OBJECTIVETo investigate the role of transient receptor potential melastatin 8 (TRPM8) channels in migraine mechanism in rats by measuring the changes in expression of TRPM8 in the trigeminal nerve of rats with migraine.

METHODSTwenty male Sprague-Dawley rats were randomly and equally divided into a blank control group and a model group. Nitroglycerin (10 mg/kg) was injected subcutaneously in the back of the neck once a week for 5 weeks, to prepared a rat model of migraine without aura. Normal saline was injected subcutaneously instead of nitroglycerin in the control group. At 4 hours after the final injection, behavior scoring of all rats was performed, and then the trigeminal nerve ganglions of rats in both groups were collected for measurement of expression of N-methyl-D-aspartate receptor (NMDAR), protein kinase A (PKA), and TRPM8 using immunohistochemical staining, immunofluorescence, and Western blot, respectively.

RESULTSThe behavior score in each week during the rat model preparing was significantly higher in the model group than in the control group (P<0.05). The expression of NMDAR, PKA, and TRPM8 in the model group was significantly higher than in the control group (P<0.01). Both the behavior score and the expression of NMDAR were positively correlated with the expression of TRPM8 (r=0.822 and 0.794 respectively; P<0.01).

CONCLUSIONSTRPM8 may be involved in migraine mechanism probably by activation of the NMDAR pathway.

Animals ; Cyclic AMP-Dependent Protein Kinases ; analysis ; Male ; Migraine Disorders ; etiology ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; analysis ; physiology ; TRPM Cation Channels ; physiology

Aim Study the interaction between metho-trexate and levetiracetam after oral administration and hepatorenal toxicity in rats, to provide theoretical basis for the clinical rational use of two drugs. Methods Male SD rats ( 250 ± 20 ) g were selected as experi-mental animals. Rats were randomly divided into 3 groups ( 1 mg·kg-1methotrexate group, 180 mg· kg-1 levetiracetam group, combined group), 5 rats in each group. The following experiment was carried out:(1) pharmacokinetic parameters study; (2) renal ex-cretion study; (3) liver and kidney toxicity study. All samples were determined by ultra performance liquid chromatography-tandem mass spectrometry ( UPLC/MS/MS) . Results After the combination of two drugs, the Cmax, AUC0-t and Tmax of methotrexate in-creased, T1/2, Cl and MRT of methotrexate decreased, the differences was statistically significant. The Tmax, T1/2, Cl, MRT of levetiracetam had a certain upward trend, AUC0-t had a certain downward trend, the differences has no statistically significant. After combi-nation of two drugs, the renal excretion of methotrexate increased significantly, the renal excretion of levetirac-etam decreased significantly and hepatorenal toxicity increased significantly. Conclusion After combina-tion of two drugs by oral administration, Levetiracetam significantly increased methotrexate absorption in rats. Levetiracetam increased the plasma concentration and renal excretion of methotrexate and caused hepatorenal toxicity significantly. It is suggested that dose adjust-ment is necessary for combination of two drugs.

OBJECTIVETo analysis the clinic and genotype in two Chinese patients with Dyskeratosis congenita (DC).

METHODSThe two patients were characterized by mucocutaneous abnormalities (abnormal nails, lacey reticular pigmentation, and oral leukoplakia), bone marrow failure. They were diagnosed with DC. DC genes were amplified by polymerase chain reaction (PCR), including DKC1, TERT, TERC, TINF2, NOP10, NHP2, then DNA sequencing was performed for abnormal exons.

RESULTSAn abnormal peak was found in exon 6 of TINF2 gene of the two patients. DNA sequencing showed a 845G→A transition in TINF2 gene in the two patients.

CONCLUSIONWe should think about DC if the young patients with mucocutaneous abnormalities and marrow failure. TINF2 c.845G→A(R282H) does exist in the two patients. It is reported in China for the first time.

Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Dyskeratosis Congenita ; diagnosis ; genetics ; Exons ; Female ; Humans ; Infant ; Male ; Telomere-Binding Proteins ; genetics

This article describes the preparation of the N-tert-butyl-alpha-phenylnitrone (PBN) liposomes and their related characteristics. The PBN liposomes were prepared by film dispersion-supersonic method and the formula of liposomes was optimized by orthogonal uniform design. RP-HPLC was used to qualify the amount of PBN that entered into the hepatoma cells. Necrosis rate was also investigated by fluorescence activated cell sorter (FACS) after PBN liposomes transfection. Result showed that the mean particle size, entrapment efficiency, and polydispersity of the resulting PBN-liposome were 137.5 nm, 71.52% and 0.286, respectively. PBN liposomes can enter into the tumor cell stably and they have higher affinity to hepatoma cell compared with free PBN resulting in a higher necrosis rate after transfection. These results provide a potential method for early diagnosis and treatment of cancer using specific spin trapping probe targeting tumor cells.
Carcinoma, Hepatocellular ; pathology ; Cyclic N-Oxides ; chemistry ; Electron Spin Resonance Spectroscopy ; Humans ; Nanoparticles ; chemistry ; Spin Labels ; Spin Trapping ; methods ; Tumor Cells, Cultured

Objective To study the clinical value of intraoperative epicardial echocardiography ( IEE) in assessing graft-left anterior descending artery ( LAD ) of off-pump coronary artery bypass grafting ( OPCABG) . Methods IEE was used to detecte graft vessels anastomosis in 53 patients with OPCABG-LAD . Two-dimensional grey ultrasound and color Doppler ultrasound was used to show whether there was abnormal echo in proximal and distal lumen , measuring diameter and rate of stenosis . Pulse Doppler ultrasound was used to observe the blood flow spectrum . Intraoperative transient blood flow meter (TTFM) was employed to measure the pulsatility index and flow volume . Results Among the 53 patients with OPCABG-LAD ,38 cases were left internal mammary artery graft (LIMA) ,15 cases were saphenous vein graft (GSV) . Pulsatility index and flow volume showed normal by TTFM . The detection rates of graft vessels-LAD anastomosis in proximal and distal segment were 100％ using IEE , including 10 cases anastomotic plaques and 10 cases proximal plaques . Comparison of blood flow parameter of graft by IEE and TTFM in operation ,there was no significant difference in LIMA grafts ( P =0 .091) ,but the correlation was statistically significant ( r = 0 .809 , P < 0 .001 ) ;the difference in GSV grafts had no statistical significance ( P = 0 .821 ) ,but the correlation was statistically significant ( r = 0 .684 , P = 0 .005 ) . Conclusions IEE clearly displays the lumen of graft vessel and LAD ,and measures the hemodynamic indexes . It provides an intuitive ,accurate and convenient method for detecting the patency of the graft vessels during OPCABG .

8-hydroxy-2 deoxyguanine (8-OHdG) as a biomarker of DNA oxidative damage,its expression level is closely related to the mutation of many oncogenes,and plays an important role in the occurrence,development and prognosis of cancer.8-OHdG can be an effective biomarker for the early screening,efficacy monitoring and prognosis evaluation of oxidative injury disease,and play an important role in clinical practice.This paper summarizes the previous clinical studies of 8-OHdG,which is intended to provide implications for its clinical application.

Objective To develop a method for the detection of 8-hydroxy deoxyguanosine (8-OHdG) in urine and to explore its value in tumor-assisted diagnosis.Methods The chromatographic separation was performed on Agilent HC-C1sColumn (4.6 mm ×250 mm,5 μm)with the mobile phase of 90％ 50 mmol · L-1 NaH2PO4aqueous solution and 10％ methanol at a flow rate of 1.0 mL · rain-1 The detection wavelength was 300 nm.The injection volume was 50 μL The concentration of 8-OHdG in the urine of patients with cancer was detected by HPLC.The control group was healthy person and the patients were divided into two groups:before and after the operation,before and after chem-otherapy.Results The method had a good linear relationship.The limit of detection was 0.1 μg · mL-1.The RSDs of imer-day and intra-day precision were less than 2％.The spiked recovery rates were more than 90％.The level of 8-OHdG in lung cancer group was (5.49 ± 5.48)μg · mL-1,the level of 8-OHdG in lung cancer group was (0.51 ± 0.63) μg · mL-1,the level of 8-OHdG in healthy group was (1.16 ± 1.65) μg · mL-1.The concentration of 8-OHdG in preopretive group was significantly higher than that in postoperative group and control group (all P ＜ 0.05).Conclusion 8-OHdG detected by HPLC has high accuracy and specificity in tumor diagnosis and evaluation of chemotherapy efficacy.

