Objective To investigate the distribution of free Ca 2+ and characterization of Ca 2+ channel in the cultured 11～15 week human embryonic retina. Methods The cells from 11～15 week embryonic retina were dissociated enzymatically and cultured on coated cover slides. Three weeks later, Fluo3, a Ca 2+ indicator, was added in Ca 2+ containing or no Ca 2+ Hepes buffer and incubated with retinal cells for 30 min, meanwhile with or without verapamil, perdipine, or dexamethasone adding. Ca 2+ staining and Ca 2+ transit before and after 10 ?mol/L or 50 ?mol/L KCl stimulation was observed and recorded using Zeiss LM510 confocal laser scanning microscopy. Results In response to K + exposure, a rapid rise of free Ca 2+ in cytoplasma and nuclei of neuron and glial cell was observed, but was repressed with the treatment of verapamil, perdipine, or dexamethasone. The obvious Ca 2+ influx from cytoplasma into nuclei was observed even in the absence of external Ca 2+ . Conclusion L type Ca 2+ channel was expressed and functionally matured in the cultured early stage human embryonic retina.

Objective To explore the clinical causes and preventive measures of complicating ascites of mini-percutaneous nephrolithotripsy (MPCNL).Methods Retrospective analysis of 285 patients with MPCNL for upper urinary tract calculus,which were divided into ascites group and no-ascites group.Results All the procedures were successful.Ascites group of 21 cases,no-ascites group of 264 cases.Univariate analysis showed that the diameter and number of calculus,perfusion pressure,perfusion time,pressure volume of irrigation fluid,preoperative upper urinary tract infection,history of treatment associated with complicating ascites (P＜ 0.05),with age,gender,body mass index no correlation (P＞ 0.05).Logistic regression analysis showed that perfusion pressure,perfusion time,pressure volume of irrigation fluid was independent risk factors after MPCNL concurrent ascites (P ＜ 0.05).Conclusions MPCNL concurrent ascites are closely related to the large perfusion volume,the long operative perfusion time,the high perfusion pressure of irrigation fluid.On the premise of keeping the operative visual field clear,as far as possible to reduce the perfusion pressure,control irrigation fluid-flow rate,reduce the large peffusion volume.These could decrease the coincidence of the ascites.

Objective:To investigate the mechanism of endoplasmic reticulum (ER) stress-induced chemoresistance to cisplatin in gastric cancer cells. Methods:ER stress models were established in both BGC823 and SGC7901 gastric cancer cells. The expression of GRP78, an ER stress marker, was examined by Western blot analysis. Moreover, whether ER stress can decrease the sensitivity of gastric cancer cells to cisplatin and activate P38 was explored by flow cytometry and Western blot analysis, respectively. Whether ER stress-induced chemoresistance to cisplatin can be abrogated by blocking P38 activity in gastric cancer was also elucidated using flow cytometry. Results:GRP78 protein expression markedly increased after treating BGC823 and SGC7901 gastric cancer cells with tunica-mycin (TM) or thapsigargin (TG) for 8, 16, and 24 h (P<0.05), compared with that in the group treated for 0 h. The apoptotic rates of TM-(or TG)-, cisplatin-, and TM (or TG) plus cisplatin-treated groups significantly increased (P<0.05) in both BGC823 and SGC7901 gastric cancer cells compared with the rate in the control group. The apoptotic rate of TM (or TG) plus cisplatin-treated group signifi-cantly decreased (P<0.05) in both BGC823 and SGC7901 gastric cancer cells compared with that of the cisplatin-treated group. Com-pared with the group treated for 0 h, phospho-P38 expression markedly increased after treating BGC823 and SGC7901 gastric cancer cells with TM (or TG) for 8, 16, and 24 h (P<0.05). No difference in P38 protein expression was observed between each group in both BGC823 and SGC7901 gastric cancer cells (P>0.05). Both P38 inhibitors, either SB203580 or PD169316, can inhibit the activation of P38. The inhibition of P38 activity can overcome ER stress-induced chemoresistance to cisplatin in gastric cancer cells (P<0.05). Con-clusion:ER stress can trigger the chemoresistance to cisplatin by activating P38 in gastric cancer cells.

