ObjectiveTo study the effect of testing amino acids levels in brain microdialysate by method of high performance liquid chromatography (HPLC) with α-aminobutyric acid (AABA) as the internal standard.MethodsReversed phase high performance liquid chromatography (RP-HPLC), orthophthaladehyde (OPA) precolumn derivatization method and fluorescence detector were used. The separation was performed using an ODS-C18 column. AABA, an isomeric compound of GABA, served as the internal standard, phosphate buffer solution and methanol as the mobile phase, gradient elution lasted in 35 min.Results7 kinds of amino acids and internal standard were separated completely. The mean recovery of amino acid was 88.2%—102.3%. The mean correlation coeffeicient of the linear relationship was 0.998±0.0015.Limits of detection for amino acids were 1.0—8.6 ppm respectively.ConclusionThe method mentioned above is simple and sensitive for operation, and can be used for determined levels of amino acids in microdialysate.

The effects of endothelin-1 (ET-1) and other drugs on the reactive oxygen species (ROS) generation and cardiomyocyte hypertrophy were examined in experiments on the cultured neonatal rat cardiomyocytes. The role of ROS on neonatal rat cardiomyocyte hypertrophy induced by ET-1 was studied and the relationship of PKC activation and ROS generation was investigated. The level of intracellular ROS was measured by the ROS-specific probe 2',7'-dichlorofluorescin diacetate (DCF-DA). Cardiomyocyte hypertrophy was determined by the RNA content, the total protein of cells and the cell surface area. The results are as follows. The fluorescence intensity of intracellular DCF-DA increased by 77% in cultured neonatal rat cardiac myocytes treated with ET-1 (10 nmol/L) vs control group. Compared with control group, the fluorescence intensity of intracellular PI, protein content and cell surface area increased by 128%, 87% and 151% respectively (all P<0.01) in cardiac myocytes treated with ET-1 (10 nmol/L). ABT-627, CC, or CAT inhibited the ET-1-induced increase in fluorescence intensity of intracellular DCF-DA by 62%,60% and 51% respectively (all P<0.01), and also attenuated the cardiac hypertrophy. The fluorescence intensity of intracellular DCF-DA increased by 74% (P<0.01) in myocytes treated with PMA (1 micromol/L) vs control group. Therefore, in the course of cardiomyocyte hypertrophy, ET-1 increases intracellular ROS in the cultured neonatal rat cardiac myocytes and inhibits cardiomyocyte hypertrophy induced by ROS. The ET(A) and PKC activation mediate the ROS production and cardiomyocyte hypertrophy induced by ET-1. ROS is necessary in the ET-1-induced cardiomyocyte hypertrophy.
Animals ; Animals, Newborn ; Cell Enlargement ; Cells, Cultured ; Endothelin-1 ; physiology ; Female ; Hypertrophy ; Male ; Myocytes, Cardiac ; cytology ; pathology ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism

Objective To investigate the expressions of nerve growth factor(NGF)/TrkA in pancreatic cancer and evaluate their association with perineural invasion and pain in patients with pancreatic cancer. Methods Twenty-eight patients with pancreatic cancer were divided into group A wim pain(n=14)and group B without pain(n=14),and in all the patients the expressions of NGF/TrkA in the pancreatic CanCel tissue were detected by Northern blot analysis and immunohistochemistry.The molecular findings were analyzed in relation to the perineural invasion scores and pain scores.Results Perineural invasion scores in group A were significantly higher than that in group B (P＜0.05).Northern blot analysis revealed significantly higher NGF/TrkA levels and NGF/TrkA immunoreactivity in group A than in group B (P＜0.05).Immunohistochemistry showed that NGF was strongly expressedin the cytoplasm of the pancreatic cancer cells but TrkA Was localized in the perineurinm of the pancreatic Berve.NGF and TrkA mRNA expressions in the tumors were found to positively correlate to perineural invasion of the tumors(r=622,P＜0.05,and r=0.681,P＜0.05,respectively)and the pain scores(r=0.624,P＜0.05;r=0.632,P＜0.05).The immunohistochemical scores of NGF and TrkA expressions in the tumor were also positively correlated to perineural invasion of the tunlor(r=0.602,P＜0.05;r=0.551,P＜0.05)and the pain scores(r=q).603,P＜0.01;r=0.612,P＜0.01).Conclusions The NGF/TrkA system might play important roles in nerve infiltration by the tumor cells and pain sensation in pancreatic cancer patients.