A case series report was conducted to analyze the cases of children with multiple sclerosis(MS)who received fingolimod for the first time.Four Chinese children with MS were reported.A 9-year-old girl weighted 50 kg with low basal heart rate developed asymptomatic bradycardia.The other 3 children did not experience cardiac adverse reactions.Cardiac safety of the first dose of fingolimod in children with MS was also evaluated.We suggest that special attention should be paid to patients with slow basal heart rate,young age and low body weight.In order to promote its safe use at initiation,the monitoring time can be extended appropriately and night monitoring can be strengthened.

Background: Right upper lobectomy (RUL) for lung cancer with different dissecting orders involves the most vari-able anatomical structures, but no studies have analyzed its effects on postoperative recovery. This study compared the conventional surgical approach, VAB (dissecting pulmonary vessels first, followed by the bronchus), and the alter-native surgical approach, aBVA (dissecting the posterior ascending arterial branch first, followed by the bronchus and vessels) on improving surgical feasibility and postoperative recovery for lung cancer patients. Methods: According to the surgical approach, consecutive lung cancer patients undergoing RUL were grouped into aBVA and VAB cohorts. Their clinical, pathologic, and perioperative characteristics were collected to compare periop-erative outcomes. Results: Three hundred one patients were selected (109 in the aBVA cohort and 192 in the VAB cohort). The mean operation time was shorter in the aBVA cohort than in the VAB cohort (164 vs. 221 min, P < 0.001), and less blood loss occurred in the aBVA cohort (92 vs. 141 mL, P < 0.001). The rate of conversion to thoracotomy was lower in the aBVA cohort than in the VAB cohort (0% vs. 11.5%, P < 0.001). The mean duration of postoperative chest drainage was shorter in the aBVA cohort than in the VAB cohort (3.6 vs. 4.5 days, P = 0.001). The rates of postoperative complica-tions were comparable (P = 0.629). The median overall survival was not arrived in both cohorts (P > 0.05). The median disease-free survival was comparable for all patients in the two cohorts (not arrived vs. 41.97 months) and for patients with disease recurrences (13.25 vs. 9.44 months) (both P > 0.05). The recurrence models in two cohorts were also comparable for patients with local recurrences (6.4% vs. 7.8%), distant metastases (10.1% vs. 8.3%), and both (1.8% vs. 1.6%) (all P > 0.05). Conclusions: Dissecting the right upper bronchus before turning over the lobe repeatedly and dissecting veins via the aBVA approach during RUL would promote surgical feasibility and achieve comparable postoperative recovery for lung cancer patients.

OBJECTIVETo investigate the molecular mechanism by which LKB1 regulates epithelial-mesenchymal transition (EMT) in Peutz-Jeghers hamartoma and intestinal epithelial cells.

METHODSImmunohistochemistry was used to detect gene expression of LKB1, E-cadherin, and vimentin in 20 hamartoma tissues and 10 normal intestinal tissues, and collagen fiber deposition was analyzed using Masson trichrome staining. Normal intestinal epithelial NCM460 cells were transfected with LKB1 shRNA plasmid or negative control via lentiviral vectors, and the role of LKB1 in cell polarization and migration were determined using CCK8 and Transwell assays. Western blotting, quantitative real-time PCR (qPCR) and immunofluorescence were used to assess the alterations of EMT markers in the cells with LKB1 knockdown.

RESULTSCompared with normal intestinal tissues, hamartoma polyps showed significantly decreased LKB1 and E-cadherin expressions and increased vimentin expression with increased collagen fiber deposition. The cells with LKB1 knockdown exhibited enhanced cell proliferation and migration activities (P<0.01). Western blot analysis, qPCR and immunofluorescence all detected decreased E-cadherin and increased N-cadherin, vimentin, Snail, and Slug expressions in the cells with LKB1 knockdown.

CONCLUSIONs LKB1 deficiency triggers EMT in intestinal epithelial cells and Peutz-Jeghers hamartoma, suggesting that EMT can serve as the therapeutic target for treatment of Peutz-Jeghers syndrome.