Objective In order to understand the differentially expressed genes and explore the effects on mechanism of gene expression induced by arsenic trioxide. Methods The mRNA was isolated from human HepG2 cells treated with arsenic trioxide( 5?mol/L ) and DMSO, respectively, then cDNA was synthesized. After restriction enzyme Rsa Ⅰ digestion, small sizes cDNA were obtained. Then tester cDNA was subdivided into two portions and each was ligated with different cDNA adaptor. After tester cDNA was hybridized with driver cDNA twice and underwent nested polymerase chain reaction (PCR) twice, the DNA fragment was subcloned into T/A plasmid vectors to set up the subtractive cDNA library. Amplification of the library was carried out with E. coli strain JM109. The cDNA was sequenced and analyzed in GenBank with Blast search after colony PCR. Results The forward subtracted cDNA library from HepG2 cell line induced by arsenic trioxide was successfully constructed. The sequencing analysis showed that there were eight clones contained ferritin H(L) chain in the library. Conclusion Arsenic trioxide can induce the up expression of ferritin H(L) chain protein in HepG2 cells, indicated that the ferritin H(L) chain may play certain role in the mechanism of anti-arsenical cytotoxicity in liver.

Objective To study cloning and the primary function of a new gene AsTP2 transactivated by arsenic trioxide. Methods Using suppression subtractive hybridization (SSH) technique, the mRNA was isolated from HepG2 cells treated with arsenic trioxide (5?mol/L) and 0.9 percent sodium chloride, respectively, then cDNA was synthesized. SSH method was employed to analyze the differentially expressed DNA sequences between the two groups. From the subtractive cDNA library of genes transactivated by arsenic trioxide, the coding sequence of a new gene was obtained by bioinformatics method, and amplified by the method of reverse transcription polymerase chain reaction (RT-PCR). Results The novel gene was named as AsTP2, which was logged in the GenBank with the accession number AY744366. AsTP2 of 1119 nucleotides (nt), coding a protein of 372 amino acid residues (aa). Conclusion A new gene has been recognized as the new target transactivated by arsenic trioxide. The results will give a new clue to explore the molecular carcinogenic mechanism of inorganic arsenic.

Objective To compare the clinical effects of HC video laryngoscope and fiberoptic bronchoscope (FOB)in guidance of endotracheal intubation for patients undergoing cervical surgery.Meth-ods A total of 50 patients (ASA I or II)with cervical vertebra injury,nerve root cervical spondylopathy or cervical spondylotic myelopathy in the first affiliated hospital of Wenzhou medical university were selected, all of whom were undergone selective cervical operations between March 2014 and June 2015.The patients were randomly divided into two groups (n =25):HC video laryngoscope group (group H)and FOB group (group F).After induction of anesthesia,HC video laryngoscope and FOB were used for tracheal intuba-tion.⑴ Intubation time and success rates of intubation were recorded,and visual analogue scale (VAS) was used to evaluate the difficulty of intubation;⑵ Mean arterial pressure (MAP),heart rate (HR),and rate-pressure product (RPP)before incubation (Tb),immediate after intubation (T0),and 1min after in-cubation (T1)were recorded;(3)Intubation-related complications were recorded.Results ⑴ The dura-tion of intubation (19.7 ±7.1 )s in the group H was significantly shorter than that of group F (51.9 ± 19.2)s (P <0.05).The one-time success rate of intubation in group H (92%)was higher than that of group F (64%),with statistically significant differences (P <0.05).Intubation was easier in group H (P <0.05).⑵ MAP,HR and RPP of group H at T0 were higher than those of group F.And at T1,there were no statistical differences between groups in MAP,HR and RPP (P >0.05).⑶ The incidence of complications (sore throat)was lower in group H (P <0.05).Conclusions Compared to FOB,HC vid-eo laryngoscope-guided selective cervical operation has the characteristics of higher success rate of intubat-ion,shorter operation time,and easier operation,more stable hemodynamics and lower incidence of intuba-tion complications.Therefore,HC video laryngoscope is a safe and effective method in the intubation for pa-tients undergoing cervical surgery.

