Objective To evaluate the efficacy and safety of intravitreal ranibizumab injection combined with triamcinolone acetonide (TA) for macular edema secondary to type Ⅱ optic disc vasculitis.Methods Totally 19 cases(19 eyes) with macular edema secondary to type Ⅱ optic disc vasculitis from January 2013 to December 2015 were injected ranibizumab intravitreally and triamcinolone acetonide 2 weeks later.The visual acuity,intraocular pressure,ocular fundus,OCT,FFA and related complications were observed during the 6 months follow-up period.If the macuiar edema returns during the follow-up period,the eyes would be given ranibizumab and TA again until the macular edema subsided.Results The visual acuity was 0.16 ± 0.09 and the central retinal thickness was (694.88 ± 79.06) μm before treatment.Compared with pretreatment,the visual acuity of all cases were improved in different degree except for one at 1 month,3 months and 6 months (all P ＜ 0.05).Also central retinal thickness were decreased and macular edema were subsided,there were statistical differences (all P ＜0.05).Compared with results at 1 month,the visual acuity and central retinal thickness was no significant different at 3 months and 6 months (all P ＞ 0.05).The intraocular pressure of all cases were normal throughout the follow-up period but one was increased transitorily in the first day after intravitreal rauibizumab injection.FFA examination at 1 month showed retinal hemorrhage and effusion absorbed obviously,besides,the optic disc and macular fluorescein leakage reduced significantly.The macular edema of 1 eye returned at 3 months and subsided after giving ranibizumab and TA again.The average number of injected eyes was 2.11 times.No complications associated with intravitreal injection and drugs were observed during the follow-up period.Conclusion It is effective and safe that intravitreal ranibizumab junction combined with TA for macular edema secondary to type Ⅱ optic disc vasculitis,which not only shorten the period of treatment but also reduce the times of drug injection,surgical trauma and complications.

Objective To assess the prognosis of patients with bladder neck (BN) involvement in radical prostatectomy specimens and compare it with patients with seminal vesicle invasion (SVI).Methods From January 2002 to December 2008,42 patients with prostate carcinoma who underwent radical prostatectomy were analyzed retrospectively.The mean age was 70.3 years (range 59-78 years).The patients were divided into two groups according to 2002 TNM system:pT4a group (bladder neck invasion) with 17 cases and pT3b group (unilateral or bilateral seminal vesicle invasion)with 25 cases.Postoperative outpatients were followed-up on a regular basis,two consecutive postoperative serums PSA greater than 0.2 μg/L defined as a biochemical recurrence.The postoperative follow-up to the time of biochemical recurrence or follow up to the deadline without biochemical recurrence were defined as biochemical recurrence free survival time.Results All patients were followed up for 50-122 months.The patients with BN involvement 17.6％ (3/17) had biochemical recurrence,whereas seminal vesicle invasion 24.0％ (6/25),there was no statistical significance between the two groups (P＞0.05).Conclusions Postoperative adjuvant therapies can improve the biochemical recurrence free survival.In this study,the prognosis of pT4a and pT3b group is similar.A conceivable down-staging of BN involvement in the TNM staging system should be considered.

