BACKGROUNDCataracts is considered be formed because of an abnormal glucose metabolic pathway or oxidative stress. We explored the damaging role of ONOO- and antagonism of cholecystokinin octapeptide-8 (CCK-8) in diabetic cataractal rat lenses.

METHODSA diabetic cataractal animal model was established by peritoneal injection of streptozotocine (STZ). Thirty-six normal SD rats were taken as control group; seventy-two were given STZ (45 mg/kg) and then divided into STZ group and CCK-8 group (peritoneal injection CCK-8). STZ induced diabetic rats were treated with CCK-8 for 60 days. Lenses were examined with slit lamp at 20, 40 and 60 days. Immunofluorescent staining and Western blot analysis were used for determining nitrotyrosine (NT, a marker for ONOO-). PT-PCR and gene array analysis were used for determining the expression of inducible nitric oxide synthetase mRNA (iNOS mRNA) in lens epithelium (LEC).

RESULTSSTZ group rats developed lens opacity by 20 days that reached a high level by 60 days after STZ injection. CCK-8 group rats delayed the cataract formation. CCK-8 group rats delayed the cataract formation. There was no distinct expression of NT and iNOS mRNA in control group. In STZ group, there were distinct expression of NT and upregulation of iNOS mRNA; however, CCK-8 group showed weak expression of NT and downregulation of iNOS mRNA.

CONCLUSIONSNT, which may be a new form of oxidative stress, was expressed in diabetic rat LEC although CCK-8 could reverse NT damage in LEC. The results suggested that CCK-8 might be a useful therapeutic agent against diabetic cataract. The antagonizing mechanism of CCK-8 may be related to direct antagonism of ONOO- as well as its inhibition of the expression of iNOS mRNA for production of NO and therefore decrease in the formation of ONOO-.

Animals ; Blotting, Western ; Cataract ; etiology ; prevention & control ; Diabetes Mellitus, Experimental ; complications ; Fluorescent Antibody Technique ; Male ; Nitric Oxide Synthase Type II ; genetics ; Oxidation-Reduction ; Peroxynitrous Acid ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Sincalide ; pharmacology ; Streptozocin ; Tyrosine ; analogs & derivatives ; genetics

The present study was designed to observe if puerarin decreases lens epithelium cell (LEC) apoptosis induced partly by peroxynitrite (ONOO(-)). One hundred and eight rats were randomly divided into control group (n=36), streptozotocin (STZ) group (n=36) and STZ + puerarin group (n=36). The rats in the control group intraperitoneally (i.p.) received 0.5 ml of saline. The rats in STZ group and STZ + puerarin group received intraperitoneal injection of STZ (45 mg/kg). Three days later, the rats in STZ + puerarin group were given puerarin (140 mg/kg per day, i.p.). On days 20, 40 and 60 of the experiment, morphologic changes of lenses were observed with slit lamp. Then the animals were sacrificed for further analysis. The amount and percentage of apoptotic LECs were determined by flow cytometry. Nitrotyrosine (NT, the foot print of ONOO(-)) was examined by immunohistochemistry. Apoptosis-related genes (iNOS, etc.) were analyzed by gene array. The results showed that in the control group, all the lenses were clear. In STZ group, gradually severe opacity of the lens was observed on days 20, 40 and 60. But in STZ + puerarin group, mild opacity of the lens was observed on day 20 and more severe on day 40, but markedly decreased on day 60. In the control group, mild apoptosis of LECs was observed. In STZ group, time-dependent increase in apoptosis of LECs was observed. In STZ + puerarin group, mild apoptosis of LECs was observed on day 20, significantly increased on day 40, but markedly decreased on day 60. There was no expression of NT in the lens in the control group, but an increased expression of NT in STZ group. In STZ + puerarin group, mild expression of NT was observed on day 20, significantly increased on day 40, but markedly decreased on day 60. There was no expression of iNOS in the lens in the control group, but continuous up-regulation of iNOS expression in STZ group. In STZ + puerarin group, mild expression of iNOS was observed on day 20, significantly increased on day 40, but markedly decreased on day 60. Except the changes of iNOS related to NO production, the other apoptosis-related genes, including BCL-2 and SOD were down-regulated, while NF-kappaB and TNFR1-FADD-caspase signal transduction way were up-regulated in STZ group. The results were opposite in STZ + puerarin group and the control group. These findings show that NT is expressed in diabetic rat lens, which proves that LEC apoptosis in diabetic lens is partly induced by ONOO(-) which may be a new oxidative damage way to form cataract. Puerarin partly decreases LEC apoptosis induced by ONOO(-) and is a potential medicine for therapy of diabetic cataract. The mechanism of puerarin dealing with diabetic cataract may be related to its direct inhibition of LEC apoptosis and antagonism of ONOO(-) in diabetic rats.
Animals ; Apoptosis ; Cataract ; chemically induced ; Diabetes Mellitus, Experimental ; Epithelial Cells ; drug effects ; Isoflavones ; pharmacology ; Lens, Crystalline ; cytology ; Nitric Oxide Synthase Type II ; metabolism ; Peroxynitrous Acid ; Rats ; Tyrosine ; analogs & derivatives ; metabolism

