Background:Fenretinide,which is capable of generating reactive oxygen species( ROS ),has emerged as a promising antineoplastic agent based on numerous in vitro and in vivo studies and clinical chemoprevention trials. Preliminary studies showed that fenretinide could induce apoptosis in human hepatocellular carcinoma( HCC)cells in vitro, however,the precise mechanism was not clarified. Aims:To elucidate the effect of ROS on apoptosis of human HCC cells induced by fenretinide and the underlying mechanism. Methods:Human HCC cell line Huh-7 was treated with antioxidant vitamin E,fenretinide or their combination,respectively. ROS in live cells was evaluated by confocal microscopy and flow cytometry;cell viability and apoptosis were assessed by CellTiter-Glo Luminescent Cell Viability Assay Kit and Caspase-Glo3/7 Assay Kit;expression and intracellular localization of nuclear receptor Nur77,as well as expression of stress-induced transcription factor GADD153 were measured by immunofluorescence staining and Western blotting,respectively. Results:Vitamin E pretreatment fully blocked the fenretinide-induced ROS production. In Huh-7 cells pretreated with vitamin E,cell apoptosis induced by fenretinide was significantly reduced(P<0. 05). Furthermore,effect of vitamin E pretreatment was noteworthy on reducing fenretinide-induced GADD153 expression, while no significant impact on fenretinide-induced Nur77 expression and translocation was observed. Conclusions:Elimination of ROS by vitamin E can abrogate the pro-apoptotic effect of fenretinide on Huh-7 cells,which indicates the participation of ROS in fenretinide-induced apoptosis of human HCC cells. Its mechanism might be associated with induction of GADD153 protein expression.

PurposeTo evaluate the significance of ultrasonic elastography strain ratio, MRI and the combination of both in diagnosis of breast tumor.Materials and MethodsFifty-four cases with single breast tumor underwent preoperative ultrasound elasticity imaging and MRI. Accuracy of ultrasound elastography strain rate ratio (SRR) of the tumor and surrounding normal breast tissue was measured by quantitative ultrasound elastography, and its combination with MRI were analyzed. ResultsThere was signiifcant differences on SRR between the benign group and the malignant group (2.24±1.28vs 4.96±1.73, t=2.648,P<0.05). Optimal threshold of ultrasonic elastography SRR in differential diagnosis of breast benign from malignant tumor was 2.41 determined by ROC curve. The accuracy of SRR, MRI and the combination of both in differentiating benign from malignant breast tumor was 81.48% (44/54), 85.19% (46/54) and 96.30%(52/54), respectively. There was no statistic difference between SRR and MRI in diagnostic accuracy (χ2=0.267,P>0.05). Combined both had higher diagnosis accuracy when compared with SR and MRI separately (χ2=6.000 and 3.967,P<0.05).Conclusion Ultrasonic elastography strain ratio is accurate and objective in differentiating benign from malignant breast tumors. It is a valuable quantitative index in clinical practice. Moreover, SRR combined with MRI can reduce the misdiagnosis rate.

OBJECTIVETo analyse retrospectively 3-tunnels core decompression with implantation of bone marrow stromal cells(bMSCs) and decalcified bone matrix (DBM) for treatment of femoral head necrosis in early stages, and to study its therapeutic results and indications.

METHODSAccording to the University of Pennsyvania system of classification and staging,to treat the patients with 3-tunnels core decompression with implantation of bone marrow stromal cells (bMSCs) and decalcified bone matrix (DBM) for treatment of early femoral head necrosis. Among the total 87 patients (103 hips), the male was 71 (86 hips) and the female 16 (17 hips) with the average age of 47 years old. The average course of disease was 1.3 year. The effect was evaluated by the clinical symptoms, the Harris' scores and the manifestations of radiology before and after operation.

RESULTSAll the patients were followed up more than 2 years (average 26 months). The average Harris' scores increased from preoperation 47.3 to postoperation 74.0. The average rate of excellent and good results was 75.7%. The rate of excellent and good was 88% (22/25) in type I, 78.7% (37/47) in type II and 61.3% (19/31) in type III. No severe complications were observed.

CONCLUSIONThree-tunnels core decompression with implantation of bMSCs and DBM not only removal the focus of disease, but also use DBM the induction of bone and autologous hone marrow stem cells to differentiate the functions of the femoral head can be resumed loading structure, eliminate pain,improve joint function. It is an effective method for early femoral head necrosis.

