Objective To explore the impact of iodine nutrition on 8-10 years old children after adjusting the iodine content in iodized salt in Hangzhou.Methods Twelve counties (areas,cities) were divided into urban,suburban and rural areas in Hangzhou.By population proportion survey (PPS),every county(area,city) was divided into east,west,south,north and middle districts; one school was selected in each district; forty children (half male and half female) aged 8-10 years old in each school were selected; family salt and urine samples of each student were collected.The levels of salt and urinary iodine were measured by picric sodium thiosulfate titrimetric (GB 13025.7-2012) and spectrophotometer method (WS/T 107-2006),respectively.Results Two thousand seven hundred and twenty-five household salt samples were collected.The median of salt iodine,the iodized salt coverage rate,the qualification rate of iodized salt and the consumption rate of qualified iodized salt were 24.00 mg/kg,4.35％(2 571/2 725),91.02％(2 340/2 571) and 85.87％(2 340/2 725),respectively.The medians of salt iodine in urban,suburb and rural areas were 24.10,22.12,24.30 mg/kg,respectively.A total of 2 664 children urine samples were collected.The median of urinary iodine (MUI) of the children was 177.24 μg/L.The MUIs in urban,suburb and rural areas were 175.00,178.55,178.00 μg/L,respectively; in male was 183.00 μg/L and female was 170.50 μg/L.When non-iodized and unqualified iodized salt were taken,the differences of urinary iodine within groups were statistically significant in urban,suburb and rural areas(x2 =18.652,14.686,all P ＜ 0.05).In rural area,the difference of urinary iodine of 8-10 years old children who ingested different types of iodized salt was statistically significant(x2 =39.07,P ＜ 0.05).Conclusion After adjusting the iodine content of salt in Hangzhou,the iodine-nutritional status of 8-10 years old students is at a appropriatelevel.

Objective: : To explore the effect of PD-1 gene knockout by CRISPR/Cas9 system on the proliferation and IFN-γ secretion in human T cells. Methods: : The sequence of sgRNA targeting PD-1 was designed. The PD-1-sgRNA and Cas9 mRNA were synthesized by T7 RNApolymerase in vitro, and then the mixture of PD-1-sgRNAand Cas9 mRNAwas delivered into activated T cells by nucleofection. The efficiency of gene knockout was confirmed by sequencing. The phenotypes of T lymphocytes and the expression of PD-1 after gene knockout were analyzed by Flow cytometry. The proliferation of T lymphocytes was calculated by trypan blue counting. The level of IFN-γ secreted by T lymphocytes was detected by ELISA. Results： ：PD-1-sgRNA and Cas9 mRNA were successfully synthesized in vitro and delivered into T cells by nucleofection. Sequencing technology confirmed that the PD-1 gene sequence was edited and the editing efficiency was 58.3%. The expression of PD-1 on T lymphocyte surface was down-regulated successfully by CRISPR/Cas9 system [(9.6±1.85)% vs (16.2±2.05)%, P<0.05]. The knockout of PD-1 gene did not affect the proliferation and phenotype of T lymphocytes(P<0.05); However, compared with the control group, the level of IFN-γ secreted by T lymphocytes in the PD-1sgRNA group was significantly increased (P<0.01). Conclusion: : CRISPR/Cas9 system can successfully ablate PD-1 gene in human T lymphocytes, which could block the negative regulation of PD-1／PD-L1 and further promote the IFN-γ secretion in T cells.

Objective: To understand the changing trend and influencing factors of health literacy among residents in Hangzhou from 2016 to 2020, so as to provide basis for health literacy promotion. Methods: The monitoring data of health literacy of residents in Hangzhou from 2016 to 2020 were collected. The five year cumulative growth and average growth rate were used to reflect the changing trend. The multivariate logistic regression model under complex survey design was used to analyze the influencing factors for health literacy. Results: From 2016 to 2020, the health literacy levels of Hangzhou residents were 26.91%, 29.49%, 31.83%, 34.73% and 38.00%, respectively. The five year cumulative growth was 11.09% and the average growth rate was 9.01%. Among three aspects of health literacy, the levels of basic knowledge and concept as well as healthy lifestyle and behaviors increased year by year (P<0.05), with the average growth rates of 15.12% and 2.92%. Among six types of health problems, the levels of chronic disease prevention and treatment, basic medical treatment, scientific view, health information as well as safety and first aid increased year by year ( P<0.05 ), with the average growth rates of 32.76%, 15.82%, 8.41%, 7.12% and 1.53%, respectively. The multivariate logistic regression analysis showed that education level ( junior high school, OR=5.359, 95%CI: 1.151-24.953;high school/vocational high school/technical secondary school, OR=9.214, 95%CI: 2.906-29.213; college or above, OR=29.977, 95%CI: 9.689-92.741 ) and occupation ( students, OR=2.564, 95%CI: 1.113-5.907 ) were the influencing factors for health literacy. Conclusion The health literacy levels of Hangzhou residents from 2016 to 2020 have been increasing year by year. The residents with higher education levels and students may have higher health literacy levels.

