New primers for sex identification in the Chukar partridges (Alectoris chukar).

1. Male and female Chukar partridges are difficult to differentiate based on their morphology or by the Chromobox-Helicase-DNA binding (CHD) during early growth.2. The current study developed a novel, simple, low-cost and rapid sexing protocol for Chukar partridges based on the newly defined sexing gene ubiquitin-associated protein 2 (UBAP2).3. The length of polymorphism between UBAP2-W and UBAP2-Z homologous genes allows for easy sex discrimination in this species. Molecular sexing analysis was based on the simultaneous amplification of both genes, resulting in two distinct amplicons (947 bp and 535 bp) in heterogametic females and only a single band (535 bp) in homogametic males, which is easy to detect with agarose gel electrophoresis.4. This technique is simple and convenient for genetic sex determination in Chukar partridges.

A new method for molecular sex identification in the emu (Dromaius novaehollandiae).

1. Sex chromosomes of emus are largely homomorphic. Therefore, the standard methodology for molecular sexing based on screening intron length variations in sex-linked genes is not applicable. However, emu sexing requires costly and time-consuming PCR-RFLP or multiplex PCR methods.2. This experiment used a directed PCR amplification and capillary electrophoresis sexing protocol. Two distinct peaks were observed in females (ZW), while only one peak was observed in males (ZZ).3. This sexing technique proved to be rapid, non-invasive, and highly sensitive and may be useful for verifying the sex ratio and breeding management of emus.

Effects of milk powder intervention on bone mineral density and indicators related to bone metabolism in Chinese adolescents.

The study is to determine the effects of milk powder on bone density and metabolism in healthy adolescents. Vitamin D and calcium supplements increased IGF-1 but did not affect bone mineralization or turnover. Higher vitamin D in combination with sufficient calcium supplementation in such populations requires attention. INTRODUCTION: Both calcium and vitamin D play an important role in bone mineralization in adolescents. METHODS: In this one and a half-year randomized controlled trial, 232 participants (aged 12-15 years) were randomly assigned to three intervention groups receiving milk powder fortified with vitamin D 400 IU plus calcium 300, or 600, or 900 mg [Ca3D (n = 54), Ca6D (n = 56), and Ca9D (n = 49)], or one control group maintaining habitual diet [control (n = 73)]. Bone turnover markers, serum intact PTH, 25(OH)D, and IGF-1 levels were measured at baseline and one and a half years, and bone mineral contents and bone areal mineral density were measured by Dual-energy X-ray absorptiometry (DXA) at baseline, one year, and one and a half years. RESULTS: Baseline average serum 25(OH)D level and calcium intake were 29.4 nmol/L and 363.5 mg/day, respectively. There was a significant increase in bone turnover, total body, hip, lumbar spine bone mineral density (BMD), and total body BMC as well as slight fluctuations in 25(OH)D levels over one and a half years without between-group differences. Significantly decreased serum PTH level was only observed in the Ca6D group (31%, p < 0.0001), in which the intervention effect was also significant (p = 0.0029) compared with the control group. IGF-1 levels increased significantly in all intervention groups (18.5 to 22.8%, p < 0.05) but decreased in the control group (16.5%, p < 0.05), and the group by time interaction was also significant (p = 0.0029). CONCLUSIONS: Our study showed that in healthy adolescents with low vitamin D status and calcium intake, mild vitamin D and mild to modest calcium supplements increased IGF-1 but did not affect bone mineralization or turnover. Higher vitamin D in combination with sufficient calcium supplementation in such populations requires attention.

Genetic diversity of Jiangsu native chicken breeds assessed with the mitochondrial DNA D-loop region.

1. The objective of this study was to determine the origin and evolution of chickens from 5 native breeds that are traditionally raised in Jiangsu Province. 2. To address this question, the complete mitochondrial DNA D-loop sequence of 149 chickens from 5 native breeds of Jiangsu Province was analysed. 3. Sequence read lengths of the native breeds were 1231 to 1232 bp, with a single-base deletion from the 859 bp site in the 1231 bp haplotype. A total of 33 variable sites that defined 19 haplotypes were identified. The average haplotype diversity and nucleotide diversity were 0.862 ± 0.017 and 0.00591 ± 0.00135. 4. Phylogenetic analysis showed that genetic structure of the mtDNA haplotypes of Jiangsu chickens are distributed across 5 clades (haplogroups): Clades A, B, C, D, and E. However, most of the individuals characterised in this study belonged to clades A and B. 5. The results of this study indicate that Jiangsu chicken populations have relatively low nucleotide and haplotype diversity and likely share 5 common maternal lineages.