Hereditary spastic paraplegia(HSP or SPG) is a clinically and genetically heterogeneous group of neurodegenerative diseases characterized by progressive spasticity, weakness of lower limbs, and pathologically by retrograde axonal degeneration of corticospinal tracts and posterior spinal tracts. Presence of additional features allows differentiation between simple and complex forms of the disease. Genetically, 16 loci for HSP accompanied by distal amyotrophy have been mapped, for which 13 genes have been identified. With the identification of causative genes, the molecular mechanism of this disease is gradually elucidated.
Brachial Plexus Neuritis ; complications ; genetics ; Genetic Heterogeneity ; Humans ; Spastic Paraplegia, Hereditary ; complications ; genetics

Objective This study was to evaluate the clinical outcomes of orbital blowout fracture repair using the individual titanium mesh bent by computer-aided rapid prototyping . Methods The CT imaging data of 11 cases of orbital blowout fracture were analyzed.The subciliary approach was used for the exposure of the fractures .An appropriate 3D-printed titanium mesh pre-bent by computer-aided rapid prototyping was selected and implanted according to the characteristics of a given defect .The surgical results were analyzed by evaluating diplopia , eyeball movement , enophthalmos , and the position of the titanium mesh and comparing the orbital vol-ume before and after surgery . Results All the operations were successfully accomplished .No remolding or trimming was needed for the pre-bent titanium mesh and implanted titanium mesh was tightly fixed to the orbital wall .The patients were followed up for 3 -12 months, which revealed no symptoms of diplopia in any of the patients . Volumetric analysis showed a significantly reduced orbital volume difference from (2.60 ±0.43) mL preoperatively to (-0.07 ±0.62) mL postoperatively (P<0.01).Enophthalmos was desirably correc-ted, with the enophthalmos difference decreased from (2.35 ±0.81)mm preoperatively to (-0.10 ±0.52) mm postoperatively (P<0.01).No extra-ocular muscle limitation was observed . Conclusion The individual titanium mesh bent with computer-aided rapid prototyping technology can be applied to orbital blowout fracture repair , which may achieve a high success rate of anatomic restoration of the orbital volume and effective prevention of enophthalmos .

OBJECTIVETo investigate the significance of monitoring procalcitonin (PCT) when applying antibiotics to trichlorethylene (TCE)-induced dermatitis.

METHODSOne hundred and two patients who were hospitalized and recovered from TCE-induced dermatitis in our hospital from 2006 to 2013 were enrolled as subjects. Based on whether the PCT level was monitored or not, we divided patients into regular group and PCT group. For the regular group, we applied antibiotic treatment and determined the course of treatment based on clinical symptoms, laboratory test results, medical imaging results, and bacterial culture. For the PCT group, in addition to the above treatments, antibiotic treatment was applied when the PCT level was not lower than 0.25 ng/ml and stopped when the PCT level was lower than 0.25 ng/ml. The distribution of bacterial infection sites, type of bacteria, type of antibiotics, average period of hospitalization, and course of antibiotic treatment were compared between the two groups.

RESULTSThere were no significant differences in the distribution of bacterial infection sites, type of bacteria, type of antibiotics, and average period of hospitalization between the two groups (P > 0.05). The course of antibiotic treatment for the PCT group was significantly shorter than that for the regular group (25.37 ± 11.66 vs 20.58 ± 7.53 d, P < 0.05).

CONCLUSIONUnder similar conditions of bacterial infection, antibiotic treatment of TCE-induced dermatitis based on the serum PCT level can significantly shorten the course of treatment and avoid the abuse of antibiotics.

