OCCURRENCE OF METFORMIN IN ENVIRONMENTAL WATER SAMPLES AND COMPARISON WITH CONSUMPTION DATA FROM A SURROUNDING HOSPITAL OVER 5 YEARS: A RETROSPECTIVE CASE STUDY.

Context: The environmental occurrence of metformin has been frequently world-widely reported. Despite the diabetes susceptibility in the Chinese population, the studies on occurrence of metformin as environment disruptor in China are insufficient. Objective: To determine the occurrence trends and possible environmental pollution sources of metformin as an emerging micropollutant. Methods: High-performance liquid chromatography-electrospray ionization-tandem mass spectrometry system was used to detect the metformin levels in water samples collected from the Xi'an city Moat, China once a year from 2017 to 2021. Correlations among the metformin levels in moat water, in surrounding hospital wastewater, and hospital metformin consumption data were assessed using Pearson, Spearman and Kendall's tau-b correlation coefficients. Results: Occurrence of metformin was found in Xi'an city Moat water with levels in the range of 304-793 ng/L. Significant correlations were found between the metformin levels in city moat water and the total (or outpatient) metformin utilization data of the hospital. Conclusion: Data suggested the potential environmental issues posed by metformin in Xi'an city in China. The metformin consumption volume in the surrounding hospitals, especially at the outpatient services, could be used to predict the metformin concentrations in the moat water.

Transcriptome analysis of the bursa of Fabricius and thymus of laying ducks reveals immune gene expression changes underlying the impacts of stocking densities.

1. The thymus and bursa of Fabricius are important immune organs in poultry as they play essential roles in sustaining the normal immune function to maintain health. The following trial investigated whether the stocking density affected gene expressions in immune organs.2. Jinding ducklings were raised in either low or high density (4 or 8 birds/m2) conditions from four to 14 weeks of age, and were then slaughtered and tissues removed. Samples were subjected to high-throughput sequencing to sequence RNA extraction. After filtering calculations with R software, a total of 508 (thymus) and 1,356 (bursa of Fabricius) differentially expressed genes (DEGs) were identified, suggesting that stocking density has an effect on gene expression in duck immune organs.3. Out of a total of 112 immune factor genes and 112 immune pattern receptor genes in ducks, four thymus and 18 bursa of Fabricius genes were differentially expressed in ducks, which indicated that the change of stocking density could affect the expression of immune genes in poultry.

Decreased expression of SorCS1 in colorectal cancer: An independent predictor of poor prognosis.

Previously, we identified that sortilin related VPS10 domain containing receptor 1 (SorCS1) was hypermethylated in colorectal cancer (CRC) tissues. Here, we aimed to investigate the association between CRC and SorCS1. DNA methylation was determined by methylation-specific polymerase chain reaction (MSP) or quantitative real-time methylation analysis (MethyLight). Colorectal cancer tissue specimens from 239 patients that had undergone surgical treatment were evaluated using immunohistochemistry (IHC) analysis for the expression of SorCS1 and correlated with clinicopathological variables and prognosis. We found that SorCS1 was hypermethylated in CRC cell lines and 67.5% (27/40) CRC tumor tissues. The loss of SorCS1 mRNA (p<0.001) and protein expression (p=0.033) were highly correlated with promoter methylation. In addition, SorCS1 expression was significantly increased in younger patients (p=0.006), low CEA level (p<0.001) and pT1-2 stage (p=0.005). Survival analysis revealed that decreased expression of SorCS1 was an independent factor for predicting the increased risk of recurrence (p=0.024) and poor overall survival (p=0.006). Subgroup analysis for CEA level, pT and pN classifications showed that SorCS1 retained its stratified significance only in patients with low CEA level, pT3-4 tumors and pN1-2 lymph node status. Our findings suggest that SorCS1 is epigenetically inactivated in a substantial fraction of CRC, and its expression may be a promising prognostic factor in CRC patients.

Temperature-sensitive pathways may be involved in duck embryonic developmental recovery from blastoderm dormancy during hatching.

1. Birds' newly oviposited blastoderms can survive several weeks in a dormant state during low-temperature storage. Previous studies demonstrated that there is a critical temperature range from 19 to 27°C for chicken embryos. Within this range, the embryo will diapause in a dormant state; once the temperature rises above this range, the blastoderm will break dormancy. 2. Clarifying the mechanism that initiates duck embryo developmental recovery from blastoderm dormancy will be helpful to change temperature control to improve hatching in poultry production. It was hypothesised that there might be some temperature-sensitive genes involved in initiating duck embryo developmental recovery from blastoderm dormancy. 3. To test this hypothesis, the transcriptome of the newly oviposited duck blastoderm and duck embryo (incubated for 48 hours) were sequenced to screen for differentially expressed genes with functions that had been predicted by bioinformatics. 4. The results showed that there were 2416 differentially expressed genes between the two groups, 53 of which were involved in temperature-sensitive pathways. The protein-protein interaction network combined these 53 temperature-sensitive genes and another group of 65 genes, which enriched the development pathway. These results suggested that temperature-sensitive genes may be involved in growth and development related pathways.

USING GUESSING GAME TO INCREASE COMPLICATION AWARENESS OF PATIENTS WITH NEWLY DIAGNOSED TYPE 2 DIABETES.