OBJECTIVETo investigate effect of inhibiting melatonin biosynthesis on activities of protein kinase A (PKA), glycogen synthase kinase-3 (GSK-3) and tau phosphorylation at PS214 and M4 epitopes using haloperidol, a specific inhibitor of 5-hydroxyindole-O-methyltransferase.

METHODSBrain ventricular and intraperitoneal injections were used for haloperidol administration, Western blots for tau phosphorylation, 32P-labeling for PKA and GSK-3 activity, and high performance liquid chromatograph for detection of serum melatonin levels.

RESULTSHaloperidol injection through the lateral ventricle and intraperitoneal reinforcement significantly stimulated PKA activity with a concurrent hyperphosphorylation of tau at M4 (Thr231/Ser235) and PS214 (Ser214) sites. Prior treatment of the rats using melatonin supplement for one week and reinforcement during the haloperidol administration arrested PKA activity and attenuated tau hyperphosphorylation. GSK-3 activity showed no obvious change after haloperidol injection, however, melatonin supplements and reinforcements during haloperidol infusion inactivated basal activity of GSK-3.

CONCLUSIONDecreased melatonin may be involved in Alzheimer-like tau hyperphosphorylation, and overactivation of PKA may play a crucial role in this process.

Animals ; Cyclic AMP-Dependent Protein Kinases ; metabolism ; Epitopes ; Glycogen Synthase Kinase 3 ; metabolism ; Haloperidol ; administration & dosage ; pharmacology ; Hippocampus ; enzymology ; metabolism ; Injections, Intraperitoneal ; Injections, Intraventricular ; Male ; Melatonin ; biosynthesis ; blood ; Phosphorylation ; drug effects ; Rats ; Rats, Wistar ; tau Proteins ; metabolism

Background: In the era of intensity-modulated radiotherapy (IMRT), the role of neoadjuvant chemotherapy (NACT) in treating ascending-type nasopharyngeal carcinoma (NPC) is under-evaluated. This study was to compare the efficacy of NACT followed by IMRT (NACT + RT) with the efficacy of concurrent chemoradiotherapy (CCRT) on ascending-type NPC.Methods: Clinical data of 214 patients with ascending-type NPC treated with NACT + RT or CCRT between December 2009 and July 2011 were analyzed. Of the 214 patients, 98 were treated with NACT followed by IMRT, and 116 were treated with CCRT. The survival rates were assessed using Kaplan –Meier analysis, and the survival curves were compared using a log-rank test.Results: The 4-year overall survival, locoregional failure-free survival, distant failure-free survival, and failure-free survivalrates were not significantly different between the two groups (all P > 0.05). However, patients in the CCRT group exhibited more severe acute adverse events than did patients in the NACT + RT group during radiotherapy, including leukopenia (30.2% vs. 15.3%, P = 0.016), neutropenia (25.9% vs. 11.2%, P = 0.011), and mucositis (57.8% vs. 40.8%, P = 0.028). After radiotherapy, patients in the CCRT group exhibited significantly higher rates of xerostomia (21.6% vs.10.2%, P = 0.041) and hearing loss (17.2% vs. 6.1%, P = 0.023).Conclusions: The treatment outcomes of the NACT + RT and CCRT groups were similar; however, CCRT led to higher rates of acute and late toxicities. NACT + RT may therefore be a better treatment strategy for ascending-type NPC.