ObjectiveTo observe the clinical characteristic and treatment response of the Gleason 5 + 5 prostate cancer and to summarize the prognostic factor of such patients. MethodsFrom January 2005 to May 2010,21 cases of Gleason 5 + 5 prostate cancer were enrolled in this study.The average age was 73 years and the pre-treatment PSA was 60.8 ng/ml.Six of the 21 cases had a PSA level ≤20 ng/ml.The maximal androgen deprivation therapy (ADT) in the form of castration and Casodex or fluctamide was carried out once the diagnosis was made.Bone metastasis was revealed in 16 cases.PSA levels were tested at the 4th month and then every 3 months after the castration.The observation endpoint was 6 months.Extracorporeal beam irradiation or the systemic chemotherapy was carried out if the PSA did not drop to a normal range after 6 months. ResultsIn the study group with initial PSA ＞ 20 ng/ml,the PSA did not drop to normal range in 5 cases; of which 4 had bone metastasis.With systemic chemotherapy,3 cases died within 1 year.Another case,with localized disease,received an extracorporeal beam irradiation and died within 1 year due to progression of the tumor.Among the remaining 10 cases sensitive to total androgen blockage,7 survived through the end of the study period.One case had PSA recurrence in month seven after the initial therapy and died at month 19.One case had PSA recurrence at month 22 and died at month 36.The patient had his PSA augmented at month 24 after total androgen blockage and the patient passed away 4 months later.Of the 6 cases who had an initial PSA ≤20 ng/ml 4 died within one year. ConclusionsGleason 5 +5 prostate cancer is relatively resistant to ADT with a worse prognosis,especially for the cases with a nadir PSA ＞4 ng/ml after 6 months’ ADT.The patients who had an initial PSA ≤20 ng/ml might have an unsatisfactory clinical outcome.

OBJECTIVE: To observe the effect of thrombin on the cytotoxicity of astrocytes injured by hypoxia/reoxygenation(H/R) and to explore its relationship with inducible nitric oxide synthase (iNOS). METHODS: Primary astrocytes were cultured in DMEM with 10% approximately 15% calf serum and divided into 6 groups: a control group, a Tm control group, an H/R group, a Tm+H/R group, a hirudin (HR) control group, and a Tm+HR+ H/R group. The cell damage and viability were detected by the 3-(4, 5-di-methyl-thazol-2-yl)-2, 5 diphenyl-tetrazol-iumbromide (MTT) conversion method. The NO level in the cultured cell supernatant was assayed by Griess reagent. The flow cytometry was performed to evaluate the apoptosis rate of astrocytes. The iNOS mRNA was examined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Immunocytochemistry was used to observe the expression of iNOS protein. RESULTS: The cell viability injured by H/R was lower than that of the control group, the NO production and apoptosis rate in the cell of H/R group were higher than those of the control group. Incubation of H/R cell with 10kU/L Tm enhanced the cytotoxicity of H/R stimulation compared with the cells injured by H/R. Hirudin can reverse the effect of thrombin. RT-PCR and immunocytochemistry analysis demonstrated that the levels of iNOS mRNA and iNOS protein increased in the cells treated by H/R. Tm enhanced the expression of iNOS mRNA and iNOS protein in the cells treated by H/R. Hirudin blocked the effect of Tm. CONCLUSION Increasing the level of iNOS and enhancing the production of NO may be the mechanism of thrombin cytotoxicity in astrocytes injured by H/R.
Animals ; Apoptosis ; drug effects ; Astrocytes ; cytology ; drug effects ; Cell Hypoxia ; Cells, Cultured ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Rats ; Rats, Sprague-Dawley ; Thrombin ; pharmacology

OBJECTIVETo study the protective effects of polysaccharide of Spirulina platensis and Sargassum thunbeergii on vascular of alloxan (ALX) induced diabetic rats.

METHODWith the doses of polysaccharide of Spirulina platensis (PSP) and Sargassum thunbeergii (PST) compound (1:1) 12.261, 36.783, 110.349 mg x kg(-1) by i.g. administration to alloxan induced diabetic rats respectively for 6 weeks. Then the blood glucose and the TC, HDL-C, TG, NO, ET in serum were detected. The contraction and relaxation response to NE and ACh in aortic rings of the alloxan induced diabetic rats has been studied.

RESULTThe results showed the compound of PSP and PST could decrease the blood glucose and the TC, TG, NO, ET in serum and increase HDL-C than in the alloxan induced diabetic rats. The contraction responses to NE in aortic rings of the alloxan induced diabetic rats were significantly elevated in the normal rats, and the responses to ACh were significantly lower. PSP and PST compound could significantly lower the responses to NE and significantly elevate the responses to ACh in aortic rings of the alloxan induced diabetic rats.

