Objective To assess the prognostic value of KRAS mutation in patients with advanced primary liver cancer treated with transcatheter arterial chemoembolization (TACE). Methods Ninety-seven patients with advanced primary liver cancer who received TACE treatment in The Third People's hospital from April 2017 to May 2020 were included. The mutation status of KRAS was detected, and its relationship with the prognosis of TACE was investigated. The t -test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. Survival analysis was performed using Kaplan-Meier survival curve and compared using Log-rank test. Cox regression analysis was performed to identify the prognostic factors. Results Among 97 patients with advanced liver cancer, KRAS mutations were detected in 34 patients (35.05%), including 21 patients with codon 12 mutation (61.76%) and 13 patients with codon 13 mutation (38.24%). KRAS mutation was associated with liver cirrhosis, intrahepatic metastasis and the number of tumors ( χ 2 =0.035, 3.965, and 6.593, all P < 0.05). Survival analysis showed that the progression free survival and overall survival were significantly longer in KRAS wild-type patients than in KRAS mutant patients ( χ 2 =4.465 and 4.280, all P < 0.05). Multivariate Cox analysis revealed that KRAS mutation, intrahepatic metastasis, number of tumors and BCLC stage were important factors affecting the overall survival and prognosis of patients (all P < 0.05). Conclusion KRAS mutation is common in patients with advanced primary liver cancer and is closely associated with a poor prognosis after TACE. It may become a potential indicator of clinical prognosis.

Objective:To investigate the effect of virus inactivation on weak positive result of 2019 novel coronavirus(2019-nCoV) nucleic acid test.Methods:A retrospective study was conducted on the nasopharyngeal swabs of three patients with positive PCR nucleic acid test for 2019-nCoV at different concentrations in the Second affiliated Hospital of Zhejiang University Medical College from January to February 2020.The virus in nasopharyngeal swab specimens were inactivated by water bath at 56 ℃ for 30 min, dry bath at 56 ℃ for 60 min and dry bath at 60 ℃ for 30 min respectively. After treatment, these samples RNA were extracted and then detected by three new commercial quantitative real-time polymerase chain reaction reagent kits for 2019-nCoV.Cycle threshold (Ct) value was used to evaluate the effect of virus inactivation on nucleic acid detection of 2019-nCoV.Results:There was no significant difference between the groups before and after inactivation. Ct values of ORF1ab gene before inactivation were 23.28±0.28, 25.25±0.25, 28.93±0.44, 32.06±0.47, 35.20±0.38, 32.89±0.38, 36.24±0.23, 33.30±0.46, and those after inactivation were, group 1:23.60±0.20, 27.29±0.30, 31.83±0.51, 37.41±0.46, group 2: 24.25±0.34, 27.18±0.42, 31.84±0.61, 34.99±1.01, 34.89±0.45,group 3: 23.37±0.17, 26.89±0.52, 32.05±0.50.Ct value of N gene before inactivation were 24.38±0.09, 26.64±0.11, 30.35±0.12, 33.29±0.33, 36.93±0.11, 34.50±0.12, 35.63±0.12, those after inactivation were, group 1: 24.66±0.11, 28.52±0.14, 32.71±0.14, 37.00±0.13;group 2: 25.41±0.10, 28.79±0.15, 33.29±0.28; group 3: 23.37±0.11, 28.68±0.11, 33.54±0.13, 37.18±0.23(ORF1ab gene: t=-1.416; N gene: t=-1.379, P>0.05). There was no significant difference among the three inactivation groups, the specific Ct values are shown above(ORF1ab gene: t=-0.460; N gene: t=-0.132, P>0.05). However, the Ct values of the inactivated groups (1,2,3) and the non-inactivated group at different dilution times were different (10 ×:Ct value of ORF1ab was 25.25±0.25 in the non-inactivated group, and 27.29±0.30, 27.18±0.42 and 26.89±0.52 in the inactivated group1,2 and 3, t(ORF1ab)=-7.327, P<0.01.Ct value of N gene in the non-inactivated group was26.64±0.11, those in inactivated group 1, 2 and 3 were 28.52±0.14, 28.79±0.15 and 28.68±0.11, respectively, t (N)=-19.340, P<0.01. 100 ×:Ct value of ORF1ab was 28.93±0.44 in the non-inactivated group, and 31.83±0.51,31.84±0.61 and 32.05±0.50 in the inactivated group1,2 and 3, t (ORF1ab)=-9.462, P<0.01. Ct value of N gene in the non-inactivated group was 30.35±0.12, those in the inactivated group 1, 2 and 3 were 32.71±0.14, 33.29±0.28 and 33.54±0.13, respectively, t (N)=-18.583, P<0.01. The positive detection rate of the non-inactivated group (7/11, 8/11, 5/11) was significantly different from that of the inactivated group (inactivated group 1:4/11, 4/11, 3/11, inactivated group 2:3/11, 3/11, 3/11, and inactivated group 3:3/11, 3/11, 2/11) ( Z=-2.670, P<0.01). There were no significant difference among the inactivated groups(inactivated group 1:4/11, 4/11, 3/11, inactivated group 2:3/11, 3/11, 3/11, inactivated group 3:3/11, 3/11, 2/11) ( Z=4.413, P>0.05) and among the three reagents(reagent 1:7/11, 4/11, 3/11, 3/11, reagent 2:8/11, 4/11, 3/11, 3/11, reagent 3:5/11, 3/11, 3/11, 2/11)(χ 2=1.199, P>0.05). Conclusion:The virus inactivation can degrade the nucleic acid of the 2019-nCoV, resulting in the decrease of the Ct value and the false negative results of the low-concentration specimens.