Adult ; Aged ; Bone Marrow Transplantation ; Bone Matrix ; transplantation ; Decompression, Surgical ; Female ; Femur Head Necrosis ; surgery ; Follow-Up Studies ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Middle Aged ; Retrospective Studies ; Transplantation, Autologous

OBJECTIVETo investigate the in vivo inhibition of extract of Fructus lycii (FL) on the expressions of cathepsin B (Cat B) and cystatin C (Cys C) in high-fat diet and hydroquinone (HQ) induced model mice with age-related macular degeneration (AMD), and to explore the in vitro effects of lutein and zeaxanthin on hydrogen peroxide (H2O2,) induced expressions of matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinase 2 (TIMP-2) on ARPE-19 cells.

METHODSFifty female 8-month-old C57BL/6 mice were recruited in this research. Ten mice fed with regular diet was taken as the age control group. The rest 40 mice were fed with high fat diet for 6 months, followed by adding HQ (0. 8%) in the drinking water for 3 consecutive months. Then the modeled mice were randomly divided into the model control group (n =10), the high (at the daily dose of 3.75 g/kg), middle (at the daily dose of 2.50 g/kg), and low dose (at the daily dose of 1.25 g/kg) FL groups, 10 in each group. The extract of FL at each dose was respectively administered to mice by gastrogavage for 3 successive months. By the end of the experiment, the mice were killed and their eyeballs were removed. The protein expressions of Cat B and Cys C were observed by immunohistochemical assay. The mRNA and protein expressions of Cat B and Cys C were detected by real-time PCR and Western blot respectively. The drug concentrations of H2O2, lutein, and zeaxanthin were screened and detected using the activity of cell proliferation. The protein expressions of MMP-2 and TIMP-2 were detected using Western blot.

RESULTSCompared with the age control group, the mRNA and protein expressions of Cat B and Cys C were significantly higher in the in vivo model control group (P <0.05, P <0.01). The mRNA expressions of Cat B and Cys C were weaker in the middle and high dose FL groups than in the model control group (P <0. 05, P <0. 01). In in vitro cells, lutein and zeaxanthin could down-regulate the protein expressions of MMP-2 and TIMP-2 in H202 induced ARPE-19 cells (P <0. 05, P <0. 01).

CONCLUSIONSExtract of FL could down-regulate the high protein expressions of Cat B and Cys C in high-fat diet and HQ induced model mice. Lutein and zeaxanthin could down-regulate the protein expressions of MMP-2 and TIMP-2 in H202 induced ARPE-19 cells.

Animals ; Cathepsin B ; metabolism ; Cystatin C ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hydrogen Peroxide ; Lutein ; pharmacology ; Macular Degeneration ; prevention & control ; Matrix Metalloproteinase 2 ; metabolism ; Mice ; Mice, Inbred C57BL ; Pigment Epithelium of Eye ; drug effects ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism ; Xanthophylls ; pharmacology ; Zeaxanthins

Objective To study the quality of life and its influential factors among HIV positive population. Methods Quality of life was evaluated among 2608 HIV positive population by WHOQOL (Chinese Version) to calculate the total and four domains' scores of quality of life. Multiple linear regression model was used to analyze the relationship between the factors and the scores of the four domains and the total score of quality of life. Results Physical,psychological,social,environmental and the total scores of the HIV positive population were 12.96±1.94,11.79± 1.19,13.79±2.44,12.40±1.93 and 51.02±6.03,respectively. Females had a higher scores than males in the four domains and the total score of quality of life. Through a multiple linear regression model,we found that the influential factors would include gender,age,occupation,family conflict and appetite etc. Conclusion People living with HIV had higher scores in social domain,but lower scores in psychological domain,suggesting that psychological intervention should he strengthened.

OBJECTIVETo retrospectively analyze the clinical value of diffusion-weighted MR imaging in the detection of prostate cancer in suspected patients.

METHODSBetween January 2009 and December 2010, the 551 patients suspected as prostate cancer underwent prostate biopsy. Patients in group A were accepted to a transrectal ultrasound (TRUS) guided transrectal prostate biopsy (n = 410), while patients in group B were accepted to a diffusion weighted imaging (DWI) and TRUS jointly guided transrectal prostate biopsy (n = 141). The two groups were divided into 4 subgroups by prostate specific antigen (PSA) < 10 µg/L, 10 µg/L ≤ PSA < 20 µg/L, 20 µg/L ≤ PSA < 50 µg/L and PSA ≥ 50 µg/L. Then, the diagnostic rates of prostate biopsy guided by combination of DWI and TRUS with only TRUS were compared.