CONCLUSIONPSP and PST compound could decrease blood glucose and could protect the vascular of alloxan induced diabetic rats.

Animals ; Aorta, Thoracic ; drug effects ; Blood Glucose ; metabolism ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cyanobacteria ; chemistry ; Diabetes Mellitus, Experimental ; blood ; Endothelins ; blood ; Female ; Male ; Mice ; Muscle Contraction ; drug effects ; Muscle, Smooth, Vascular ; drug effects ; Nitric Oxide ; blood ; Polysaccharides ; isolation & purification ; pharmacology ; Polysaccharides, Bacterial ; isolation & purification ; pharmacology ; Protective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sargassum ; chemistry ; Triglycerides ; blood

OBJECTIVETo investigate the effect of poly-ADP-ribosylation in hexavalent chromium Cr(VI) induced cell damage.

METHODSThe study object, poly (ADP-ribose) glycohydrolase (PARG) deficient human bronchial epithelial cells (16HBE cells), was constructed previously by our research group. Normal 16HBE cells and PARG-deficient cells were treated with different doses of Cr (VI) for 24 h to compare the differences to Cr (VI) toxicity, meanwhile set up the solvent control group. On this basis, 5.0 µmol/L of Cr (VI) was selected as the exposure dose, after the exposure treatment, total proteins of both cells were extracted for two dimension fluorescence difference gel electrophoresis (2D-DIGE) separation, statistically significant differential protein spots were screened and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS/MS), and further validated by Western blot.

RESULTSAfter Cr (VI) treatment, the survival rate of PARG-deficient cells was higher than normal 16HBE cells. When the doses reached up to 5.0 µmol/L, the survival rate of 16HBE cells and PARG-deficient cells were respectively (59.67 ± 6.43)% and (82.00 ± 6.25)%, the difference between which was significant (t = -4.32, P < 0.05). 18 protein spots were selected and successfully identified after 2D-DIGE comparison of differential proteins between normal 16HBE cells and PARG-deficient cells before and after exposure. The function of those proteins was involved in the maintenance of cell shape, energy metabolism, DNA damage repair and regulation of gene expression. The differential expression of cofilin-1 was successfully validated by Western blot. The expression level of cofilin-1 in the 16HBE cells increased after Cr (VI) exposure with the relative expression quantity of 1.41 ± 0.04 in treated group and 1.00 ± 0.01 in control group, the difference of which was statistically significant (t = -18.00, P < 0.05), while the expression level in PARG-deficient cells had no statistically significant difference (t = -8.61, P > 0.05).

CONCLUSIONMost of the identified differential proteins are closely related to tumorigenesis, suggesting that poly-ADP-ribosylation reaction may resist the cytotoxicity of Cr(VI) by inhibiting Cr (VI) induced tumorigenesis, which provides important reference data to clarify the mechanisms of poly-ADP-ribosylation in Cr (VI) induced cell damage.

Bronchi ; Cell Transformation, Neoplastic ; genetics ; Chromium ; Cofilin 1 ; DNA Repair ; Epithelial Cells ; Glycoside Hydrolases ; deficiency ; physiology ; Humans ; Tandem Mass Spectrometry

OBJECTIVETo explore the effects of pre-acupuncture and immediate acupuncture on kidney function and oxygen free radical metabolism in rats with simulated weightlessness.

METHODSTwenty male clean-grade Wister rats were randomly divided into a normal control group, a model group, a pre-acupuncture group and an immediate acupuncture group, 5 rats in each one. The rats in the normal control group did not receive any treatment but free activities for 4 weeks. The rats in the rest groups received 4-week tail suspension to establish the model of simulated weightlessness. One week before the tail suspension, the rats in the pre-acupuncture group were treated with electroacupuncture at "Shenshu" (BL 23), "Pishu" (BL 20) and "Sanyinjiao" (SP 6) for 30 min per treatment, once a day for 7 days. The rats in the immediate acupuncture group received tail suspension and acupuncture at the same time; during the tail suspension, the electroacupuncture was applied at "Shenshu" (BL 23), "Pishu" (BL 20) and "Sanyinjiao" (SP 6) for 30 min per treatment, once every other day for 14 days. The colorimetric method was used to measure the content of blood urea nitrogen (BUN) in serum as well as activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) and content of malonaldehyde (MDA) in renal tissue in each group.

