Aim To investigate the effect of oxymatrine on lipid metabolism regulated genes in liver in fat-in-duced insulin resistance in ApoE -/- mice.Methods Seventeen C57BL/6J male mice were selected in normal control group.Sixty-eight ApoE -/- mice with high fat diet for 1 6 weeks,were randomly divided into model group,oxymatrine low,middle and high dose groups.Then they were gavaged for 8 weeks.Body weight and general biochemical indicators were deter-mined in mice.The mRNA and protein expression lev-els of LPL,FAT/CD36,CPT1 ,UCP2,SREBP-1 c,FAS and ACC were examined by real-time PCR and West-ern blot in the liver.Results Compared with model group,oxymatrine reduced body weight(BW),fasting
blood glucose (FBG),cholesterol (TC ),triglyceride (TG),free fatty acids(FFA),fasting plasma insulin (FINS)and insulin resistance index(HOMA-IR)(P <0.05),while improved glucose infusion rate (GIR). Oxymatrine down-regulated the mRNA and protein ex-pression of LPL,FAT/CD36,UCP2,SREBP-1 c,FAS and ACC(P <0.05),and up-regulated the mRNA and protein expression of CPT1 in varying degrees (P <0.05).Conclusion Oxymatrine can regulate the ex-pression of lipid metabolism regulated genes in liver and improve insulin resistance in ApoE -/- mice in-duced by high fat diet.

Objective To investigate the relationship between prothrombin fragment 1+2 (F1+2) and peripherally inserted central catheter (PICC) associated thrombosis in cervical cancer patients, and provide certain clinical basis of early prevention in peripherally inserted central catheter associated thrombosis in cervical cancer patients. Methods One hundred and forty cervical patients with PICC were enrolled in this study, and they were divided into thrombosis group (35 patients) and non-thrombosis group (105 patients). The level of F1+2 was examined using enzyme-linked immunoassay, and was analyzed according to the clinic features. Results The level of F1+2 was correlated with clinical stage (r = 0.640, P = 0.004);but was not correlated with age, type of tumor and concurrent radiochemotherapy (P>0.05). The level of F1+2 in thrombosis group was (520.343 ± 121.759) pmol/L, in non- thrombosis group was (388.361 ± 104.873) pmol/L, and there was significant difference (P =0.001). The multi-factors Logistic analysis showed that the level of F1+2 (OR=1.011, P=0.001) and age (OR = 21.025, P = 0.031) were independent risk factors for the PICC associated with thrombosis in cervical cancer. Conclusions The level of F1+2 is closely related with clinical stage and PICC associated thrombosis, and it is an independent risk factor for the PICC associated with thrombosis in cervical cancer.

Lymphangiomas are malformations of the lymphatic system, which is a common disease in children, usually superficial in body, but seldom seen in visceral organs. We present the case of a child with lymphangioma of the gallbladder. Also we reviewed the literatures. A 2-year-6-month-old boy complained of progressing difficulty in walking for 6 months. Cerebral MRI showed abnormal signals in the white matter suggesting leukodystrophy. Ultrasound of abdomen showed a mixed-echoic mass in the site of gallbladder. CT scan showed an enlarged gallbladder with increased density. Bloodcounting, liver function, and alpha fetal protein were within normal range. Exploratory laparotomy was done in order to rule out malignancy. The liver was found normal at surgery, and the gallbladder looked enlarged and deformed. The gallbladder wall was thick and edematous, and adherent with liver. The gallbladder was excised. Pathological examination gave the diagnosis as lymphangioma of the gallbladder wall. The postoperative recovery was uneventful. There was no evidence of recurrence. Searching in literatures, three cases of lymphangioma of gallbladder in adults were found. According to the pathology of lymphangioma and the anatomy of gallbladder, the outcome of this disease should be benign and may be symptomless throughout the life. No death was reported due to lymphangioma of gall bladder, nor severe complications endangering life at any age. The only indication for surgery in the reported cases was to rule out the risk of being malignancy. If MRI or other means can make a definite preoperative diagnosis of lymphangioma, being a stable lesion, surgery would be unnecessary unless there is a progressive organic obstruction causing repeated infection or persistent abdominal pain.
Child, Preschool ; Gallbladder Neoplasms ; diagnosis ; Humans ; Lymphangioma ; diagnosis ; Male