Objective: To learn the current situation of smoking among permanent residents aged 15 years and over in Hangzhou,so as to provide evidence for further smoking control. Methods: From March to June of 2017,multi-stage random sampling method was used to select residents aged 15 years and over in Hangzhou from district (county),town (street),village (residential committee) and then residential groups. The global Adult Tobacco Survey (part of China) questionnaire was used to investigate smoking,second-hand smoking and smoking cessation status of residents. Logistic regression model was used to analyze the influencing factors for smoking and quitting behaviors. Results: A total of 1 440 questionnaires were sent out and 1 434 valid ones were recovered,with an response rate of 99.58%. There were 359 (25.03%) current smokers and 333 (23.22%) daily smokers. The median number was 15 cigarettes per day in daily smokers,and the median age of starting daily smoking was 21 years old. There were 670 (62.33%) people exposed to second-hand smoking. There were 137 (27.62%) people successfully quitting smoking. The results of multivariate logistic regression analysis showed that men (OR=73.459,95%CI:38.970-138.458), 50 years old or above (OR:1.646-2.632,95%CI:1.020-4.710),primary school education (OR=3.267,95%CI: 1.586-6.732),high school or vocational school education (OR=1.902,95%CI: 1.109-3.261),awareness of second-hand smoking hazards (OR=2.485,95%CI:1.735-3.559) were the influencing factors for current smoking;men (OR=64.454,95%CI:29.031-138.691),50-59 years old (OR=0.281,95%CI:0.095-0.827),primary school education (OR=0.211,95%CI:0.053-0.841) were the influencing factors for quitting smoking. Conclusions The rates of current smoking and second-hand smoking of permanent residents aged 15 years and over in Hangzhou were 25.03% and 62.33%,respectively. Gender,age and education level were associated with current smoking and quitting smoking.

When we try to establish the gene recombinant yeast cell to screen the androgenic endocrine disruptors, the key procedure is the androgen receptor (AR) expression in the yeast cell. For this purpose, we obtained the GPD (glyceraldehyde-3-phosphote dehydrogenase) promoter from the yeast genosome of W303-1A using PCR system and inserting it into Swa I and BamH I sites of pYestrp2. The new constructed vector was named pGPD. The V5 epitope tag DNA with a 5'-BamH I and a 3'-EcoR I sticky end was cloned into the corresponding site of the pGPD vector to yield the vector of pGPDV5. The 2 723 bp full length AR ORF amplified by PCR from pcDNA3.1/AR was fused to V5 epitope tag DNA in pGPDV5 to give the AR yeast expression vector of pGPDV5/AR. This fused vector was transformed into the yeast cell (W303-1A). Western blot was used to detect the V5 fused protein of AR, in the protocol of which the primary monoclonal antibody (IgG(2a)) of mouse anti-V5 and the polyclonal secondary antibody of goat anti-mouse (IgG) linked to horseradish peroxidase (HRP) were used to detect the specific protein in the given sample of the transformed yeast extract. The result showed that the fused protein of AR was expressed successfully in the yeast cell.
Base Sequence ; Endocrine Disruptors ; analysis ; Epitopes ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Glyceraldehyde-3-Phosphate Dehydrogenases ; genetics ; Humans ; Molecular Sequence Data ; Promoter Regions, Genetic ; Receptors, Androgen ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; genetics ; Yeasts ; genetics ; metabolism

Objective: To analyze knowledge, attitudes and practice regarding protection against COVID-19 among primary and middle school students in Hangzhou and to provide a scientific basis for schools to carry out targeted prevention and control measures. Methods: Anonymous network sampling survey was carried out through the questionnaire star, and a total of 14 216 subjects from primary and middle school in Hangzhou were recruited to fill in the questionnaire regarding knowledge, attitudes and practice of COVID-19 from March 3 to March 7, 2020. Results: Primary and middle school students showed high awareness rate of transmission (85.83%) and incubation period (77.44%), but lower awareness on symptoms (44.70%) of COVID-19. The practice of wearing masks, cough etiquette and correct hand washing were 97.27%, 88.01% and 91.82%, respectively. The protective practice among primary and middle school students were generally good, 91.21% of them were at moderate level or above. 83.91% of primary and secondary school students reported worries about infection of COVID-19, and 55.42% of primary and secondary school students were anxious about returning to school. Primary school students and rural students were more anxious about returning to school(χ 2=46.66, 26.70, P<0.05). Conclusion The primary and middle school students in Hangzhou have a relatively good level of knowledge and practice concerning COVID-19. Yet, it is necessary to carry out targeted health education and strengthen mental health education for primary and middle school students.

OBJECTIVETo construct γ-synuclein gene eukaryotic expression vector, and to study its effect on the invasion of colon cancer cell line SW1116 and the adhesion between SW1116 and human umbilical vein endothelial cells(HUVECs) in vitro.