RESULTSCompared with the normal control group, the content of BUN in the model group was increased significantly (P<0.01), the activity of SOD and GSH-PX in nephridial tissue was significantly reduced (both P<0.01), and the content of MDA was increased significantly (P<0.05). Compared with the model group, the content of BUN in the pre-acupuncture group and immediate acupuncture group was significantly reduced (P<0.01, P<0.05), the activity of GSH-PX in the pre-acupuncture group was obviously increased (P<0.05) and the content of MDA in the immediate acupuncture group was increased significantly (P<0.01). Compared with the immediate acupuncture group, the content of MDA in the pre-acupuncture group was lower (P<0.01).

CONCLUSIONThe pre-acupuncture and immediate acupuncture both have the capacity to improve the kidney function and anti-oxygen free radical injury in rats with simulated weightlessness, however, the capacity to increase the protection ability of the kidney and eliminate free radical in the pre-acupuncture group is superior to that in the immediate acupuncture group, which is likely to be related with improving antioxidant ability of kidney.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Antioxidants ; metabolism ; Humans ; Kidney ; metabolism ; Kidney Diseases ; metabolism ; physiopathology ; therapy ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Space Flight ; Superoxide Dismutase ; metabolism ; Weightlessness ; adverse effects

Objective To evaluate a model for axillary lymph node involvement combining CK19 mRNA with contrast enhanced ultrasound sonography (CEUS) score in operable breast cancer.Methods Operable breast cancer patients planned for sentinel lymph node (SLN) biopsy were enrolled.Preoperative CK19mRNA expressions in peripheral blood and CEUS score of axillary lymph nodes were tested before surgery.In the training set,postoperative sentinel lymph node (SLN) and non-sentinel lymph node (nSLN) pathological results were taken as the gold standard,effective modeling variables were screened,logistic regression was used to establish the prediction model.Parallel control studies were conducted between the validation set and the MSKCC model to evaluate the prediction accuracy and prediction efficiency.Results From Oct 2015 to Nov 2016,359 cases (training set) were enrolled and mathematical formulas for predicting SLN and nSLN were established,respectively.The sensitivity,specificity and AUC of predicting SLN involvement were 91.36％,94.92％ and 0.979 respectively.The sensitivity,specificity and AUC of predicting nSLN metastasis were 91.04％,90.53％ and 0.932 respectively.From Dec 2016 to Jul 2017,219 cases (verification set) were included.The sensitivity of SLN metastasis predicted by the model was 91.84％,the specificity was 96.69％,and the AUC was 0.979,significantly superior to the MSKCC model (0.739).The sensitivity,specificity and AUC of predicting nSLN metastasis were 95.35％,92.73％ and 0.945 respectively,significantly superior to the MSKCC model (0.873).Concolusions Combined with peripheral blood CK19 mRNA and CEUS score,the prediction model for axillary lymph node involvement for operable breast cancer,SLN/nSLN involvement probability can be calculated and qualitative judgment can be made.The overall accuracy and AUC of this model are better than the prediction model of MSKCC.