Objective To study the effect of MRCP for operation difficulty assessment before laparoscopic cholecystectomy. Methods 110 patients with gallbladder stone were randomly selected from March 2011 to July 2012 in our hospital,and all the patients were given LC operation after examined by MRCP. Regarded the average operation time 30 min as a criterion,when the operation time was no more than 30 min,the operation was considered as normal operation;when the operation time was more than 30 min, the operation was considered as op-eration of a certain degree of difficulty. Made a correlation analysis between LC operation difficulty and age,gender,location,volume of gall-bladder,angle of ductus cysticus, thickness of gallbladder wall, circumstances surrounding the cystic duct, length of ductus cysticus, and whether there were calculus in the cervical part of gallbladder. Results The difficulty level of LC operation is related to length of ductus cys-ticus and whether there were calculus in the cervical part of gallbladder,and there is no correlation with age,gender,location,volume of gall-bladder,angle of ductus cysticus,thickness of gallbladder wall,circumstances surrounding the cystic duct. Conclusion Routinely MRCP test in preoperative patients is of important significance to the prediction of LC operation difficulty.

Objective To investigate the relationship between metabolites of sodium arsenite and sodium dihydrogen arsenate with related metabolic enzymes in kidney of male rats.Methods According to body mass,thirty-five male Wistar rats(body mass 150-190 g) were divided into 7 groups by random number table.Control group drank deionized water; the contents of iAsⅢ in low,medium and high arsenite groups and the contents of iAsv in low,medium and high of sodium dihydrogen arsenate groups were 2.2,6.7 and 20.0 mg/kg,respectively.After 3 months,kidneys were collected and stored at-80 C; high performance liquid chromatography and hydride genesis atomic fluorescence spectroscopy (HPLC-HGAFS) was used to determine the level of arsenic metabolites in kidney,and enzyme-linked immunosorbent assay was used to detect and analyze the content or the activity of metabolic enzymes,meanwhile correlation studies between the level of metabolites and the activity of metabolic enzymes were carried out.Results The differences of total arsenic (TAs),dimethyl arsenic acid (DMA),monomethyl arsenic acid (MMA) and methyl transferase enzyme activity in kidneys of rats between groups were statistically significant (F =1874.672,H =33.513,31.002,F =79.607,all P ＜ 0.01).The TAs[(526.52 ± 25.56),(1 654.00 ± 101.55),(1 904.24 ± 104.76)μg,/kg] and DMA[(323.20 + 16.13),(1 444.40 ± 113.81),(1 765.40 ± 104.39)μg/kg] of sodium arsenite in low,medium and high dose groups were higher than those of the corresponding sodium dihydrogen arsenate groups [(235.70 ± 6.23),(471.05 ± 18.32),(1 677.40 ± 83.29)μg/kg,and(0.00 ± 0.00),(1.75 ± 0.16),(410.50 ± 19.76)μg/kg,P ＜ 0.0024 or ＜ 0.05] ; the MMA[(4.02 + 0.86),(4.20 ± 0.65),(4.04 ± 0.80)μg/kg] of sodium arsenite in low,medium and high dose groups were lower than those of the corresponding sodium dihydrogen arsenate groups[(98.90 ± 9.59),(376.50 ± 15.41),(1 131.90 ± 74.26) μg/kg,all P＜ 0.05]; the methyl transferase enzyme activities[(7.80 ± 0.93),(5.55 ± 0.49),(3.56 ± 0.26)U/g] of sodium arsenite in low,medium and high dose groups were lower than those of the corresponding sodium dihydrogen arsenate group[(11.59 ± 0.93),(8.93 ± 0.88),(6.52 ± 1.04)U/g,all P ＜ 0.0024].The DMA of sodium arsenite in low,medium and high dose groups,the MMA of sodium dihydrogen arsenate in medium and high dose groups were positively correlated with those of TAs in each group(r =0.970,0.984,0.997,0.947,0.961,all P ＜ 0.05).Conclusions Effects of sodium arsenite and sodium dihydrogen arsenate on arsenic metobdites and related metabolic enzymes in kidney of rats are different.The function of sodium dihydrogen arsenate in promoting methyl transferase activity is stronger than that of sodium arsenite,which affects the amount and distribution of arsenic methylation metabolites in kidney.