Objective To investigate the characteristics of metabolic foci on 18F-fluorodeoxyglucose (FDG) PET/CT scan in pediatric patients with epilepsy.Methods Twenty-three pediatric patients (15 males,8 females,age range:0.5-13.3 years) with epilepsy were retrospectively reviewed from March 2014 to December 2016.All patients underwent 18F-FDG PET/CT and metabolic foci were found.The visual method and semi-quantitative analysis were used to analyze images.Fourteen of them underwent surgery and were followed up for 3-24 months.Results Glucose hypermetabolism were observed most frequently in the frontal and parietal lobes,with or without surrounding/remote hypometabolism.On the day of PET/CT imaging,8 patients had no seizures,14 patients had seizures,and 1 patient was uncertain.The sites of resection were consistent with the regions of hypermetabolism in 9 patients,among whom the pathological results showed 8 cortical malformations and 1 Rasmussen's syndrome.Follow-up results for the above 9 patients showed that there was 7 Engel class Ⅰ patients and 2 Engel class Ⅲ patients.Conclusion The hypermetabolism may mostly appear in the frontal and parietal lobes of pediatric patients with epilepsy,and malformations of cortical development seem to be the most common pathology results.

Objective To investigate the clinical value of 18 F?FDG PET/CT in staging multiple myeloma ( MM) and evaluating the glucose metabolic activity of MM. Methods A total of 25 MM patients ( 13 males, 12 females, age:39-67 years) from May 2010 to April 2015 were enrolled in this retrospective study. The SUVmax of each patient was recorded. D?S plus staging according to 18 F?FDG PET/CT was com?pared with the traditional D?S staging. The SUVmax and the percentage of plasmacytes of bone marrow of phase Ⅲ and non?phase Ⅲ ( phaseⅠand Ⅱ) according to D?S plus staging were compared. Two?sample t test and Wilcoxon rank sum test were used to analyze the data. Results In 25 MM patients, the range of SUVmax of lesions was 1.8-12?0 and the mean value was 5.15±2.74. According to D?S staging, the numbers of patients with phase Ⅰ,Ⅱ andⅢwere 7, 4 and 14, respectively. While the numbers were 3, 1 and 21 by D?S plus staging. Based on the D?S plus staging system, stages of 7 patients ( 28%, 7/25 ) were changed. According to the D?S plus staging system, the SUVmax between phaseⅢand non?phaseⅢpatients was significantly different (5.75±2.54 vs 3.00±0?70; t=2.12, P<0.05), while the percentage of plasma?cytes of bone marrow between the 2 groups had no significant difference ( 17. 50%( 4. 25%-41. 75%) vs 11?15%(10.25%-36.57%);z=0.05, P>0.05). Conclusion 18F?FDG PET/CT is of clinical importance for MM staging and metabolic activity assessment of MM.