Objective To construct a RNA interference lentiviral vector aimed at human ARK5 (AMPK-related protein kinase 5) gene and explore its effect on the biologic behavior of human gastric cancer SGC7901 cells.Metho ds Targeting human ARK5 mRNA coding sequence, we designed three specific short hairpin RNAs (shRNAs) and constructed the lentiviral vector,then infected human gastric cancer SGC7901 cells with this vector.Afterwards, we used qPCR and Western blot for detecting the silencing effect on ARK5 gene,MTT colorimetric assay to measure the cell proliferation , cell scratch test for cell migration and Transwell for cell invasion, and flow cytometry analysis for apoptosis in cells treated with glucose starvation and TNF-α.Results Sequencing proved that the recombinant lentiviral vector containing ARK5-shRNA-3 was constructed successfully.Real time fluorescent quantitative PCR assay showed that the expression abundance of ARK5 gene in the normal control group, negative control group and ARK5-shRNA-3 infected group were 1.002+0.082, 1.001+0.050 and 0.140+0.003, respectively, showing a statistically significant difference (P<0 .01).Cell scratch test showed that the cell migration rate of ARK5-shRNA-3 infected group was (38.5+4.3 )%, significantly lower than that of the normal control group [(72.4+6.4)%] and negative control group [(75.1+7.1)%, P<0.01].The results of Transwell test showed that the number of penetrating cells in the normal control group, negative control group and ARK5-shRNA-3 transfection group were 257.4±12.3, 245.7±11.6, 112.5±7.8, with a significant difference (P<0.01).After glucose starvation and TNF-α-treatment for 24 h, the cell death rate of the normal control group, negative control group and ARK5-shRNA-3 group were (11.7±3.2)%, (12.3±2.6)% and (30.8±4.3)%, respectively, showing that the cell apoptosis rate of ARK5-shRNA-3 transfected group was significantly higher than that of the normal control and negative control groups (P<0.01).Conclusions We have successfully constructed a recombinant lentiviral vector which can efficiently silence ARK5 gene.Using it we can inhibit the proliferation, migration, invasion of tumor cells, and promote cell apoptosis under the condition of TNF-αtreatment and glucose starvation.

Objective To construct a RNA interference lentiviral vector aimed at human ARK5 (AMPK-related protein kinase 5) gene and explore its effect on the biologic behavior of human gastric cancer SGC7901 cells.Metho ds Targeting human ARK5 mRNA coding sequence, we designed three specific short hairpin RNAs (shRNAs) and constructed the lentiviral vector,then infected human gastric cancer SGC7901 cells with this vector.Afterwards, we used qPCR and Western blot for detecting the silencing effect on ARK5 gene,MTT colorimetric assay to measure the cell proliferation , cell scratch test for cell migration and Transwell for cell invasion, and flow cytometry analysis for apoptosis in cells treated with glucose starvation and TNF-α.Results Sequencing proved that the recombinant lentiviral vector containing ARK5-shRNA-3 was constructed successfully.Real time fluorescent quantitative PCR assay showed that the expression abundance of ARK5 gene in the normal control group, negative control group and ARK5-shRNA-3 infected group were 1.002+0.082, 1.001+0.050 and 0.140+0.003, respectively, showing a statistically significant difference (P<0 .01).Cell scratch test showed that the cell migration rate of ARK5-shRNA-3 infected group was (38.5+4.3 )%, significantly lower than that of the normal control group [(72.4+6.4)%] and negative control group [(75.1+7.1)%, P<0.01].The results of Transwell test showed that the number of penetrating cells in the normal control group, negative control group and ARK5-shRNA-3 transfection group were 257.4±12.3, 245.7±11.6, 112.5±7.8, with a significant difference (P<0.01).After glucose starvation and TNF-α-treatment for 24 h, the cell death rate of the normal control group, negative control group and ARK5-shRNA-3 group were (11.7±3.2)%, (12.3±2.6)% and (30.8±4.3)%, respectively, showing that the cell apoptosis rate of ARK5-shRNA-3 transfected group was significantly higher than that of the normal control and negative control groups (P<0.01).Conclusions We have successfully constructed a recombinant lentiviral vector which can efficiently silence ARK5 gene.Using it we can inhibit the proliferation, migration, invasion of tumor cells, and promote cell apoptosis under the condition of TNF-αtreatment and glucose starvation.