[Establishment and gene expression analysis of drug-resistant cell lines in hepatocellular carcinoma induced by sorafenib].

OBJECTIVE: To establish the drug-resistant cell lines of hepatocellular carcinoma (HCC) induced by sorafenib, and to screen out the high expression genes in drug-resistant cell lines of HCC induced by sorafenib, then to explore the genes related to sorafenib resistance in hepatocellular carcinoma. METHODS: The human PLC and Huh7 cell lines were obtained, then the PLC and Huh7 drug-resistant cell lines were induced with sorafenib by using intermittent induction in vitro. CCK8 assay was used to detect the IC50 value of sorafenib for evaluation of drug sensitivity of hepatocellular carcinoma cell lines in PLC and Huh7. All the up regulated genes in PLC and Huh7 drug-resistant cell lines induced by sorafenib were screened out using high-throughput cDNA sequencing (RNA-Seq), Ualcan database was used to analyze the correlations between the up regulated genes in PLC and Huh7 drug-resistant cell lines induced and four clinical biological characteristics of hepatocellular carcinoma, including the gene expressions between normal samples and tumor samples, tumor stage, tumor grade, and patient overall survival, to find the genes that might be involved in the mechanism of sorafenib resistance of hepatocellular carcinoma. RESULTS: All the up regulated genes detected by the using high-throughput cDNA sequencing (RNA-Seq) in PLC and Huh7 drug-resistant cell lines were further screened out by following conditions:(1) genes co-expressed in PLC and Huh7 drug-resistant cells induced by sorafenib, (2) the fold change was more than 4 times and the difference was statistically significant (P <0.05), the top 12 up regulated genes in PLC and Huh7 drug-resistant cell lines were found, which were TPSG1, CBX4, CLC, CLEC18C, LGI4, F2RL1, S100A6, HABP2, C15ORF48, ZG16, FOLH1, and EPCAM. Compared with the correlations between the twelve genes and the clinical biological characteristics by Ualcan database, the potentially significant gene CBX4 was screened out. CONCLUSION: The human PLC and Huh7 drug-resistant cell lines of hepatocellular carcinoma induced by sorafenib were successfully established. CBX4, the gene related to sorafenib resistance in hepatocellular carcinoma, was screened out by the high-throughput cDNA sequencing (RNA-Seq) and further analysis using Ualcan database, which is providing a powerful basis for further research on the mechanism of sorafenib resistance of hepatocellular carcinoma.

[Clinical features of long segmental myelopathy caused by nitrous oxide].

OBJECTIVE: To analyze the clinical and imaging characteristics of the neurological damage caused by nitrous oxide (N2O). METHODS: In the study, 10 patients in the Department of Neurology of China-Japan Friendship Hospital from October 2015 to February 2018 were retrospectively analyzed for the demographic data, the history of inhaled N2O, clinical features, blood examination, electrophysiological examination, spinal magnetic resonance imaging and therapeutic efficacy profiles. RESULTS: The male-to-female ratio was 4:6 and it presented with an age-of-onset 17-26 years [the average age: (20.80±3.12) years]. The time from inhaled N2O to onset was 1 month to 1 year [the average time: (6.95±4.19) months]. Paralysis in all the patients and numbness in 9 patients were the main clinical features, while positive Lhermitte's sign in 3 patients, urinary and defecation disturbance in 4 patients were also found. Blood examination indicated anemia in 2 patients, giant cell anemia in 1 case and small cell hypochromic anemia in 1 case. 3 cases had been treated with vitamin B12 in an external hospital, and the other 7 cases had abnormal increase in homocysteine levels. Electrophysiological examinations showed sensory and motor nerve involvement in 9 patients, and motor nerve involvement in 1 patient. The severity of lower extremity lesion was significantly heavier than that of upper extremity. Spinal magnetic resonance imagings showed that long segmental lesions were present in the cervical spinal cord of all the patients, 3 cases with long segmental lesions of the thoracic cord and 2 cases with spinal cord swelling. In 6 cases, the horizontal axis had an "inverted V-type" T2 high signal, 1 case was classified as "crescent", and 3 cases were "eight-shaped". The symptoms in these 10 cases were alleviated in varying degrees after stopping the inhalation of nitrous oxide, actively supplementing high doses of vitamin B12 and doing early rehabilitation exercises. CONCLUSION: Myelopathy with nitrous oxide presents as paralysis and numbness in limb extremities. In imaging, cervical spinal cord damage is common, accompanied by thoracic spinal cord damage. The horizontal axis is more common in the "inverted V-type". Treatment with high doses of vitamin B12 is effective.