The occurrence of malignant tunors is increasing annually around the world.In every year,8.2 million patients died of malignant tumors in which about 90％ patients died of malignant tumors associated multiple organs metastases.Inhibiting tumor metastasis is the key event to prolong the survival time of patients.With the further research,tumor-vessel associated extravascular tumor metastasis is playing an increasingly important role in the clinical application.This paper summarizes the new developments of tumor-vessel associated extravascular tumor metastasis to provide possible ideas for the therapy of malignant tumors.

Biochemical analysis assays based on colorimetric methods using gold nanoparticles have many advantages including high sensitivity, good selectivity, naked-eyes readout and complex instruments free. These methods have good prospects in applications. The biomolecule assay is highly relative with human health. This review mainly focuses on colorimetric assays applying gold nanoparticles for biomolecules detection.

Objective To investigate the optimal dose of oxycodone for severe pain in cancer patients so as to try to alleviate the suffering rapidly.Methods Totally 135 cases with severe pain in terminal cancer patients were randomly divided into groups A,B,and C,45 cases in each group.Groups A,B,and C were treated with oxycodone hydrochloride sustained-release tablets 20 ～ 140 mg/d,150 ～ 290 mg/d,300 ～ 640 mg/d,respectively.Pain relief and incidence of adverse reactions were evaluated 2 weeks later.Results Pain intensity scores (numerical rating scale,NRS) in 3 groups dropped with statistical difference (P ＜ 0.05).Pain relief rates in groups B and C were statistically significant compared to group A (P ＜ 0.05).Quality of life score (appetite,mental status,sleep,daily activities,and interpersonal life) in group C had statistical significance compared to groups A and B (P ＜ 0.05).Group C had slightly higher incidence of adverse reactions than that of groups A and B without significant difference (P ＞ 0.05).Conclusions Large dose and extra large dose of oxycodone hydrochloride sustained-release tablets can effectively relieve severe pain in advanced cancer patients.Extra large dosage can improve patient's quality of life in spite of the higher incidence of adverse reactions,and patients can still be tolerated well.

Objective To explore the mechanisms of mesenchymal stem cells (MSC) in the treatment of concanavalin A (ConA)-induced acute immune liver injury in mice.Methods MSC were isolated and cultured from bone of the four limbs of three-week-old C57BL/6 mice.The specific surface markers were identified and osteogenic,adipogenic differentiation ability were tested.A total of 15 six to seven-week old C57BL/6 mice were divided into control group,MSC treatment group and phosphate buffer saline (PBS) treatment group,five mice in each group.The mice of MSC treatment group was injected through tail firstly with ConA and then MSC,PBS treatment group was injected through tail firstly with ConA and then PBS,control group was injected through tail with PBS twice.The mice were sacrificed in 14 to 16 hours after injection.The level of alanine aminotransferase (ALT),aspartate transaminase (AST) in peripheral blood were detected and the pathological change in liver tissue was scored by Knodell score system.Activation rate of splenic CD4+ T cells and the proportion changes of T hepler cell (Th)1,Th2,Th17 and regulatory T cells (Treg) were detected by flow cytometry and the ratio of Th17/Treg was calculated.The levels of tumor necrosis factor α (TNF-α),interferon (IFN)-γ and interleukin (IL)-4 in peripheral blood were detected by enzyme linked immunosorbent assay (ELISA).Independent-sample t test was used for comparison between groups of measurement data.Results ALT,AST and Knodell score of MSC treatment group was (174.2± 46.9) U/L,(185.6± 71.6) U/L and 3.4±1.3,respectively,which were better than those of PBS treatment group ((647.0± 118.0) U/L,(749.0± 104.0) U/L and 5.2 ±0.8,respectively),and the differences were statically significant (t =8.33,9.98 and 2.55,all P＜0.05).The activation rate of splenic CD4+ T cell of PBS treatment group was (26.10±2.17) ％,the proportion of Th1 and Th2 in CD4+ T cell was (5.81±0.79) ％ and (5.98± 1.22)％,the ratio of Th17/Treg was 0.29±0.03,the levels of TNF-α,IFN-γ and IL4 in peripheral blood were (1 281.95±88.61) U/L,(1 838.66±196.91) U/L and (1 192.36±163.94) U/L,which were higher than those of control group ((13.74±1.59)％,(1.35±0.17)％,(2.13±0.17)％,0.15± 0.05,(21.71±2.50) U/L,(11.84±1.28) U/L and (24.46±3.96) U/L),and the differences were statistically significant (t=10.26,12.37,7.02,5.30,31.79,15.93 and 20.75,all P＜0.01).There was no statistically significant difference in the activation rate of splenic CD4+T cell between MSC treatment group and PBS treatment group ((26.20±3.09)％ vs (26.10±2.17)％,P＞0.05).However,in MSC treatment group,the proportion of Th1 and Th2 in CD4+ T cell ((1.83±0.52) ％ and (2.75±1.06％)),the ratio of Th17/Treg (0.18±0.02) and the levels of TNF-α,IFN-γ and IL-4 in peripheral blood ((760.71± 73.19) U/L,(742.49±76.46) U/L and (825.76±101.74) U/L) significantly decreased compared with those of PBS treatment group,and the differences were statistically significant (t=9.45,4.48,6.41,10.14,5.56 and 10.22,all P＜0.01).There was no significant difference in the ratio of Th17/Treg between MSC treatment group and control group (P＞0.05).Conclusions The therapeutic effects of MSC on ConA induced acute immune liver injury were through influence splenic CD4+ T cell subsets by decreasing the proportion of Th1 and Th2 and then declining the levels of secreted cytokines such as TNF,IFN-γ and IL-4 in peripheral blood,increasing the proportion of Treg and decreasing the proportion of Th17 and keeping the balance of Th17/Treg.