Objective To investigate the effects of repetitive ultraviolet A(UVA) radiation on the endocytosis and intracellular degradation of extracellular elastin by human dermal fibroblasts(HDFs), and to explore the mechanism responsible for elastin accumulation in photoaging skin. Methods Cultured HDFs were divided into two groups:repetitive UVA radiation group treated with UVA radiation(9.9 J/cm2)once a day for seven consecutive days to establish a chronic photodamage model, and blank control group receiving no treatment. For the verification of the model, flow cytometry was performed to detect cell cycle, and senescence-associated β-galactosidase staining to determine the degree of cell senescence. Fluorescent tracer technique and conjugated polymer-based fluorescence quenching technique were conducted to observe the endocytosis and intracellular degradation of extracellular elastin in culture media by HDFs in these groups. Results Compared with the blank control group, the repetitive UVA radiation group showed increased proportions of G1-phase cells and senescent cells, which confirmed the successful establishment of chronic photodamage model in the repetitive UVA radiation group. After coculture with elastin for 24, 48 and 72 hours, the endocytosis rate of elastin was 10.0% ± 1.4%, 27.8% ± 4.2% and 39.9% ± 4.1% respectively in the blank control group, 9.9% ± 1.6%, 28.3% ± 5.1% and 42.0% ± 5.7% respectively in the repetitive UVA radiation group, with no significant difference between the two groups at the three time points (all P > 0.05). The percentage of cells showing intracellular degradation of extracellular elastin was significantly lower in the repetitive UVA radiation group than in the blank control group (4.2% ± 1.1% vs. 7.7% ± 0.9% at 24 hours, 16.5% ± 2.4% vs. 22.8% ± 1.8% at 48 hours, 26.7% ± 2.6% vs. 33.9% ± 3.1% at 72 hours, all P < 0.05). Conclusions UVA radiation at 9.9 J/cm2 for 7 consecutive days can decrease the intracellular degradation of extracellular elastin by HDFs, but has no obvious effect on endocytosis of elastin.

OBJECTIVETo study the toxicity of Cinnabar and Cinnabar-containing traditional medicines (Zhusha Anshenwan) comparable to common mercurials.

METHODThe toxicity of methylmercury (MeHg), mercuric chloride (HgCl2), Cinnabar and Zhusha Anshenwan was studied in cultured human liver HL-7702 cells and in mice following acute and subacute exposures.

RESULTThe 50% lethal concentrations (LC50) of MeHg, HgCl2, Cinnabar and Zhusha Anshenwan in human liver HL-7702 cells were 4.4, 9.2, 2460, 4050 mg x L(-1), respectively . Oral cinnabar at a dose of 20 g x kg(-1) (clinical dosage 250 times) did not kill mouse, but no mouse could survive MeHg at a dose of 0.1 g x kg(-1) or HgCl2 at a dose of 0. 15 g x kg(-1). Subacute toxicity experiment indicated that HgCl2 retarded body weight gain with significant accumulation of Hg in the liver and kidney. In comparison, mercury accumulation after Cinnabar and Zhusha Anshenwan was insignificant. No apparent hepatic and renal dysfunctions were evident under the experimental conditions, but the metallothionein-2 mRNA levels were much higher in HgCl2 group than in other groups.

CONCLUSIONCinnabar and Zhusha Anshenwan are much less toxic than MeHg and HgCl2.

Animals ; Female ; Gene Expression ; drug effects ; Kidney ; drug effects ; physiology ; Liver ; drug effects ; physiology ; Male ; Mercuric Chloride ; administration & dosage ; adverse effects ; Mercury Compounds ; administration & dosage ; adverse effects ; Methylmercury Compounds ; administration & dosage ; adverse effects ; Mice ; Mice, Inbred BALB C ; Random Allocation

OBJECTIVETo evaluate radial artery (RA) and internal mammary artery (IMA) grafts in coronary artery bypass and the use of color Doppler ultrasound in the peri-operative evaluation of IMA and radial-ulnar collateral circulation.

METHODSFrom June 1998 to June 2000, sixty cases of coronary bypass revascularization with RA and IMA were performed. Preoperatively, the radial-ulnar collateral circulation was evaluated with the modified Allen's test, color Doppler ultrasound and noninvasive oxygen saturation measurement. The IMA lumen and blood flow were measured at the first intercostal space with color Doppler ultrasound preoperatively and postoperatively.

RESULTSOne patient (1.7%) died of serious cardiac arrhythmia on the fourth postoperative day. There were no arterial graft harvest related complications. Before harvesting, the ulnar artery blood flow was 30.78 +/- 9.71 ml/min, and it increased to 43.36 +/- 13.98 ml/min (40.87% increase, P < 0.01) after the operation. Compared with the baseline, there was no obvious change of IMA blood flow postoperatively (P > 0.05), but the systolic/diastolic flow ratio markedly decreased from 8.57 +/- 3.98 ml/min to 3.41 +/- 4.87 ml/min (P < 0.01).

CONCLUSIONSArterial grafts can be safely used for coronary bypass revascularization with good results. The ulnar artery blood flow can increase compensatively after RA harvesting. The diastolic blood flow of grafted IMA markedly increased postoperatively. Color Doppler ultrasound was very helpful both in evaluating the radial-ulnar collateral circulation before RA harvesting and in assessing the patency of the grafted IMA after coronary artery bypass grafting (CABG).