Anti-Bacterial Agents ; therapeutic use ; Bacteria ; Bacterial Infections ; Calcitonin ; analysis ; Calcitonin Gene-Related Peptide ; Drug Eruptions ; drug therapy ; Hospitalization ; Humans ; Monitoring, Physiologic ; Protein Precursors ; analysis ; Trichloroethylene ; toxicity

AIM:To understand the clinic effect of intravitreal anti-vascular endothelial growth factor(VEGF) drugs injection in the treatment of fundus disease in the real-world study (RWS).METHODS:The clinical cases treated with anti-VEGF drugs in our department from September 2012 to June 2015 were enrolled in this study.Retrospective investigation was reviewed to the kinds of diseases, frequency, usage, efficacy, adverse reaction, and the effects on visual acuity, fundus and macular thickness which were treated with intravitreal anti-VEGF drugs injection.RESULTS:In 305 patients (340 eyes) treated with anti-VEGF drugs, 53 patients (60 eyes, 17.6%) were wet age-related macular degeneration (AMD), polypoidal choroidal vasculopathy (PCV) 16 cases (18 eyes, 5.3%), diabetic macular edema (DME) 120 cases (134 eyes, 39.4%), branch retinal vein occlusion (BRVO) secondary macular edema 61 cases (68 eyes, 20.0%), central retinal vein occlusion (CRVO) secondary macular edema 29 cases (32 eyes, 9.4%), idiopathic choroidal neovascularization (ICNV) 16 cases (18 eyes, 5.3%), high myopia with choroid neovascularization 4 cases (4 eyes, 1.2%), neovascular glaucoma 4 cases (4 eyes, 1.2%), retinal angiomatous proliferation (RAP) 1 cases (1 eyes, 0.2%) and optic papillary neovascularization 1 cases (1 eyes, 0.2%).The minimum age was 16 years old, and the maximum age 90 years old.There were 247 cases (275 eyes, 80.9%) were treated with intravitreal ranibizumab injection, 58 cases (65 eyes, 19.1%) intravitreal conbercept injection.The time number of all patients accepted anti-VEGF drugs treatment was 465, with an average of 1.7 times per eye.Which, the 3 + PRN treatment method in 98 patients (109 eyes, 32.1%), 1 + PRN treatment in 207 patients (231 eyes, 67.9%).69 cases (77 eyes, 22.6%) were used alone to receive anti-VEGF drugs therapy, 10 cases (11 eyes, 3.2%) combined with intravitreal triamcinolone injection(TA), 35 cases (39 eyes, 11.5%) combined with vitrectomy, 26 cases (29 eyes, 8.5%) combined with photodynamic treatment (PDT), 165 cases (184 eyes, 54.1%) combined with simple laser treatment.After anti-VEGF drug treatment, majority of patients' the best corrected visual acuity (BCVA), fundus and central macular thickness(CMT) were significantly improved, compared with the pre-treatment, the difference is significant (P<0.05).So that anti-VEGF drugs can effectively improve visual function and ocular fundus for fundus diseses.There were no serious adverse reactions except 3 patients appearling skin redness, itching, rash, 1 patient low low-grade fever and 1 patient acute cerebral infarction during the treatment.CONCLUSION:Intravitreal anti-VEGF drugs injection can significantly improve the visual function and ocular fundus for patients with fundus diseases, but there are still some adverse events, which should be attached great importance to medical workers.

OBJECTIVEEstablish the model of mouse with chronic hepatitis B virus (HBV) and nonalcoholic fatty liver disease (NAFLD).

METHODSTake 100 HBV transgenic, BALB/c mice of 4 weeks old, with each gender half. Then pick out 70 mice in one group to feed high-fat feed and the rest to feed normal feed. At the end of week 16, random kill 10 mice of high-fat, then liver tissue and serological detection target identification model is established in this paper. After that, divide the mice into model group and comparison group with 30 mice in each group. Feed model group with high-fat feed, comparison group with normal feed and normal group with normal feed till week 72 (including previous 16 weeks). Kill 10 mice of each group at the end of week 24, 48 and 72 respectively, fully automatic biochemical instrument detection of serum ALT, AST, TC, TG, FBG, fluorescence quantitative PCR method to detect HBV-DNA, chemiluminescence detection of HBsAg, liver biopsy after HE staining to evaluate histology change, observe mice model of dynamic evolution.

RESULTS(1) Feed high fat feed after 16 weeks, mice's weight, serum ALT, AST, TC, TG, FBG and blood biochemical indicators increased, HBV-DNA positive, liver HE staining obviously big blister fatty degeneration of liver cells and within the lobule lymphocytes infiltration, NAFLD activity score (NAS) getting close to NASH, the model of chronic HBV carries with NAFLD mouse built successfully. (2) The TC and TG values of model group in each period were higher than that of comparison group and normal group. (3) In week 24 and 72, HBV-DNA values of each group are obvious different from the other two groups and the difference can be applied to statistical significance (P < 0.05). (4) In week 48 and 72, NAS of each group are obvious different from the other two groups and the difference can be applied to statistical significance (P < 0.05).