[Analysis of population attributable risk of large for gestational age].

Objective: To examine the association of pre-pregnancy obesity, excessive gestational weight gain (GWG) and gestational diabetes mellitus (GDM) with the risk of large for gestational age (LGA), and assess the dynamic changes in population attributable risk percent (PAR%) for having these exposures. Methods: A retrospective cohort study was conducted to collect data on pregnant women who received regular health care and delivered in Beijing Obstetrics and Gynecology Hospital from January to December in 2011, 2014 and 2017, respectively. Information including baseline characteristics, metabolic indicators during pregnancy, pregnancy complications, and pregnancy outcomes were collected. Multivariate logistic regression model was constructed to assess their association with LGA delivery. Adjusted relative risk and prevalence of these factors were used to calculate PAR%and evaluate the comprehensive risk. Results: (1)The number of participants were 11 132, 13 167 and 4 973 in 2011, 2014 and 2017, respectively. Corresponding prevalence of LGA were 15.19% (1 691/11 132), 14.98% (1 973/13 167) and 16.21% (806/4 973). No significant change in the prevalence of LGA was observed across all years investigated (all P>0.05). (2)According to results from multivariate logistic regression model, advanced maternal age, multiparity, pre-pregnancy overweight or obesity, GWG,GDM and serum triglyceride level≥1.7 mmol/L in the first trimester were associated with high risk of LGA (all P<0.05). Among these factors, pre-pregnancy overweight or obesity, excessive GWG and multiparity were common risk factors of LGA. GDM was not associated with risk of LGA in 2017 database. (3) Dynamic change of PAR% in these years were notable. PAR% of GWG for LGA decreased (32.6%, 27.2% and 22.2% in 2011, 2014 and 2017, respectively), while PAR% of pre-pregnancy overweight or obesity showed an upward trend (4.2%, 3.3% and 8.4%). In addition, PAR% of multiparity increased as well (3.5%, 6.3% and 15.9%). (4) Further analysis showed that excessive GWG in the first and second trimesters contributed the most (20.2% and 19.0% in 2014 and 2017). Conclusions: Excessive GWG, pre-pregnancy overweight or obesity and multiparity are the important risk factors what contribute to LGA. PAR% of excessive GWG for LGA decrease in recent years. However, GWG in the first and second trimesters is a critical factor of LGA. Appropriate weight management in pre-pregnancy, the first or second trimester is the key point to reduce the risk of LGA.

Population genetic structure and migration patterns of Liriomyza sativae in China: moderate subdivision and no Bridgehead effect revealed by microsatellites.

While Liriomyza sativae (Diptera: Agromyzidae), an important invasive pest of ornamentals and vegetables has been found in China for the past two decades, few studies have focused on its genetics or route of invasive. In this study, we collected 288 L. sativae individuals across 12 provinces to explore its population genetic structure and migration patterns in China using seven microsatellites. We found relatively low levels of genetic diversity but moderate population genetic structure (0.05 < F ST < 0.15) in L. sativae from China. All populations deviated significantly from the Hardy-Weinberg equilibrium due to heterozygote deficiency. Molecular variance analysis revealed that more than 89% of variation was among samples within populations. A UPGMA dendrogram revealed that SH and GXNN populations formed one cluster separate from the other populations, which is in accordance with STRUCTURE and GENELAND analyses. A Mantel test indicated that genetic distance was not correlated to geographic distance (r = -0.0814, P = 0.7610), coupled with high levels of gene flow (M = 40.1-817.7), suggesting a possible anthropogenic influence on the spread of L. sativae in China and on the effect of hosts. The trend of asymmetrical gene flow was from southern to northern populations in general and did not exhibit a Bridgehead effect during the course of invasion, as can be seen by the low genetic diversity of southern populations.