Objective To analysis the effect of different age factor on the prognosis and efficacy of acute cerebral infraction in patients treated by intravenous thrombolytic therapy with alteplase.Methods Ninety-eight patients with cerebral infarction were divided into older group (n=52) and younger group(n=46).Patients in two groups were trea-ted by intravenous thrombolysis with 0.9 mg · kg-1 alteplase.After 1 and 21 d, the efficacy was observed and scored using national institute of health stroke scale( NIHSS).At the same time,the incidence of adverse drug reactions were observed, and prognosis evaluated using Modified Rankin scale ( mRS ) in 3 weeks.Results NIHSS scores were signifi-cantly lower in the both groups treated by alteplase than those before thera-py( P<0.05).In 1 and 21 d, NIHSS scores were lower in the younger groups than those in elderly group ( P<0.05 ).The incident of adverse drug reaction was lower in the younger groups than the elderly group ( P<0.05).It has shown that the prognosis was better in the younger group than the elderly group ( P<0.05 ) by comparing mRS score in 3 weeks.Conclusion All acute cerebral infraction patients received sig-nificant benefits from the intravenous thrombolytic therapy with alteplase.The younger group had more significant efficacy, less complications and better prognosis than those of elderly group.

OBJECTIVETo investigate the occurrence of important foodborne pathogens in shellstock Pacific oysters in the food markets in South China.

METHODSFrom July 2007 to June 2008, retail oysters were collected in different seasons from South China and analyzed for the prevalence and levels of Listeria monocytogenes, Vibrio vulnificus and Vibrio parahaemolyticus.

RESULTSNone of L. monocytogenes could be detected in any of the 202 oyster samples tested, while E vulnificus and V. parahaemolyticus could be detected in 67 (54.9%) and 109 (89.3%) of the 122 oyster samples analyzed, respectively, with an MPN (most probable number) value greater than or equal to 3. V. vulnificus and V. parahaemolyticus with a more than 102 MPN/g were found in 36 (29.5%) and 59 (48.4%) of the 122 oyster samples, respectively. The tdh and trh genes were detected in 4 (0.3%) and 8 (0.6%) of the 1 349 V parahaemolyticus isolates, respectively. Of the 122 samples, 4 (3.3%) was positive for either tdh or trh. The levels of V. vulnificus and total V. parahaemolyticus in oysters in South China varied in different seasons.

CONCLUSIONV. vulnificus and pathogenic V. parahaemolyticus are frequently found in oysters in south China, which may pose a potential threat to public health. Data presented here will be useful for the microbiological risk assessment in oysters in China.

Animals ; China ; Commerce ; Food Microbiology ; Listeria monocytogenes ; isolation & purification ; Ostreidae ; microbiology ; Vibrio parahaemolyticus ; isolation & purification ; Vibrio vulnificus ; isolation & purification

OBJECTIVETo investigate the anti-angiogenic effects of Pien Tze Huang in vivo and in vitro.

METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with 0 mg/mL, 0.25 mg/mL, 0.5 mg/mL, and 1 mg/mL of PZH for 24 h, 48 h and 72 h, respectively. Chicken embryo chorioallantoic membrane (CAM) model was used to evaluate in vivo angiogenesis. An ECMatrix gel system was used to evaluate in vitro angiogenesis by examining the tube formation of HUVECs. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine HUVEC viability. Cell density of HUVECs was observed by phase-contrast microscopy. HUVEC migration was determined by wound healing method. The mRNA and protein expression of vascular endothelial growth factor A (VEGF-A) and basic fibroblast growth factor (bFGF) in both HUVEC and human colon adenocarcinoma cells (HT-29) was examined by reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immune sorbent assay (ELISA), respectively.

RESULTSPZH treatment significantly reduced the total number of blood vessels compared with the untreated control in the chicken embryos and resulted in a significant decrease in capillary tube formation and cell density of HUVECs (P<0.05). In addition, treatment with 0.25-1 mg/mL of PZH for 24 h, 48 h, and 72 h respectively reduced cell viability by 9%-52%, 24%-87% or 25%-87%, compared with the untreated control cells (P<0.05). Moreover, PZH treatment decreased the migration of HUVECs. Furthermore, PZH dose-dependently suppressed the expression of VEGF-A and bFGF on both mRNA and protein levels (P<0.05).

CONCLUSIONPZH could inhibit angiogenesis in vivo in CAM model and in vitro on HUVECs, suggesting that inhibiting tumor angiogenesis might be one of the mechanisms by which PZH treats cancer.

Animals ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fibroblast Growth Factor 2 ; genetics ; metabolism ; Gene Expression Regulation ; drug effects ; HT29 Cells ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Neovascularization, Physiologic ; drug effects ; genetics ; RNA, Messenger ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo evaluate hemodynamic character and surgical effect of stenoses or occlusion of carotid artery system by perfusion-weighted MRI.

METHODSTwenty-one patients with stenoses or occlusion of carotid artery system underwent surgical treatment. Two patients underwent endarterectomy, extracranial-intracranial arterial bypass to 10 patients, and percutaneous stent angioplasty to 9 patients.

