Objective To construct a luciferase reporter gene vector of perforin promoter and methylate it in vitro. Methods The promoter of the human peffofin was amplified by PCR, cloned into pMD18-T vector and subcloned into pGL3-Basie vector, and then con-finned by restriction mapping and DNA sequencing. The regions of interest were excised with the appropriate restriction endonucleases, then it were methylated with methylase Sss I(M. SssI)and S-adenosymethioine(SAM), and methylation was confirmed by digestion with appropri-ate methylation sensitive enzyme AciI and agrose gel electrophoresis, and then the fragment was relegated back into the promoter-reporter constructor pGL3-Basic. Results The result of DNA sequencing showed that the sequence of cloned promoter was right. The result of diges-tion methylation with appropriate methylation sensitive enzyme showed that perforin promoter was completely methylated. Conclusion The promoter of perforin was successfully cloned and completely methylated in vitro, which provided an important basis for the study of transfec-tion.

Objective:To analyze the monitoring results of iodine deficiency disorders in Nanning City in 2020, learn about the consumption of iodized salt among residents and the iodine nutrition status of key populations, and provide scientific basis for formulating or adjusting targeted prevention and control measures of iodine deficiency disorders.Methods:According to the Guangxi Iodine Deficiency Disorders Monitoring Plan, monitoring was carried out in all of 12 districts (counties) in Nanning City. One township (street) was selected from each of the five directions of east, west, south, north, and central. Forty non-boarding children aged 8 to 10 and 20 pregnant women were selected as monitoring subjects in each township (street). Edible salt samples were collected from children and pregnant women to detect salt iodine content, random mid-course urine samples from children and morning urine samples from pregnant women were collected to detect urinary iodine content; in addition, thyroid examination of children was conducted in Qingxiu District, Liangqing District, Long an County and Shanglin County.Results:In 2020, a total of 2 434 children aged 8 to 10 and 1 207 pregnant women were surveyed in Nanning City. The coverage rate of iodized salt, the qualified rate of iodized salt and the qualified iodine salt consumption rate were 99.67%(3 629/3 641), 97.99%(3 556/3 629) and 97.67%(3 556/3 641), respectively. The median urinary iodine of children was 182.0 μg/L, and the median values ranged from 146.5 to 234.8 μg/L in different districts (counties), there were significant differences in median urinary iodine between urban and non-urban areas, different gender and age groups ( U = 2.38, 2.41, P = 0.017, 0.016; H = 16.42, P < 0.001). The goiter rate of children was 0.99%(8/807), and the rate ranged from 0 to 2.00% (4/200) in the 4 districts (counties) examined, there were significant differences in thyroid volume between urban and non-urban areas and different ages ( U = - 3.52, P < 0.001; H = 47.67, P < 0.001). The median urinary iodine of pregnant women was 191.0 μg/L. The median urinary iodine of different districts (counties) ranged from 141.0 to 241.5 μg/L, and the difference was statistically significant at different gestational stages ( H = 24.37, P < 0.001). Conclusion:In 2020, the coverage rate of iodized salt, the qualified rate of iodized salt and the qualified iodine salt consumption rate by residents in Nanning City are high, and the iodine nutrition of both children and pregnant women are at an appropriate level.

This article introduces the contents of the latest edition Risk of Bias in Non-randomized Studies-of Exposure (ROBINS-E) published in June 2022 [ROBINS-E (2022)], and gives some examples about its usage. ROBINS-E is a tool for assessing the risk of bias in non-randomized studies-of exposure. Compared with ROBINS-E (2019), ROBINS-E (2022) adds more bias for observational studies, covers a more comprehensive range of bias, and adds the assessment of the external authenticity of the study. ROBINS-E (2022) adds a preliminary evaluation process to improve the efficiency of evaluation. In addition, ROBINS-E (2022) visualizes and instrumentalizes the use of signal problems in the form of path graph, making it more convenient to use. ROBINS-E (2022), although more consideration has been given to the issue of co-exposure, still does not address the problem of effect modification in co-exposure, and there is still room to expand the applicable research.

Objective To analysis the change of hip joint in healthy infants by ultrasound,and establish the normal reference value of the developmental dysplasia of the hip(DDH). Methods A total of 8 000 healthy infants from 0 to 24 weeks were collected from the Multi-center study of 10 children′s medical centers. Among them,3 855 infants(2 065 females and 1 790 males) with complete data and follow-up were included in this study. All subjects were divided into 6 groups ( ＜4,4～7,8～11,12～15,16～19 and≥20 weeks group). α angle,femoral head length and width,femoral head coverage ratio by acetabulum ( FHC) were measured in the coronal view on the neutral position;distance from pubis to femoral head ( P-H) and distance from ischium to femoral head ( I-H ) were measured in the transverse view on neutral position;distance from femoral head topubis ( H-P) was measured in the posterolateraltransverse view on the flexion position. The results of each group changes with age were analysised. Results ① The α angle of healthy infants from 0 to 20 weeks were increased with age,the difference among the groups were statistically significant( P ＜0.05),but there was no significant difference between 16～19 and ≥20 weeks group( P ＞0.05). ②The femoral head length and width of all age groups were increased with age,the difference among all the groups was statistically significant( all P ＜0.05). ③ FHC from 0 to 20 weeks were increased with age,the difference among the groups were statistically significant( P ＜0.05) except between 16～19 and ≥20 weeks group( P ＞0.05). ④ The P-H and I-H in all age groups showed no statistically significant ( all P＞0.05). ⑤The H-P of all age groups were increased with age,the difference between the groups were statistically significant(all P ＜0.05).Conclusions The development of hip joints have the certain regular developmental pattern in healthy infants less than 5 months of birth and are relatively constant after birth more than 5 months. The ultrasound normal reference value of the hip joints can be used for the early diagnosis of the DDH.