Objective: The current study was designed to observe and discuss the variation of neuroblastoma cells in morphology, cell cycle and telomerase gene (hTR) expression induced by retinoids. Methods: Two newly-synthesized retinoids, R9158 and R8605, were supplemented into the media of neuroblastoma cells, SH-SY-5Y and SK-N-SH, with the concentration of 10- 6 mol/L as the treatment group. Meanwhile the same supplements of DMSO were as the control group. Pre-and post-supplement cells were observed for morphological changes, cell cycle was analyzed by flow cytometry (FCM) and hTR expression was analyzed by in situ hybridization chemistry (ISHH). Results: 1) The shape of SH-SY-5Y cells changed significantly at the 5th day of R9158 and R8605 treatment, having multiple neurite extensions just as mature neuron. However no shape change of SK-N-SH cells was found in observation for 7 days. 2)For SH-SY-5Y cells, the percentage of Stage G1, S, G2/M differed much between the treatment group and the control group at 5th day when the cell cycle of the treatment group was retarded at Stage G1 and the ratio of Stage decreased significantly. For SK-N-SH cells, however, there was little variation of cell cycle status in the treatment group. 3)The SH-SY-5Y cell numbers of hTR positive expression in the treatment groups decreased significantly at the 5th day. No decrease was found in the R9158 and R8605 groups of SK-N-SH cells in the treatment groups. Conclusions: During the differentiation of RA-sensitive cells induced by retinoids, the telomerase expression was depressed and the cell cycle retarded;whereas the RA-resistant cells changed little in morphology, telomerase expression and cell cycle as well. The telomerase gene associated with other genes would be an important regulated target in the differentiation induced by retinoids.

OBJECTIVETo investigate the efficiency of different angle projection radiograph in diagnosing multiple canals of mandibular first premolars.

METHODSEighty-eight mandibular first premolars that needed endodontic treatment in vivo were selected. The radiograph was taken at a horizontal angles of 0, 20-30 degrees from mesial or distal direction of the tooth preoperative. According to the different angle projection radiographic evaluation, radiographic diagnosis of multiple canals were suspected. After the root canal orifice were detected and located with K files, the root canals were prepared using crown-down technique and obturated using cold lateral condensation technique. The configurations and numbers of root canals were identified and recorded. The postoperative radiograph was taken. According to configurations and postoperative radiograph, the root canal configuration was classified into type I -V.

RESULTSIn eighty-eight mandibular first premolars, 31 multiple canals teeth were suspected in preoperative radiographic diagnosis, 30 multiple canal teeth were detected under clinical evaluation. In case of 31 multiple canal teeth suspected by preoperative radiographic diagnosis, 3 teeth were not found multiple canals under clinical evaluation. While in case of 2 teeth of one canal suspected by preoperative radiographic diagnosis, multiple canal was detected under clinical evaluation. In 31 multiple canal teeth suspected by preoperative radiographic diagnosis, 13 teeth were detected at a horizontal angles of 0 degree and 25 teeth were detected at a horizontal angles of 20-30 degree, 7 teeth were suspected at different angle projection. Clinical detected rate of multiple canal in mandibular first premolar were 34.09% (30/88), 70.00% of which were type IV and V.

CONCLUSIONDifferent angulation radiograph of preoperative will assist to increase clinical detection rate of multiple canals in mandibular first premolar.