[The relationship between non-alcoholic fatty liver disease and metabolic syndrome in patients with latent autoimmune diabetes in adults].

Objective: To investigate the relationship between non-alcoholic fatty liver disease (NAFLD) and metabolic syndrome (MetS) in patients with latent autoimmune diabetes in adults (LADA). Methods: A total of 223 hospitalized patients were recruited between January 2007 and June 2009 in Department of Endocrinology and Metabolism of the Sixth People's Hospital Affiliated to Shanghai Jiaotong University. Finally, 142 patients with complete clinical data and without history of drinking were included in this study. According to the Chinese Medical Association's Guidelines of NAFLD, based on the result of ultrasound, all subjects were divided into two groups including patients with LADA and NAFLD (n=37) and patients with LADA but without NAFLD (n=105). Clinical data including diabetes duration, history of smoking and medications, height, weight, blood pressure, blood lipids, blood glucose, C-peptide, and liver and kidney function were collected. The prevalence and components of MetS were compared between two groups. The association between MetS and NAFLD was also explored. Results: After adjusting for age and sex, compared with the subjects without NAFLD, the subjects with NAFLD were older and had higher percentage of hypertension, and had higher body mass index[(26.5±3.7) kg/m(2) vs (21.9±3.1) kg/m(2)], waist-hip ratio(0.92±0.06 vs 0.86±0.07), low density lipoprotein cholesterol[(3.26±0.72) mmol/L vs (2.70±0.87) mmol/L], C-reactive protein, fasting C-peptide, 2 h postprandial C-peptide, systolic blood pressure, diastolic blood pressure, alanine aminotransferase and triglyceride (all P<0.05). But they had lower high-density lipoprotein cholesterol[(1.17±0.43) mmol/L vs (1.35±0.40) mmol/L]and HbA1c[(8.83±2.14) % vs (10.02±2.79)%](both P<0.05). In addition, after adjusting for age and sex, compared with the patients with LADA but without NAFLD, the prevalence of MetS in the patients with LADA and NAFLD was obviously higher (97.3% vs 47.6%, P<0.001), and the proportion of the patients with four (32.4% vs 16.2%, P<0.001) and five (43.2% vs 5.7%, P<0.001) components of MetS in the patients with LADA and NAFLD was also significantly increased than that in the patients with LADA but without NAFLD. Binary regression analysis showed that NAFLD was an independent factor associated with MetS in the patients with LADA after correcting other confounding factors (P<0.001). Conclusions: Compared with the patients with LADA but without NAFLD, the prevalence of MetS was obviously higher, and had more serious metabolic disorder in the patients with LADA and NAFLD. The presence of NAFLD was an independent factor associated with MetS in the patients with LADA.

Protein and mRNA expression of follicle-stimulating hormone receptor and luteinizing hormone receptor during the oestrus in the yak (Bos grunniens).

Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) have a central role in follicle growth, maturation and oestrus, but no clear pathway in the seasonal oestrus of yak (Bos grunniens) has been found. To better understand the role of FSH and LH in seasonal oestrus in the yak, six yaks were slaughtered while in oestrus, and the pineal gland, hypothalamus, pituitary gland, and gonads were collected. Using real-time PCR and immunohistochemical assays, we determined the mRNA and protein expression of the FSH and LH receptors (FSHR and LHR) in these organs. The analysis showed that the FSHR mRNA expression level was higher in the pituitary gland tissue compared with LHR (p < .01) during oestrus. By contrast, there was low expression of FSHR and LHR mRNA in the pineal gland and hypothalamus. FSHR mRNA expression was higher than that of LHR (p < .05) in the ovary, whereas LHR mRNA expression was higher than that of FSHR (p < .01) in the uterus. FSHR and LHR proteins were located in the pinealocyte, synaptic ribbon and synaptic spherules of the pineal gland and that FSH and LH interact via nerve fibres. In the hypothalamus, FSHR and LHR proteins were located in the magnocellular neurons and parvocellular neurons. FSHR and LHR proteins were localized in acidophilic cells and basophilic cells in the pituitary gland, and in surface epithelium, stromal cell and gland epithelium in the uterus. In the ovary, FSHR and LHR protein were present in the ovarian follicle. Thus, we concluded that FSHR and LHR are located in the pineal gland, hypothalamus, pituitary and gonad during oestrus in the yak. However, FSHR was mainly expressed in the pituitary gland and ovaries, whereas LHR was mainly expressed in the pituitary gland and uterus.

[The vitamin D nutritional status in Chinese urban women of child-bearing age from 2010 to 2012].