Adult ; Aged ; Coronary Artery Bypass ; methods ; Coronary Circulation ; Humans ; Mammary Arteries ; transplantation ; Middle Aged ; Radial Artery ; transplantation

Objective: To demonstrate the effects of symbiotic bacteria from lettuce on inactivation of norovirus（NV）. Methods: Symbiotic bacteria were isolated from the lettuces sampled from farmlands and supermarkets. NV mixed with symbiotic bacteria was set as the experimental group,without symbiotic bacteria as the control group. After the inactivation by high temperature,ultraviolet light（UV）and chlorine dioxide,the ratio of NV amount in the experimental group and the control group was calculated to evaluate the effects of symbiotic bacteria. The mechanism of symbiotic bacteria was revealed by detecting their effects on the protection of viral capsid protein from UV and on the adsorption of NV. Results: Eleven symbiotic bacteria were identified from lettuces,all of which were bacilli,mainly Pseudomonas. Ten symbiotic bacteria could improve the heat-resistant ability of NV,with Microbacterium oryzae,Cupriavidus taiwanensis（SC061204）,Pseudomonas furukawaii,Enterobacter tabaci and Pseudomonas resinovorans（SC061211）more significant. Eleven symbiotic bacteria could improve anti-UV ability of NV,with Pseudomonas putida,Microbacterium oryzae and Enterobacter tabaci more significant. Only one strain of Pseudomonas putida could improve anti-chlorine dioxide ability of NV（Class I hazard）. Pseudomonas putida,Microbacterium oryzae and Enterobacter tabaci could significantly reduce the damage of NV capsid protein. Nine symbiotic bacteria could promote NV adsorption into lettuces,with the promotion rates ranged from 1.04% to 46.73%;while Pseudomonas putida and Pseudomonas resinovorans（SC061211） could restrain NV absorption,with the promotion rates of -6.50% and -19.85%. Conclusion Symbiotic bacteria from lettuce may enhance the anti-inactivation of NV by protecting capsid protein and promoting adsorption of NV. It is recommended to control the presence of symbiotic bacteria in the process of inactivating NV.

Objective To analysis the distribution of posterior corneal astigmatism (PA) and evaluate the influence of keratometric astigmatism (KA) and total corneal astigmatism (TCA) on the calculation of Toric intraocular lens (Toric IOL) in patients with age-related cataract (ARC) and high corneal astigmatism.Methods An observational study design was adopted.Pentacam was used to measure 200 eyes of 181 patients with ARC and KA＞0.75 D in Affiliated Hospital of Nantong University from August 2016 to April 2017.KA,PA,TCA and anterior corneal astigmatism (AA) were recorded.The astigmatism magnitude and axis of PA was studied.The subjects were divided into astigmatism with the rule group,astigmatism against the rule group and oblique astigmatism group according to the axis of AA.The correlations of decomposition values between PA and AA or KA and TCA in each group were analyzed by Pearson linear correlation analysis.The difference of decomposition values between KA and TCA in each group was compared by paired sample t test.The type and axis of Toric IOL were calculated by online formula according to KA and TCA.This study followed the Declaration of Helsinki and written informed consent was obtained from each patient prior to any medical examination.Results The mean astigmatic magnitudes of PA was -0.32 D×93.1°.PA exceeded 0.5 D in 22 eyes (11％).The steepest posterior corneal meridian was vertically aligned in 163 eyes (81.5％).The decomposition value KP(0) and KP (45) of PA were positively correlated with those ofAA (r=0.480,P＜0.001;r=0.251,P＜0.001).The mean astigmatic magnitudes of KA and TCA were 1.44D×89.6° and 1.32 D×89.5° in astigmatism with the rule group,1.39 D×153.4° and 1.71 D×154.4° in astigmatism against the rule group and 1.13 D× 122.0° and 1.24 D× 124.2° in oblique astigmatism group.53 eyes (69.7％) had KA higher than TCA in astigmatism with the rule group.82 eyes (87.3％) had KA lower than TCA in astigmatism against the rule group;20 eyes (66.7％) had KA lower than TCA in oblique astigmatism group.There were significant differences in KP (0) between KA and TCA in different astigmatism groups (all at P＜0.001).The calculated Toric IOL type were inconsistent in 85 eyes(42.5％) and the calculated axis were inconsistent in 176 eye s (88.2％).Conclusions In patients with high corneal astigmatism,the astigmatism type of PA is mostly astigmatism against the rule.Ignoring the PA can lead to deviation of Toric IOL type selection and axis placement in some patients.For patients who cannot measure PA or TCA,the type of Toric IOL should be adjusted appropriately.