t test was utilized to conduct comparisons among groups.Results (1) CCK-8 results showed:proliferation rates of Caco-2 cells with the glucose solution concentration of 12.5,100.0,300.0,700.0,1 000.0 and 1 388.0 mmol/L were 1.380 ±0.043,1.238 ±0.072,0.736 ±0.035,0.336 ±0.021,0.316 ±0.020 and 0.288 ±0.010,respectively,with a statistically significant difference in the proliferation rates (F =11.019,P ＜ 0.05).The half hindering concentration of glucose solution on Caco-2 cells was 283.54 mmol/L.(2)Cholesterol absorption of Caco-2 cells:① the cholesterol absorption amounts of Caco-2 cells with the glucose solution concentration of 5.0,25.0 and 50.0 mmol/L were 0.282 ± 0.042,0.380 ± 0.063,0.390 ± 0.060 in the cholesterol group and 0.042 ± 0.012,0.197 ± 0.015,0.277 ± 0.029 in the cholesterol plus ezetimibe group,respectively,showing a statistically significant difference between the 2 groups (F =55.566,P ＜ 0.05).②There was a statistically significant difference in cholesterol absorption amounts of Caco-2 cells with different glucose solution concentration in the cholesterol group (F =79.117,P ＜ 0.05).The cholesterol absorption amounts of Caco-2 cells with the glucose solution concentrations of 5.0 mmol/L was lower than that with the glucose solution concentrations of 25.0 mmol/L and 50.0 mmol/L,respectively (t =11.207,11.532,P ＜0.05).There was no statistically significant difference in the cholesterol absorption amounts of Caco-2 cells between the glucose solution concentrations of 25.0 mmol/L and 50.0 mmol/L (t =12.389,P ＞ 0.05).③There were statistically significant differences in cholesterol absorption amounts of Caco-2 cells with the glucose concentration of 5.0 mmol/L and 25.0 mmol/L between cholesterol group and cholesterol plus ezetimibe group (t =10.908,10.644,P ＜ 0.05).(3) The results of Western blot showed:① the relative expression of NPC1L1 protein in Caco-2 cells with the glucose solution concentrations of 5.0,25.0 and 50.0 mmol/L were respectively 0.277 ±0.019,0.558 ±0.015,0.576 ±0.003 in the glucose group and 0.057 ±0.002,0.054 ±0.005,0.077 ±0.005 in the glucose plus ezetimibe group,showing a statistically significant difference (F =482.207,P ＜0.05).② The relative expression of NPC1L1 protein of Caco-2 cells with the different concentration of glucose solution in the glucose group were compared,with a statistically significant difference (F =8.112,P ＜ 0.05).There was a statistically significant difference in the relative expression of NPC1L1 protein in Caco-2 cells with the different concentration of glucose solution in the glucose plus ezetimibe group (F =11.708,P ＜ 0.05).③ The relative expression of NPC1L1 protein in Caco-2 cells with the glucose solution concentrations of 5.0,25.0 and 50.0 mmol/L in the glucose group was statistically different from that in the glucose plus ezetimibe group (t =8.112,11.708,13.920,P ＜ 0.05).Conclusion High concentration glucose solution could promote the reabsorption of cholesterol through increasing NPC1L1 protein expression in Caco-2 cells,and increase the risk of suffering from cholelithiasis in diabetes patients.

OBJECTIVE: To optimize reflux extraction technology of total flavonoids from the roots and stems of Angelica sinensis. METHODS: The reflux extraction technology of total flavonoids from the roots and stems of A. sinensis was optimized by Box-Behnken design-response methodology based on single factor test using volume fraction of extraction solvent ethanol, solide-liquid ration, extraction time, extraction times as investigation factors, the content of total flavonoids in extract as evaluation index. RESULTS: The optimal extraction technology of total flavonoids from the roots and stems of A. sinensis was that volume fractions of ethanol were 70％ and 50％; solid-liquid ratios were 1: 40 and 1: 30; extraction time were 1. 3 h and 1. 7 h; The number of extraction times is two times. In verification test, the contents of total flavonoids were 7. 253 6, 25. 144 1 mg/g (RSD= 1. 57％, 1. 49％, n = 3); relative errors of those to predicted value (6. 942 8, 25. 703 5 mg/g) were 4. 28％, 2. 24％. CONCLUSIONS: Optimized extraction technology for total flavonoids from the roots and stems of A. sinensis is simple, reproducible and predictable.