Objective: To evaluate the vitamin D nutritional status in Chinese women of child-bearing age by analyzing serum 25-hydroxyvitamin D level in 2010-2012. Methods: Data were obtained from the China Nutrition and Health Survey in 2010-2012. Using cluster sampling and proportional stratified random sampling, 1 514 women of child-bearing age (18-44 years old) from 34 metropolis and 41 small and medium-sized cities were included in this study. Demographic information was collected by questionnaire and serum 25-hydroxyvitamin D concentration was determined by radioimmunoassay, in accordance with the 2010 Institute of Medicine of the National Academies standards. We compared differences in vitamin D levels, specifically serious deficiency, lack of deficiency, insufficiency, and excess. Results: The overall serum 25-hydroxyvitamin D level of Chinese urban women of child-bearing age (P(50) (P(25)-P(75))) was 20.1 (15.1-26.3) ng/ml; minorities had a significantly higher serum 25-hydroxyvitamin D level of 22.0 (15.9-27.5) ng/ml compared with women of Han nationality (19.8 (14.9-26.2) ng/ml) (χ(2)=7.02, P=0.008). The proportions of women with serious deficiency, lack of deficiency, insufficiency, and excess vitamin D were 11.6% (n=175), 37.9% (n=574), 35.1% (n=531), and 0.3% (n=5), respectively. Only 15.1% (n=229) of women of child-bearing age had normal vitamin D nutritional status. No significant differences in vitamin D nutritional status were observed according to age, body mass index, city, nationality, educational level, marital status, or household income per capita (P>0.05). Conclusion: Most Chinese urban women of child-bearing age have poor vitamin D levels and require vitamin D supplementation.

[Progress in Application of Measuring Skeleton by CT in Forensic Anthropology Research].

Individual identification by measuring the human skeleton is an important research in the field of forensic anthropology. Computed tomography （CT） technology can provide high-resolution image of skeleton. Skeleton image can be reformed by software in the post-processing workstation. Different skeleton measurement indexes of anthropology, such as diameter, angle, area and volume, can be measured on section and reformative images. Measurement process is barely affected by human factors. This paper reviews the literatures at home and abroad about the application of measuring skeleton by CT in forensic anthropology research for individual identification in four aspects, including sex determination, height infer, facial soft tissue thickness measurement and age estimation. The major technology and the application of CT in forensic anthropology research are compared and discussed, respectively.

Characterization of the complete mitochondrial genome and phylogenetic relationships of the three-spot swimming crab (Portunus sanguinolentus).

In this study, we determined the whole mitochondrial genome profile of the three-spot swimming crab (Portunus sanguinolentus) and elucidated phylogenetic relationships between representative species in the order Decapoda. The mitochondrial genome was 16,024 bp in length and consisted of 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a putative control region. Of the 37 genes, 23 were encoded by the heavy strand while 14 were encoded by the light strand. Four types of start codons were identified; ATG initiated nine genes, ATT initiated two genes, and ATC and GTG each started one gene. Nine protein-coding genes ended with a complete TAA or TAG stop codon, and four genes ended with an incomplete T or TA codon. Fourteen non-coding regions were found, which ranged from 1 to 34 bp in length. Nine overlaps were observed, with lengths between 1 and 7 bp. Phylogenetic analysis suggested that P. sanguinolentus is genetically closest to P. trituberculatus and P. pelagicus. Charybdis feriata, C. japonica, and Thalamita crenata formed a single cluster, and were close to the genera Callinectes and Portunus. Therefore, the genera Charybdis and Thalamita should be classified into the subfamily Portuninae.

Characterization of the complete mitochondrial genome of Portunus pelagicus with implications for phylogenomics.

This study determined the mitochondrial genome structure of the blue swimming crab (Portunus pelagicus), and elucidated its phylogenetic relationships among the species within the order Decapoda. The complete mitochondrial genome was 16,155 bp long, and contained 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and 1 DNA control region. The gene order of the genome was the same as that found within the family Portunidae. Twenty-three genes were on the heavy strand and 14 were on the light strand. Almost all of the protein-coding genes were initiated by an ATG codon, except for three genes (ATP6, ND1, and ND3) that started with a rare ATT codon. Of the 13 protein-coding genes, 10 ended with complete TAA or TAG stop codons and three ended with an incomplete T codon. Thirteen non-coding regions were identified that ranged from 1 to 30 bp in length. Nine overlaps were found, which ranged 1 to 7 bp in length. Phylogenetic analyses based on 12 concatenated protein-coding genes revealed that P. pelagicus formed a monophyletic group with Portunus trituberculatus, which were in a larger group with Callinectes sapidus, while the genera Charybdis and Thalamita formed another group. These two groups clustered together and grouped with the genus Scylla. The phylogenetic analysis supported the inclusion of Charybdis in subfamily Portuninae of the family Portunidae, and revealed a close relationship between Charybdis and Thalamita. We suggest that Thalamita should also be classified into the subfamily Portuninae. The results can be used in the study of phylogenetic, population genetic and conservation genetics of P. pelagicus.