RESULTSThe diagnostic rate of patients with PSA < 10 µg/L, 10 µg/L ≤ PSA < 20 µg/L, 20 µg/L ≤ PSA < 50 µg/L and PSA ≥ 50 µg/L were 12.1%, 31.1%, 48.0%, 91.2% in group A, and 23.7%, 35.5%, 66.7%, 96.3% in group B, respectively. In the patients with PSA less than 10 µg/L, there were significant differences in diagnostic rate between the two biopsy techniques (χ(2) = 4.405, P < 0.05).

CONCLUSIONThe combination of DWI and TRUS showed the potential to guide biopsy to cancer foci in patients suspected as prostate cancer. For patients with PSA < 10 µg/L, a DWI and TRUS jointly guided transrectal prostate biopsy was recommended.

Biopsy, Needle ; methods ; Endosonography ; Humans ; Magnetic Resonance Imaging ; Male ; Prostate ; diagnostic imaging ; pathology ; Prostatic Neoplasms ; diagnosis ; pathology ; Retrospective Studies

OBJECTIVETo study the mechanism underlying the effect of combined use of cyclonpamine and hydroxycamptothecin in inducing the apoptosis of human oral squamous cell carcinoma cell line (OSCC) HSQ-89.

METHODSCCK8 assay was used to investigate the inhibitory effect of cyclopamine on HSQ-89 cells. Flow cytometry (FCM) was employed to examine the cell apoptosis following combined treatment with cyclonpamine and hydroxycamptothecin. Reverse transcription polymerase chain reaction (RT-PCR) was applied to detect the mRNA expressions of Bcl-2, Bcl-xl, and Bid in HSQ-89 cells after the treatments.

RESULTSCombined treatment with cyclonpamine and hydroxycamptothecin significantly inhibited the cell proliferation compared with hydroxycamptothecin treatment alone, also resulting in a significantly higher apoptosis rate of the cells (P<0.05). The mRNA level of Bcl-2 was significantly decreased after the treatments, especially after the combined treatment. Cyclopamine produced no significant effect on the mRNA levels of Bcl-xl and Bid in the cells.

CONCLUSIONThe combined use of cyclopamine and hydroxycamptothecin significantly down-regulates the expression on of bcl-2 to induce the apoptosis of human OSCC cell line HSQ-89.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Camptothecin ; analogs & derivatives ; pharmacology ; Carcinoma, Squamous Cell ; pathology ; Cell Line, Tumor ; Drug Synergism ; Humans ; Mouth Neoplasms ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Veratrum Alkaloids ; pharmacology

OBJECTIVETo explore the effects of epigallocatechin-3-gallate (EGCG) on the proliferation of human oral epithelial cancer cell line KB cells and the molecular mechanisms.

METHODKB cells were treated with various concentrations of EGCG for 24 or 48 h. MTT assay was used to test the cell viability. The changes of cell cycle in KB cells treated with EGCG for 48 h were analyzed using flow cytometry. The expressions of cyclin A, cyclin D1 and cyclin E were detected by RT-PCR and Western blotting.

RESULTThe viability of KB cells treated with various concentrations of EGCG (25, 50, 100, 200, 400, and 800 micromol/L) for 48 h were decreased to (85.4-/+2.4)%, (80.4-/+2.8)%, (51.5-/+4.5)%, (30.2-/+1.9)%, (25.3-/+1.5)%, (20.0-/+1.1)%, respectively, showing significant difference from that of the control group [(100.0-/+2.2)%, P<0.05). EGCG decreased the viabilities of KB cells in a dose-dependent manner. Flow cytometry demonstrated that treatment with EGCG significantly increased the cell percentage in sub-G1 phase, which was (73.5-/+4.4)% after a 48-h EGCG treatment, significantly different from that in the control group [(47.3-/+3.5)%, P<0.05). EGCG-induced G1 phase arrest was correlated to the down-regulation of cyclin A and cyclin E.

CONCLUSIONEGCG inhibits the proliferation of KB cells by inducing G1 phase arrest, which involves the downregulation of cyclin E.