Objective To predict the core targets and action pathways of Hedysari Radix based on UPLC-MS/MS and network pharmacology methods,and to verify the results of network pharmacology by molecular docking and molecular dynamics techniques.This article aims to investigate immune regulation mechanism of effective components absorbed into blood from Hedysari Radix.Methods Qualitative quantification of effective components absorbed into blood from Hedysari Radix were operated by using UPLC-MS/MS technique.The corresponding targets of effective components absorbed into blood from Hedysari Radix were screened by TCMSP and HERB databases.Targets of immune-related disease were obtained through DisGeNET,OMIM,TTD,and MalaCards databases.The network of"components absorbed into blood from Hedysari Radix-immune-related diseases"was then constructed.GO and KEGG enrichment analysis and mapped the PPI network were performed.Molecular docking and molecular dynamics techniques were applied for validation.Results A total of 8 prototype components absorbed into blood,synergistically acting on 101 targets,were identified by UPLC-MS/MS.They mediated 538 biological processes including immune response,positive regulation of gene expression,receptor binding,and cytokine activity.Meanuhile,116 signaling pathways,such as HIF-1,Toll-like receptor,JAK-STAT,T cell receptor,PI3K-Akt,and FoxO etc.were involved.The core targets were MAPK14,PTGS2,MMP9,PPARG,CCND1,etc..The results of molecular docking showed that formononetin and calycosin had strong docking binding activity with MAPK14.And molecular dynamics simulations further demonstrated that the binding between MAPK14 and formononetin or calycosin had good structural stability and binding affinity.Conclusion The results of serum pharmacochemistry,network pharmacology and molecular dynamics were verified to reveal the material basis and mechanism of Hedysari Radix in regulating immunity.The aim of this study is to provide scientific basis for its immunomodulatory mechanism.

[Abstract] Objective: To investigate the expression of galectin-3 in bladder cancer tissues and its correlation with clinicopathological characteristics of bladder cancer patients, as well as to explore its effect on the proliferation, invasion and apoptosis of bladder cancer T24 cells. Methods:Atotal of 104 cases of pathologically confirmed bladder cancer tissues and the corresponding adjacent tissues were collected from the patients treated at the Affiliated Hospital of North Sichuan Medical College from May 2014 to June 2016. Immunohistochemistry staining was used to determine the galectin-3 protein expression in both cancer and adjacent tissues, and the correlations between galectin-3 expression and clinical pathological features were analyzed. siRNA-Gal3 and siRNA-Control were transfected into T24 cell, respectively. The expression of galectin-3 protein was detected by Western blotting, the proliferation of cells was detected by MTT assay; the invasion of cells was detected by Transwell assay; and the cell apoptosis was determined by Flow cytometry. Results: The positive rate of galectin-3 in bladder cancer tissues was significantly higher than that in adjacent tissues (73.1% vs 9.6%, P<0.05). The expression of galectin-3 in bladder cancer was correlated with histological grade, depth of invasion, lymph node metastasis and TNM stage (all P <0.05), but not with sex and age (P>0.05). The expression of galectin-3 was down-regulated significantly by siRNAGal3 (P<0.05). After interference with galectin-3, the proliferation and invasion of T24 cells was significantly decreased (all P<0.05) but the apoptosis was significantly increased (P<0.05). Conclusion: Galectin-3 is over-expressed in bladder cancer and is closely related to the clinicopathological features of bladder cancer patients. Interference of galectin-3 protein expression can inhibit proliferation and invasion and promote cell apoptosis of bladder cancer T24 cells. ··