Objective To investigate the levels of blood pressure, blood lipid, blood glucose, body mass index (BMI) as well as the epidemiological characteristics of dyslipidemia, hyperglycemia, hypertension and metabolic syndrome of Hangzhou citizens. Methods A total of 28 990 citizens in Hangzhou city who underwent health checkup were recruited in this study, including 10 179 males and 18 811 females. The average age of subjects was 65.05 years. Subjects were asked to complete questionnaires regarding personal characteristics. The physical examination emphasized measurement of height, waist and blood pressure. Blood samples were collected and subjected to serum glucose, TC, HDL-C, LDL-C and TG measurements. The values of the examinations was described as xˉ± s . The ratios were compared with chi-square test. The trend analysis was conducted by linear correlation test. Results The prevalence of hypertension and metabolic syndrome was 17.1% and 11.2% respectively. And the prevalence of overweight/obesity and hyperglycemia was 36.3%,8.1%,16.4%respectively. It was indicated that the men had higher prevalence of hyperglycemia, hypertension, metabolic syndrome and overweight compared with women. However, as to the dyslipidemia, men and women were totally different. Women were more prone to suffer from hypercholesterolemia and elevation of low-density lipoprotein cholesterol. Men were apt to suffer from hypertriglyceridemia and reduction of high-density lipoprotein cholesterol. Divided the subjects by age into three groups, it was suggested that the rates of dyslipidemia, hyperglycemia, hypertension and overweight/obesity increased along with the increment of age in women. Although the rates of metabolic disorders were higher in the group of men, the trend of increase with age was not as significant as in women. It could be seen in men that dyslipidemia and overweight/obesity were reduced with the increase of age. Conclusion The metabolic disorders in Hangzhou citizens showed their own characteristics. It is suggested that multiple strategies targeting at different sexes and age-groups should be formulated to prevent and control the occurrence of metabolic diseases.

Objective In order to assess the value of Echinoccocus granulosus cyclophilin A (EgypA) in immune diagnosis,this novel gene was cloned and expressed.Methods By screening the EST library,the coding region of EgCypA was identified,and the PCR primers were designed based on this sequence.Bioinformatic tools were used to deduce the amino acid sequence of EgCypA and analyzed its biological characteristics.EgCypA was amplified from E.granulosus cDNA library by using PCR.Then,it was cloned into the prokaryotic expression vector pET28a and transformed into E.coli BL21.The recombinant protein was expressed in E.coli BL21 after IPTG induction.The immunogenicity of EgCypA was evaluated by Western blot using the Echinoccocus granulosus and other parasites infected animals' sera.Results Results of SDS-PAGE electrophoresis show that the recombinant BL21 expressed 18 400-25 000 protein,which was identical with the molecular weight calculated by bioinformatic analysis.Western blot shows that the recombinant protein only reacted with its immune serum and E.granulosus cystic fluid immune serum,and EgCypA immune serum could react with the excretion and secretion antigen from E.granulosus protoscolexes,and no cross-reaction between EgCypA and sera from other parasites infected animals.Conclusions Cloning and expression of EgCypA are successful.EgCypA has good immunologic activity and could be a candidate molecular for immune diagnosis of echinococcosis in early stage.

Despite its clinical importance, the underlying central mechanisms of pruritic behaviors are poorly understood. To investigate the role of nociceptive arcuate nucleus neurons in chloroquine-induced pruritic behaviors in mice, we tested the effect of arcuate nucleus neurons and interscapular brown adipose tissue (IBAT) on itch produced by intradermal injection of chloroquine in the nape of the neck. Our results provide several lines of evidence for an important role of nociceptive arcuate nucleus neurons in chloroquine-induced pruritic behavior: (1) Intradermal microinjection of chloroquine resulted in a dramatic increase in itch behaviors accompanied by the activation of c-Fos positive neurons in arcuate nucleus; (2) Microinjection of chloroquine significantly increased IBAT temperature in the mice. These findings suggested that chloroquine-induced pruritic behaviors were associated with the activity of nociceptive arcuate nucleus neurons.
Animals ; Arcuate Nucleus of Hypothalamus ; drug effects ; physiopathology ; Chloroquine ; adverse effects ; Male ; Mice ; Mice, Inbred C57BL ; Nociceptors ; drug effects ; physiology ; Pruritus ; chemically induced ; physiopathology

Despite its clinical importance, the underlying central mechanisms of pruritic behaviors are poorly understood. To investigate the role of nociceptive arcuate nucleus neurons in chloroquine-induced pruritic behaviors in mice, we tested the effect of arcuate nucleus neurons and interscapular brown adipose tissue (IBAT) on itch produced by intradermal injection of chloroquine in the nape of the neck. Our results provide several lines of evidence for an important role of nociceptive arcuate nucleus neurons in chloroquine-induced pruritic behavior: (1) Intradermal microinjection of chloroquine resulted in a dramatic increase in itch behaviors accompanied by the activation of c-Fos positive neurons in arcuate nucleus; (2) Microinjection of chloroquine significantly increased IBAT temperature in the mice. These findings suggested that chloroquine-induced pruritic behaviors were associated with the activity of nociceptive arcuate nucleus neurons.