Objective To evaluate the efficacy and tolerance of monochromatic excimer light (MEL) 308 nm in the treatment of vitiligo. Methods Seventy-seven patients were enrolled in the prospective and open clinical study. Two-hundred and one lesions were subjected to local phototherapy of MEL 308-nm once a week for 3 to 6 months. Results Of these lesions, 86.6% obtained repigmentation at different degrees. The repigmentation was more obvious in lesions on the head, face, neck and trunk than in those on the limbs or extremities. Moreover, patients with generalized and segmental vitiligo got a better improvement than patients with other types of vitiligo. Conclusion MEL 308-nm is effective in the treatment of vitiligo without obvious adverse effects.

The protein product of metastasis-associated in colon cancer 1 (MACC1) gene induces the activation of hepatocyte growth factor/mesenchymal-epithelial transition factor (HGF/c-Met) signaling pathway through transcriptionally activating c-Met gene and upregulating its expression to further promote tumor invasion and metastasis.High level expression of MACC1 is associated with the occurrence and metastasis of a wide variety of human tumors, such as colon cancer, gastric cancer, liver cancer, lung cancer, ovarian cancer etc.In addition, the overexpression of MACC1 is also closely associated with clinical TNM stages and distant metastasis.Thus, MACC1 can serve as an independent indicator of tumor metastasis and prognosis, and become a new target for gene therapy.

Objective To exPlore the effect of mammalian target of raPamycin ( mTOR ) inhibitor eVerolimus on radiosensitiVity of human non_small cell lung cancer cell line in vitro by using eVerolimus to inhibit mTOR signaling Pathway of A549. Methods Human non_small cell lung cancer cell line A549 was subjected to radiation alone or in combination with eVerolimus treatment. The 50%inhibition concentration ( IC50 ) of eVerolimus in A549 cells was detected by methylthiazol tetrazolium ( MTT) assay in vitro. EVerolimus at the 20%inhibition concentration ( IC20 ) was used to Pretreat A549 cells for 24 h. Cells were then irradiated by X_ray with 2,4,6,8 Gy. The cell surViVal fraction was comPuted by clone formation. Cell surViVal curVe was fitted by multitarget one_hit model, and mean lethal dose ( D0 ), dose quasithreshold ( Dq ), surViVal fraction at 2 Gy ( SF2 ), and sensitization enhancement ratio (SER) were calculated. The exPression ofγ_H2AX was determined by Western blotting and then the relatiVe gray Values were analyzed. Results EVerolimus significantly imProVed the sensitiVity of A549 cells to radiation. The D0 , Dq and SF2 of eVerolimus+irradiation grouP were significantly lower than those of irradiation grouP. The SER was 1. 36. The residual amount of γ_H2AX Protein in the eVerolimus + irradiation grouP was significantly higher than that of the irradiation grouP. Conclusion EVerolimus inhibiting mTOR signaling Pathway can increase the radiosensitiVity of A549 cells.