BACKGROUND: Prevention of complications is widely considered as the main aim of diabetes control. And diabetes education is the cornerstone for type 2 diabetes (T2D) management. However, traditional lecture-based diabetes patient education activities have small and short-lasting efficacy. Therefore, technology-based initiatives for diabetes patient education are urgently required. OBJECTIVE: To evaluate Guessing, a popular game, as tool in increasing complication awareness of patients with newly diagnosed T2D during diabetes care. PATIENTS AND METHODS: In a cohort study, 103 patients were split into Guessing Game group and control group. The opinions of patients and educators in Guessing Game group were surveyed. Patient performance was evaluated by test scores and the attendance to diabetes complication screening clinic. RESULTS: A majority of patients and all educators believed that Guessing Game enhanced complication awareness. Educatees achieved higher total scores and test scores in "Fill in the Gaps" (one of 2 types of test item), more actively attended complication screening clinic, after using Guessing Game as an education tool. CONCLUSION: Guessing Game is an attractive and effective educational intervention to increase complication awareness of T2D patients.

[Establishment of PCR assays and genetic polymorphism analysis of genes encoding Clostridium perfringens ß2 toxin from different sources].

Objective: To establish and optimize PCR methods for the gene encoding of Clostridium perfringens ß2 toxin (cpb2) and atypical-cpb2 (aty-cpb2), analyze the epidemiological characteristics and genetic polymorphism of the cpb2 of Clostridium perfringens in 9 Chinese areas from 2016 to 2021. Methods: The cpb2 of 188 Clostridium perfringens strains were examined by PCR; the cpb2 sequences were acquired by whole-genome sequencing to analyze the genetic polymorphism. Using Mega 11 and the Makeblastdb tool, a phylogenetic tree, and cpb2-library based on 110 strains carrying the cpb2 were produced. Using the Blastn technique, a comparison was made to discover sequence similarity between consensus-cpb2 (con-cpb2) and aty-cpb2. Results: The specificity of PCR assay for the cpb2 and aty-cpb2 was verified. The PCR results for cpb2 amplification were highly consistent with the whole-genome sequencing approach (Kappa=0.946, P<0.001). A total of 107 strains from nine regions in China carried cpb2, 94 types A strains carried aty-cpb2, 6 types A strains carried con-cpb2, and 7 types F strains carried aty-cpb2. The nucleotide sequence similarity between the two coding genes was 68.97%-70.97%, and the similarity between the same coding genes was 98.00%-100.00%. Conclusions: In this study, a specific PCR method for cpb2 toxin was developed, and the previous PCR method for detecting aty-cpb2 was improved. aty-cpb2 is the primary gene encoding of ß2 toxin. There is a significant nucleotide sequence variance between the various cpb2 genotypes.

[Study on the features of Clostridioides difficile infection among diarrhea patients in Kunming from 2018 to 2020].

Objective: We analyze the characteristics of Clostridioides difficile (C. difficile) infection among diarrhea patients in Kunming from 2018 to 2020 and provide evidence for follow-up surveillance and prevention. Methods: A total of 388 fecal samples of diarrhea patients from four sentinel hospitals in Yunnan Province from 2018 to 2020 were collected. Real-time quantitative PCR was used to detect the fecal toxin genes of C. difficile. The positive fecal samples isolated the bacteria, and isolates were identified by mass spectrometry. The genomic DNA of the strains was extracted for multi-locus sequence typing (MLST). The fecal toxin, strain isolation, and clinical patient characteristics, including co-infection with other pathogens, were analyzed. Results: Among the 388 fecal samples, 47 samples with positive reference genes of C. difficile were positive, with a total positive rate of 12.11%. There were 4 (8.51%) non-toxigenic and 43 (91.49%) toxigenic ones. A total of 18 strains C. difficile were isolated from 47 positive specimens, and the isolation rate of positive specimens was 38.30%. Among them, 14 strains were positive for tcdA, tcdB, tcdC, tcdR, and tcdE. All 18 strains of C. difficile were negative for binary toxins. The MLST results showed 10 sequence types (ST), including 5 strains of ST37, accounting for 27.78%; 2 strains of ST129, ST3, ST54, and ST2, respectively; and 1 strain of ST35, ST532, ST48, ST27, and ST39, respectively. Fecal toxin gene positive (tcdB+) results were statistically associated with the patient's age group and with or without fever before the visit; positive isolates were only statistically associated with the patient's age group. In addition, some C. difficile patients have co-infection with other diarrhea-related viruses. Conclusions: The infection of C. difficile in diarrhea patients in Kunming is mostly toxigenic strains, and the high diversity of strains was identified using the MLST method. Therefore, the surveillance and prevention of C. difficile should be strengthened.

[Progress in research of Clostridium perfringens toxin].

Clostridium perfringens can produce many kinds of toxins and hydrolase, causing gas gangrene, enteritis and enterotoxemia in both human and animals. It is known that C. perfringens can produce more than 20 toxins and hydrolases. The different toxin types are associated with specific disease types. At present, molecular toxin-typing method by PCR has replaced the traditional serological typing method. In this study, we systematically summarize the types, basic characteristics, pathogenic mechanism and the relationship with disease of C. perfringens toxins to provide evidence for the establishment of rapid detection method, immune antigen screening, antibody preparation and research of related pathogenic mechanism.