Sarcopenia, as defined by low muscle mass, strength and physical performance, predicts complications after surgery for colorectal cancer.

AIM: Recent studies have shown that sarcopenia is associated with negative postoperative outcomes. However, none of these studies analysed muscle strength or physical performance, which are also important components of sarcopenia. The present study aimed to investigate whether sarcopenia itself, as defined by low muscle mass, strength and physical performance, would predict complications after surgery for colorectal cancer. METHOD: We conducted a prospective study of patients who underwent surgery for colorectal cancer at our department between August 2014 and February 2015. Sarcopenia was diagnosed by a combination of third lumbar vertebra muscle index (L3 MI), handgrip strength and 6-m usual gait speed. Univariate and multivariate analyses evaluating the risk factors for postoperative complications were performed. Only complications classified as Grade II or above according to the Clavien-Dindo classification were analysed in this study. RESULTS: A total of 142 patients were included in the study, and 17 patients were diagnosed as having sarcopenia. Postoperative complications of Grade II or above occurred in 40 patients, including 10 with sarcopenia and 30 without sarcopenia. Multivariate analysis showed that sarcopenia and previous abdominal surgery were independent risk factors for postoperative complications. Patients with sarcopenia also had an obvious tendency to a higher incidence of infectious complications. By comparing two logistic regression models, sarcopenia showed a better predictive power for postoperative complications than did low muscle mass. CONCLUSION: Sarcopenia and previous abdominal surgery are independent risk factors for complications after surgery for colorectal cancer. Including a functional aspect to the definition of sarcopenia may result in a better prediction of postoperative complications.

Loss of STAG2 causes aneuploidy in normal human bladder cells.

The aim of this study was to determine how the function of human stromal antigen 2 (STAG2) plays an important role in proper chromosome separation. STAG2 mRNA in normal bladder cells and bladder tumor cells was evaluated by RT-PCR. The protein levels of STAG2 in normal bladder cells and bladder tumor cells were determined by western blot. A cell proliferation assay was used to measure the growth of tumor cells and STAG2-inhibited normal cells, and STAG2- inhibited normal cells were subjected to karyotype analysis. Both STAG-2 mRNA and protein expression levels were lower in bladder cancer cells compared to the controls. Knockdown of STAG2 caused aneuploidy in normal bladder cells, leading to a decreased expression of the cohesin complex components SMC1, SMC3 and RAD21, but there was no obvious effect of STAG2 knockdown on cell proliferation. Our study indicated that abnormal expression of STAG2 could cause aneuploidy in normal bladder cells.

Mapping codon usage in sequence regions flanking cleavage positions in the hepatitis A virus polyprotein.

To analyze the synonymous codon usage patterns of sequence regions flanking cleavage sites in the hepatitis A virus (HAV) polyprotein, the codon usage bias at codon positions and the synonymous codon usage in the target contexts of 30 virus strains were estimated by two simple methods that were based on the values for relative synonymous codon usage. In addition, the pattern of synonymous codon usage was compared between the genomic sequences in HAV and those of its human host. Our results indicated that HAV adopts a combination of coincidence and antagonism with the synonymous codon usage in humans. This characteristic may help HAV to efficiently use the translational machinery in its human host. We also observed that codon usage exhibited a strong bias in some specific positions in these contexts, and that the underrepresented synonymous codons, CUA for Leu, ACG for Thr, GUA for Val, and UCG for Ser, are preferentially used in these positions. These underrepresented synonymous codons likely play roles in regulating the rate of protein translation and influencing the secondary structure of the sequence regions flanking the cleavage sites.

[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.

[Exploration on the construction of analysis indicators system for antibiotic resistance monitoring].

Antibiotic resistance (AR) is a severe and fast-growing public health challenge with rapid globalization, especially in China. Although some monitoring systems were established in different fields, fragmentation of information failed to show the overall trend and spread of AR. It is necessary to establish a national monitoring system to reveal the occurrence, development, and spread of AR. The new AR monitoring system needs an updated analysis indicators system. We intend to recommend a new analysis indicators system for AR was constructed and applied to AR data monitoring and analysis for humans, animals, the environment, and foods. After investigating and analyzing the 5 Chinese major AR monitoring systems and literature, we have formulated 15 AR monitoring analysis indicators and initially established an evaluation system for the country's new AR monitoring system.