Catechin ; analogs & derivatives ; pharmacology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cyclin E ; metabolism ; Flow Cytometry ; G1 Phase Cell Cycle Checkpoints ; drug effects ; Humans ; KB Cells ; Oncogene Proteins ; metabolism

Objective To compare the pharmadynamics of fosfomycin in different dosing regimens to obtain the optimal clinical dosing regimen by Monte carlo simulation.Methods Pharmacokinetic datas of fosfomycin in lung tissue of septic patients were determined by in vivo microdialysis , application software ( Crystal Ball 7.2.2 ) performed Monte Carlo simula-tion of 10 000 patients to calculate the probability of target attainment ( PTA) of different doseing regimens.And combined with maximum drug concentration/minimum inhibitory concentration ( cmax/MIC).Results Of all the dosing regimens , PTA(6 g/q6 h) is best, however, when MIC>20, the PTA<50%.The probability of cmax/MIC≥4:prolonged infu-sion therapy ( PIT) (6 g/q6 h)>PIT(4 g/q6 h)>optimized two-step infusion therapy (OTIT) (6 g/q6 h)>OTIT(4 g/q6 h).When MIC≤16 , the antimicrobial PTA is approximately 100%,while the bactericidal PTA reduces to 66.48%.Conclusion When MIC≤20 , the prolonged infusion therapy of 6 g/6 h is best; while MIC>20 , PTA<50%, with poor clinical results , the best choice was drug combination.

Objective:To investigate the role of group 3 innate lymphoid cells (ILC3) in skin wound healing, and to explore the underlying mechanisms.Methods:Twenty-four 5-week-old male C57BL/6 mice were randomly and equally allocated into 3 groups: the skin wound + ILC3 inhibitor group (referred to as ILC3 inhibitor group), the skin wound group, and the control group, with 8 mice in each group. Four days before the establishment of the wound model, mice in the ILC3 inhibitor group were intraperitoneally injected with 1 μg of ILC3 inhibitor every 2 days for a total of 2 doses, mice in the skin wound group were injected with an equal volume of physiological saline solution, and mice in the control group were fed normally. To establish a mouse skin wound model, a full-thickness circular incision with a diameter of 0.6 cm was made around the midpoint of the dorsal midline using a biopsy punch after the intraperitoneal injection of anesthetics, which was histologically confirmed to be a full-thickness injury. The size of the wounds was observed and recorded, photographs of the wounds were taken on days 0, 1, 3, 5, 7, and 9 after wounding, and corresponding wound healing rates were calculated. On day 9 after wounding, tissue samples were collected from the wound edges, and subjected to flow cytometry analysis to quantify ILC3 infiltrating around the skin wound, and hematoxylin and eosin (HE) staining was performed to assess the healing status of the skin wounds. Real-time quantitative polymerase chain reaction (qRT-PCR) was conducted to determine the mRNA expression of vitamin D receptor (VDR), Notch1, tumor necrosis factor-alpha (TNF-α), interleukin (IL) -17A, IL-17F, and IL-22 in the wound-edge tissues, and Western blot analysis to determine their protein expression. Statistical analysis was carried out by using one-way analysis of variance and t test. Results:On day 9 after wounding, the skin wound group showed an increased number of ILC3 in the wound-edge tissues (5.31% ± 1.47% vs. 3.10% ± 0.54%, P < 0.01), increased mRNA and protein expression of TNF-α, IL-22, IL-17A, and IL-17F (all P < 0.05), but decreased mRNA and protein expression of VDR (both P < 0.05) compared with the control group; the protein expression of Notch1 was significantly higher in the skin wound group than in the control group ( P < 0.05), but there was no significant difference in its mRNA expression between the two groups ( P > 0.05). On days 1, 3 and 5, the wound healing rates were significantly higher in the ILC3 inhibitor group (45.17% ± 9.90%, 61.58% ± 11.61%, 75.61% ± 9.12%, respectively) than in the skin wound group (25.87% ± 10.96%, 47.78% ± 13.81%, 64.55% ± 10.29%, respectively, all P < 0.05). On day 9, the ILC3 inhibitor group showed a decreased number of ILC3 around the wound (2.69% ± 0.95%, P < 0.01), decreased mRNA and protein expression of TNF-α, IL-22, IL-17A, and IL-17F in the wound-edge tissues (all P < 0.05), but increased mRNA and protein expression of Notch1 and VDR in the wound-edge tissues (all P < 0.05) compared with the skin wound group. On day 9 after wounding, histopathological examination with HE staining revealed continuous and intact epithelial structure, as well as dense and neatly arranged collagen fibers in the ILC3 inhibitor group, and the structures of hair follicles, blood vessels, and sebaceous glands were similar to those in the control group. Conclusions:Skin ILC3 infiltrated local wounds and were involved in the skin wound healing process through inflammatory factors such as TNF-α, IL-17A, IL-17F, and IL-22. Downregulating the number of ILC3 may promote skin wound healing by activating VDR and Notch1, as well as inhibiting the TNF-α signaling pathway and the expression of downstream inflammatory factors.