Bicuspid ; Dental Pulp Cavity ; Humans ; Incisor ; Mandible ; Maxilla ; Molar ; Postoperative Period ; Root Canal Therapy

OBJECTIVETo establish the BGC-823/WTX-EGFP gastric cancer cell line with stable expression of Wilms tumor gene on the X chromosome (MTX) for functional analysis of WTX gene.

METHODSThe full-length WTX cDNA was amplified from human embryonic kidney 293FT cells and cloned into the pEGFP-N1 vector containing the reporter gene of green fluorescence protein. The recombinant pEGFP-WTX expression vector, after digestion by restriction enzyme to identify the size of target gene fragment, was transfected into 293FT cells and the expression of fluorescent reporter gene was observed under fluorescence microscope. pEGFP-WTX vector was transfected into human gastric cancer BGC-823 cell line to establish BGC-823/WTX-EGFP cell line stably expressing WTX. Quantitative RT-PCR and immunocytochemical staining were used to detect the expression of WTX in both BGC-823/WTX-EGFP and control BGC-823 cells.

RESULTSThe recombinant pEGFP-WTX plasmid was successfully constructed and verified by PCR and sequencing. The mRNA and protein expressions of MTX were significantly increased in BGC-823/WTX-EGFP cells as compared with those in the control cells.

CONCLUSIONThe full-length WTX expression vector (pEGFP-WTX) and BGC-823/WTX-EGFP gastric cancer cell line have been successfully established to facilitate further functional study of WTX gene.

Cell Line ; Cell Line, Tumor ; Chromosomes, Human, X ; genetics ; DNA Restriction Enzymes ; DNA, Complementary ; Genes, Wilms Tumor ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Plasmids ; Stomach Neoplasms ; genetics ; Transfection

Objective To compare the anesthetic effect and safety of remimazolam and propofol on patients underwent video-assisted thoracoscopic surgery for lung cancer.Methods Clinical data of patients with lung cancer underwent video-assisted thoracoscopic surgery were retrospectively collected.Remimazolam group was anesthetized by remimazolam,and propofol group was anesthetized by propofol.The changes in mean arterial pressure(MAP)and heart rate(HR)were compared between the two groups of patients before anesthesia induction(T0),after 5 min of tracheal intubation(T1),after 1 h of surgery(T2),during thorax closure(T3)and at 5 min after extubation(T4).The sedation onset time,recovery time and extubation time in the two groups were recorded.Stress response indicators[adrenocorticotropic hormone(ACTH),cortisol(Cor)]were compared at T0 and T4.Ramsay sedation score(RSS)was used to assess the sedation degree at T4.Visual analogue score(VAS)was applied to evaluate the pain degree at 2,12 and 24 h after surgery,and the perioperative anaesthesia-related adverse events were observed.Results There were 58 cases in remimazolam group and 64 cases in propofol group.The MAP values at T1 in remimazolam group and propofol group were(85.03±4.37)and(78.24±4.48)mmHg;at T2 were(80.39±3.95)and(75.49±4.11)mmHg;at T3 were(84.43±4.02)and(79.59±3.97)mmHg;the HR values at T2 were(76.44±5.75)and(72.39±6.03)beat·min-1,the difference were all significant(all P＜0.05).The sedation onset times in remimazolam group and propofol group were(62.45±6.27)and(72.33±7.19)s;the recovery times were(7.22±1.23)and(8.24±1.48)min;the extubation times were(8.34±1.50)and(10.09±1.83)min;the RSS scores at T4 were(2.03±0.39)and(1.88±0.35)points,the difference were all significant(all P＜0.05).The total incidence rates of anesthesia-related adverse events in remimazolam group and propofol group were 6.90％and 21.88％,respectively(P＜0.05).Conclusion Both remimazolam and propofol can play a good sedative effect during lung cancer video-assisted thoracoscopic surgery anesthesia.Remimazolam anesthesia has more stable intraoperative hemodynamics,faster onset and elimination,and higher safety.