METHODSTotal RNA was extracted from colon cancer cell line HT29 and the cDNA of γ-synuclein was amplified using RT-PCR. The digested fragment of cDNA coding sequence was linked to the eukaryotic expression vector pEGFP-C1 containing the GFP gene. After identification by sequence analysis, the recombinant plasmid was transfected into colon cancer cell line SW1116 via lipofectamine. The stable cell line was selected with G-418. The invasion in vitro was tested by Transwell invasion chamber assay. HUVECs were previously seeded onto 96-well plates before SW1116 cells seeded, and fluorescence intensity of GFP was detected to represent the amount of adhesion cells by ELISA.

RESULTSHuman γ-synuclein eukaryotic expression vector was successfully constructed, which was stably expressed in SW1116 cells and could translate the GFP-γ-synuclein protein in vitro. γ-synuclein facilitated SW1116 cell passing through matrigel and filter membrane(198.4±20.7 vs. 98.8±13.2, P<0.05) and elevated the adherence of SW1116 cells to HUVECs(3.08±0.36 vs. 1.22±0.21, P<0.05).

CONCLUSIONExpression of γ-synuclein can strengthen colon cancer cell SW1116 potentiality of invasion and metastasis in vitro.

Cell Adhesion ; Cell Line, Tumor ; Colonic Neoplasms ; pathology ; Genetic Vectors ; biosynthesis ; Human Umbilical Vein Endothelial Cells ; Humans ; Neoplasm Invasiveness ; Neoplasm Metastasis ; gamma-Synuclein ; genetics

OBJECTIVETo construct a lentiviral vector carrying the γ-synuclein(SNCG) gene and establish a human colorectal carcinoma cell line SW1116 stably expressing this gene, and then investigate the inhibition of the growth and invasion capacity of SW1116 cells.

METHODSRNA interference fragment was designed according to the SNCG sequence (GenBank: No.NM003087.2), and then SNCG RNAi effective target genes were screened. After the Oligo DNA of target sequences was synthesized, the lentiviral vectors carrying LV-SNCG-RNAi-EGFP (RNAi group) and LV-SNCG-NC-EGFP (NC group) were constructed and packaged to produce lentivirus venom. The supernatants of different virus-producing cells were used to transfect SW1116 cells respectively. Wild SW1116 cells were used as blank control (CON group) EGFP fluorescence was detected by fluorescent microscopy and the differential expression of SNCG mRNA and protein was detected by real-time PCR and Western blot. CCK-8, soft agar assay and Transwell chamber were employed to estimate the inhibiting effect on growth and invasion of SW1116 respectively.

RESULTSRecombinant lentiviral vectors respectively carrying the SNCG-RNAi-EGFP and SNCG-NC-EGFP were successfully constructed and the supernatants of lentivirus could effectively infect SW1116 cells. The titer of the virus carrying LV-SNCG-RNAi-EGFP or LV-SNCG-NC-EGFP was 8×10(8) TU/ml. Real-time PCR and Western blot confirmed that compared with the NC group, SNCG-RNAi group had lower SNCG expression (1.009±0.161 vs. 0.114±0.030, P=0.009), and showed tremendous silencing effect as 76.8%(P<0.05). SNCG protein expression was also significantly reduced (RNAi:12.001±2.884, NC:32.443±4.731, CON:34.308±6.920, P<0.05). After SNCG knockdown, the number of proliferation cells was obviously reduced at 48, 72, 96 and 120 hours respectively(P=0.036). In soft agar assay, clones in RNAi group were smaller[RNAi:(0.582±0.103) mm, NC:(1.863±0.316) mm, CON:(1.749±0.525) mm]. Colony formation rate of RNAi group was down to (17.1±3.5)%, which was significantly lower than (36.5±4.3)% in NC group and (33.8±3.9)% in CON group. In migration test, the number of invasion cell was 37.4±9.3 in RNAi group, which was significantly less than 112.3±8.6 in NC group and 100±0.0 in CON group.

CONCLUSIONExpression of SNCG mRNA and protein plays an important role in the growth and the invasion capacity of SW1116 cells.

Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; pathology ; Genetic Vectors ; Humans ; Lentivirus ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Transfection ; gamma-Synuclein ; genetics

Objective:To investigate the efficacy and safety of dual wavelength pulsed dye laser combined with 30% supramolecular salicylic acid in the treatment of moderate and severe facial acne.Methods:Sixty patients with moderate and severe acne that visited the Dermatology Department of the First Affiliated Hospital of Chongqing Medical University from May 2020 to January 2021, were selected and randomly divided into observation group and control group, with 30 patients in each group. The observation group was given dual-wavelength pulsed dye laser combined with 30% supramolecular salicylic acid. 30% supramolecular salicylic acid was used once every two weeks, for a total of six times. Dual-wavelength pulsed dye laser was given once a month, for a total of three times. The control group was only given dual-wavelength pulsed dye laser, once a month, a total of three times.Results:Twenty-two cases (73.33%) in the observation group were effective, while 14 cases (46.67%) in the control group were effective. The efficacy of the observation group was better than that of the control group, and the difference was statistically significant (χ 2=4.44, P<0.05). There were no obvious adverse reactions in both groups. Conclusions:Dual-wavelength pulsed dye laser combined with 30% supramolecular salicylic acid is effective and safe in the treatment of moderate and severe facial acne, which is worth popularizing.