[Interpretation of group standard for Clostridioides difficile infection diagnosis].

Clostridioides difficile is a key pathogen of antibiotic related diarrhea and hospital associated infection, causing several outbreaks in Europe and North Americans and resulting in severe disease burden. However, the standardized diagnostic principle and detection specifications in C. difficile infection (CDI) survey are limited in China, and the infection rate and disease burden of CDI in China are unclear. Therefore, National Institute for Communicable Disease Control and Prevention,National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, together with another 11 institutions, draft the group standard entitled "Diagnosis of Clostridium difficile infection (T/CPMA 008-2020)" of Chinese Preventive Medicine Association. Based on the principle of "legality, scientificity, advancement, and feasibility", this standard clarifies risk factors, diagnosis principles, diagnoses and differential diagnoses in order to improve the accuracy of CDI diagnosis in clinical practice, guide the surveillance for CDI, and understand the infection rate and disease burden of CDI in China.

MiR-92b inhibits proliferation and invasion of lung cancer by targeting EZH2.

OBJECTIVE: To verify that miR-92b inhibits proliferation and invasion of lung cancer by targeting EZH2. MATERIALS AND METHODS: The expression levels of miR-92b and EZH2 in human bronchial epithelial cell line BEAS-2B and human lung cancer cell line (A549, NCI-H23, NCI-H358, NCI-H1975, PC-9) were detected, and miR-92b mimic, sh-EZH2 expression vector, and plasmid blank vector (blank group) were constructed. Blank group, miR-92b mimic, miR-92b mimic+sh-EZH2 group (combined group) were set up, MTT and transwell were used to detect the proliferation and invasion ability of A549 and NCI-H23 cells, and fluorescein report verified the regulatory relationship of miR-92b to EZH2. RESULTS: The expression level of miR-92b in A549, NCI-H23, NCI-H358, NCI-H1975, and PC-9 cells was lower than that in BEAS-2B cells (p<0.05). The expression level of EZH2 was higher than that of BEAS-2B cells (p<0.05). A549 and NCI-H23 cells were selected for transfection. After that, the expression level of miR-92 in miR-92b mimic, combined group A549 and NCI-H23 cells was higher than that in blank group (p<0.05), and miR-92b mimic had no difference with joint group (p>0.05). The expression level of EZH2 in cells of miR-92b mimic, blank group A549, and NCI-H23 was lower than that of combined group (p<0.05), and miR-92b mimic was lower than that of blank group (p<0.05). After the overexpression of miR-92b, pmirGLO-EZH2-3'UT Wt luciferase activity decreased significantly (p<0.05) but had no effect on pmirGLO-EZH2-3'UTR Mut Luciferase activity (p>0.05). Cell proliferation ability and invasion ability of A549 cells and NCI-H23 cells in miR-92b mimic group were lower than those in blank group (p<0.05), while those in combined group were higher than those in miR-92b mimic group (p<0.05). CONCLUSIONS: MiR-92b inhibits proliferation and invasion of lung cancer cells through targeted inhibition of EZH2, which is a potential target for future treatment of lung cancer.

[Standardization and application on ribotyping library of Clostridioides difficile in China].

Objective: To establish a standard operation procedure (SOP) for ribosome genotyping (ribotyping) on Clostridioides (C.) difficile, supplement and verify ribotyping typing library, so as to improve the comparability of data between different laboratories and to develop surveillance network of C. difficil in China. Methods: Molecular typing of 54 reference strains from the United States and Europe of C. difficile were performed by using the SOP referencing correspondence from abroad and from our laboratory with a BioNumerics 7.6 software to estimate the reference library of types of C. difficile. Identification of 374 clinical and animal isolates of C. difficile from 13 cities in China between 2010 and 2018, to supplement the library information. Kappa test was used to evaluate the consistency. Results: Results of capillary electrophoresis of reference strains appeared clear and stable, which guaranteed the clustering results being fast and accurate. Results from the supplementary typing showed that there were 84 types of isolates, of which 25 RT types were consistent with reference strains from abroad, while 58 RT types were different from referenced types. In the 40 referenced types, 15 RT types were not found in this study. In the consistency evaluation, the Kappa value was 0.891 and (P<0.01), showing the two Molecular typing as consistent and with close resemblance. Conclusions: The result of capillary electrophoresis by applying SOP for ribotyping on C. difficile base on QIAxcel capillary electrophoresis system, appeared clear and stable. The standardized library seemed more easily used for comparability and data sharing between the laboratories.