OBJECTIVETo compare different methods commonly used for titering adenovirus and analyze the advantages and limitations of each method.

METHODSFour recombined adenoviruses (Ad-G-AT2R-EGFP, Ad-CMV-EGFP, Ad-mif-shRNA-EGFP and Ad-CBA-GFP) were amplified and purified, and each was titered by optical absorbance, real-time PCR, green fluorescent protein (GFP)-labeled method, immunoassay, and cytopathic effect (CPE). The results were then comparatively analyzed.

RESULTSNo significant difference was found in the titer amounts derived from GFP-labeled method, immunoassay, and cytopathic effect method (P>0.1). A positive correlation was noted in the titer amounts determined by real-time PCR and immunoassay (r=0.965), even though the value (vg/ml) obtained by real-time PCR was 10 times higher than that by immunoassay (ifu/ml).

CONCLUSIONGFP-labeled method and immunoassay allow rapid determination of the adenoviral titer. Real-time PCR can not directly determine the real infectious titer of the adenovirus, but the result is well correlated to that of immunoassay and reflects, though indirectly, the actual infectious titer of adenovirus. Considering the procedural convenience and shorter time consumption, real-time PCR is still a practical method for adenoviral titration.

Adenoviridae ; isolation & purification ; metabolism ; physiology ; Capsid Proteins ; metabolism ; DNA, Viral ; isolation & purification ; Green Fluorescent Proteins ; metabolism ; Real-Time Polymerase Chain Reaction ; methods ; Viral Plaque Assay ; methods ; Virus Replication

OBJECTIVETo investigate the expression characteristics of Gluc-Fluc dual luciferase plasmid after its transfection into MB49 bladder cells.

METHODSpAAV2neoCAG-Gluc-2A-Fluc and pAAV2neo-Gluc plasmids were separately transfected into MB49 cells via LipofectamineTM2000. The Gluc activity in the cell culture supernatant and the Fluc activity in the cells were detected by luminometer and Lumina Imaging system.

RESULTSThe luminometer result showed that the activity of Gluc in the supernatant increased gradually in a cell number- and time-dependent manner, while Fluc activity in the cells increased with the cell number but not with time. The Lumina Imaging system showed that Gluc-Fluc was successfully expressed in MB49 bladder cells and cell lines with stable Gluc-Fluc expression were obtained after G418 selection.

CONCLUSIONGluc in the dual luciferase plasmid retains its expression characteristics. Due to the advantages of Fluc in localization in living imaging and the easy quantitative detection of Gluc, the dual luciferase plasmid, after transfection in MB49 bladder cells, allows reliable and dynamic detection of tumor growth in animal models.

Animals ; Cell Line, Tumor ; Genetic Vectors ; Luciferases ; genetics ; metabolism ; Mice ; Plasmids ; genetics ; metabolism ; Transfection

To investigate the distribution of ticks and to detect the new bunyavirus in ticks and infection rate in animals in Shanghai,outdoor free ticks were captured by flag method and ticks on animals were collected by animal capture.SFTSV in ticks were detected by real-time RT-PCR.Total antibodies against SFTSV in host animal sera were tested by double antigen sandwich method.Results showed that during 2012-2014,free ticks were captured in Chenshan Park and Jinshan Island,which were all Haemaphysalis longicornis.Ten of seventeen districts found ticks on animals which were dominantly Rhipicephalus sanguineus.No nucleic acid of SFTSV was detected in 143 ticks.Main host animal for ticks was dog,sheep was in the second.Sera in 198 dogs from 6 urban districts,120 swine from Pudong and Fenxian districts and 36 sheep from Chongming District were all SFTSV antibody negative.Ticks were not found with SFTSV in Shanghai during 2012-2014.No SFTSV infection was found in host animals.Therefore,there is no evidence that Shanghai is the natural foci of SFTSV.Further surveillance and investigations should be carried out in the future.