Analysis of Fcgrt gene polymorphism in indigenous Chinese sheep and its association with colostrum IgG concentration.

The neonatal Fc receptor (FcRn) plays an important role in regulating IgG homeostasis in the body and passive protection to the offspring. Changes in FcRn expression levels caused by genetic polymorphisms of Fcgrt, which encodes FcRn, may lead to inter-individual differences in colostrum IgG levels in sheep. In this study, we sequenced the FcRn partial heavy chain from 179 sheep from Xinjiang Province, China, and detected the differences in colostrum IgG levels and Fcgrt genotypes to identify the correlation between the Fcgrt genotype and colostrum IgG levels in 4 sheep breeds. The DNA sequencing of a 680-bp fragment of the Fcgrt gene revealed various patterns depending on the single-strand conformation in the Suffolk breed. Sequencing analysis revealed a total of 3 patterns, AA, BB, AB, in this fragment, among which the absence of AB and BB genotype acted as a marker for breed identification and characterization, while the AA genotype was shared by Suffolk and 3 other breeds. The only allele found in all 4 breeds was allele A, indicating that natural selection may be favoring the AB and BB genotypes in general and B allele in particular, as the colostrum IgG concentration was relatively higher in the Suffolk breed compared to the other 3 breeds.

Expressions of Src homology 2 domain-containing phosphatase and its clinical significance in laryngeal carcinoma.

We investigated the expression of Src homology 2 domain-containing phosphatase (SHP-2) in laryngeal carcinoma and its clinical significance. Expression of SHP-2 was detected by immunohistochemical staining in normal mucosal tissues and various grades of laryngeal carcinoma. We looked for possible correlations between expression of SHP-2 in laryngeal carcinoma and clinical staging and lymph node metastasis. Immunochemical staining results revealed that the SHP-2 expression was significantly higher (88.24%) in laryngeal carcinoma than in normal mucosal tissue (25%). Additionally, the expression of SHP-2 was significantly correlated with lymph node metastasis, but not with clinical stage and gender of patients with laryngeal carcinoma. Therefore, SHP-2 may be useful as a prognostic marker for laryngeal carcinoma and as a therapeutic target in laryngeal carcinoma treatment.

Long-term dietary exposure to lead of the population of Jiangsu Province, China.

Deterministic and probabilistic estimates of the chronic exposure to lead (Pb) for the inhabitants in Jiangsu Province, China, were performed. Pb contamination data were obtained from the national food contamination monitoring programme during 2001-2006 and 2600 samples from 38 commodities in Jiangsu were included. Food consumption data were from the national diet and nutrition survey conducted in 2002, including 3879 subjects aged 2-80 years and 185 children aged 2-6 years in Jiangsu. Contributions from 38 commodities were included in the calculations. Using the provisional tolerable weekly intakes (PTWI) divided into daily intakes (PTWI(d)) of 3.57 microg kg(-1) bw day(-1) in the risk assessment, exposures calculated by the deterministic approach for children and the general population were 77% and 43% of the PTWI(d), respectively, which were nearly similar to the mean intake calculation in the probabilistic approach. The percentage of people whose exposure level exceeded the PTWI(d) for children, the general population, urban population and rural population were 16%, 0.4%, 0.1%, and 0.5%, respectively. The 50th, 95th and 99th percentiles of Pb exposure for children were 2.6, 4.4 and 5.5 microg kg(-1) bw day(-1), respectively. Rice followed by wheat flour and bean products contributed most to the total Pb intake for both children and the general population. This study indicated that the Pb dietary exposure in Jiangsu Province, especially for children, would be a public health concern.