CONCLUSIONS(1) Chronic HBV carries with NAFLD mice model can be established by HBV transgenic mice fed by high fat feed. (2) NAFLD accelerates the liver disease of the mice carrying HBV to some extent.

Animals ; Disease Models, Animal ; Fatty Liver ; complications ; pathology ; virology ; Female ; Hepatitis B virus ; genetics ; isolation & purification ; physiology ; Hepatitis B, Chronic ; complications ; pathology ; virology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Transgenic ; Non-alcoholic Fatty Liver Disease

OBJECTIVETo observe the functional changes of dendritic cells (DCs) infected in vitro by 3 recombinant adenoviruses encoding Her2/neu extracellular first-receptor domain (Her2-ECDs), full-length extracellular domain (Her2-ECD), and extracellular and transmembrane domain (Her2-TM) proteins (rAdHer2-ECDs, rAdHer2-ECD and rAdHer2-TM, respectively).

METHODSThe expressions of the target proteins were detected with Western blotting. The level of both interleukin (IL)-12 in the supernatant of in vitro cultured DCs infected with recombined adenoviruses and interferon gamma (IFN-gamma) in the supernatant of the lymphocyte populations co-cultured with DCs were determined by enzyme-linked immunosorbent assay (ELISA). The capacity of the DCs to stimulate allogeneic T lymphocyte proliferation was assessed by mixed lymphocyte reaction, and the activity of cellular toxic T lymphocytes (CTL) were investigated by MTT assay.

RESULTSHer2-ECDs, ECD and TM proteins were detected in the transfected DCs. Compared with the untransfected DCs, more abundant IL-12 production was detected in the supernatant of the DCs 5 days after transfection, but the IL-12 level showed no significant difference between the DCs infected with the 3 recombinant adenoviruses. IFN-gamma production increased gradually with passage of the time following DC-stimulated lymphocyte proliferation irrespective of infection of the DCs, and only the DCs infected with rAdHer2-TM seemed to result in significant difference in DC-mediated allogeneic T lymphocyte proliferation. The killing of breast cancer cell line with Her2 overexpression was more efficient with infected DCs priming autologous T lymphocyte to generate CTL than with uninfected DCs and those modified by SK-OV-3 cell fragment. CTL activity induced by rAdHer2-TM-infected DCs was the strongest, and breast cancer cell-killing activity was more efficient against cell line with Her2/neu-overexpression.

CONCLUSIONThe DCs infected with the recombinant adenovirus encoding Her2/neu extracellular and transmembrane domains show enhanced anti-tumor effect and induce Her2/neu-specific CTL activity.

Adenoviridae ; genetics ; Blotting, Western ; Breast Neoplasms ; genetics ; immunology ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; immunology ; Coculture Techniques ; Cytotoxicity, Immunologic ; immunology ; Dendritic Cells ; cytology ; immunology ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Extracellular Matrix Proteins ; genetics ; metabolism ; Female ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; metabolism ; Lymphocyte Culture Test, Mixed ; Lymphocytes ; cytology ; immunology ; Membrane Proteins ; genetics ; metabolism ; Ovarian Neoplasms ; genetics ; immunology ; pathology ; Receptor, ErbB-2 ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; T-Lymphocytes, Cytotoxic ; cytology ; immunology ; Transfection

Objective To study the analgesic and anti-inflammatory effects in vivo of the total alkaloids in Stephania kwangsiensis (TARS).Methods Mice were respectively ig with high,middle,and low doses of TARS.The analgesic effects of TARS were evaluated by the formaldehyde induced pain method and tail-immersion response induced by hot water in mice,and anti-inflammatory effects were observed by cotton ball granuloma,egg white induced paw swelling method in mice.Results TARS could significantly inhibit formaldehyde induced pain in mice,reduce the hind licking times,extend tail withdrawal time induced by hot water,obviously reduced the formation of granulomatous in mice;Inhibited egg white induced paw swelling.Conclusion TARS has significant analgesic and anti-inflammatory effect.