RESULTSPreoperative perfusion-weighted magnetic resonance (MR) revealed normal regional cerebral blood flow in all patients, with delayed time to peak in both middle cerebral artery distribution and border zone. Postoperative perfusion-weighted MR revealed normal time to peak in border zone, but abnormal in middle cerebral artery distribution. Evaluated by perfusion-weighted MR, it showed that surgical method can improve the hemodynamic disorder of this kind of disease. The early curative effect of endarterectomy and percutaneous stent angioplasty is better than extracranial-intracranial arterial bypass.

CONCLUSIONSPerfusion-weighted MR is a good method to evaluate hemodynamic character of stenoses or occlusion of carotid artery system. Surgical method is helpful to this kind of disease, and its curative effect can be evaluated impersonally and accurately by this new technique.

Adult ; Aged ; Angioplasty ; Carotid Stenosis ; physiopathology ; surgery ; Cerebrovascular Circulation ; Endarterectomy, Carotid ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Angiography ; methods ; Male ; Middle Aged ; Sensitivity and Specificity ; Treatment Outcome

BACKGROUNDCarotid artery stenting (CAS) as a competing treatment modality has had to adhere to limits to gain widespread acceptance in some studies. This study analyzed the clinical data of 1700 consecutive patients after CAS to retrospectively evaluate the 30-day outcome of CAS for internal carotid artery stenosis in a Chinese population.

METHODSMedical records of 1700 patients who underwent CAS at Xuanwu Hospital affiliated to Capital Medical University between January 2001 and August 2012 were reviewed. Postoperative 30-day complication rates were analyzed and compared with those of other studies. Univariate and multivariate Logistic regression analyses were used to identify factors associated with perioperation myocardial infarction (MI), stroke, and death.

RESULTSThe overall 30-day rate of MI, stroke, and death after CAS was 2.53%. In univariate analysis, patients who were symptomatic, had a neurological deficit (modified Rankin score (mRS) ≥3; P = 0.001), and who were not taking statins experienced a significantly increased rate of MI, stroke, and death (P = 0.017). In multivariate Logistic regression analysis, the presence of symptoms (odds ratio (OR) = 2.485; 95% confidence interval (CI) = 1.267-4.876; P = 0.008) and a neurological deficit (mRS ≥3) (OR = 3.025; 95% CI = 1.353-6.763; P = 0.007) were independent risk factors for perioperative MI, stroke, and death.

CONCLUSIONSAccording to this single-center experience, CAS may effectively prevent and treat carotid artery stenosis that would otherwise lead to stroke. Being symptomatic and having a neurological deficit (mRS ≥3) increased the risk of perioperative MI, stroke, and death.

Adult ; Aged ; Aged, 80 and over ; Carotid Stenosis ; surgery ; Female ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Myocardial Infarction ; Stents ; Stroke ; surgery ; Treatment Outcome

Clinical target volume (CTV) delineation is crucial for tumor control and normal tissue protection. This study aimed to define the locoregional extension patterns of nasopharyngeal carcinoma (NPC) and to improve CTV delineation. Magnetic resonance imaging scans of 2366 newly diagnosed NPC patients were reviewed. According to incidence rates of tumor invasion, the anatomic sites surrounding the nasopharynx were classified into high-risk (>30%), medium-risk (5%-30%), and low-risk (<5%) groups. The lymph node (LN) level was determined according to the Radiation Therapy Oncology Group guidelines, which were further categorized into the upper neck (retropharyngeal region and level II), middle neck (levels III and Va), and lower neck (levels IV and Vb and the supraclavicular fossa). The high-risk anatomic sites were adjacent to the nasopharynx, whereas those at medium-or low-risk were separated from the nasopharynx. If the high-risk anatomic sites were involved, the rates of tumor invasion into the adjacent medium-risk sites increased; if not, the rates were significantly lower (P<0.01). Among the 1920 (81.1%) patients with positive LN, the incidence rates of LN metastasis in the upper, middle, and lower neck were 99.6%, 30.2%, and 7.2%, respectively, and skip metastasis happened in only 1.2% of patients. In the 929 patients who had unilateral upper neck involvement, the rates of contralateral middle neck and lower neck involvement were 1.8% and 0.4%, respectively. Thus, local disease spreads stepwise from proximal sites to distal sites, and LN metastasis spreads from the upper neck to the lower neck. Individualized CTV delineation for NPC may be feasible.
Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; pathology ; Neck ; pathology ; Neoplasm Invasiveness ; Tumor Burden