[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.

[Change of innate lymphoid cell subsets in peripheral bloods and ascites in liver cirrhotic patients complicated with spontaneous bacterial peritonitis].

Objective: To investigate the change of innate lymphoid cells (ILC) subsets in peripheral blood and ascites in liver cirrhotic patients complicated with spontaneous bacterial peritonitis (SBP). Methods: The data of 62 patients with liver cirrhosis admited to the Zhumadian Central Hospital from November 2019 to November 2020 were analyzed. Among them, 41 cases were complicated with untainted ascites (untainted ascites group), while the other 21 cases were complicated with SBP (SBP group). Meanwhile, 20 cases of controls who received healthy examination in the same period were also enrolled (control group). Peripheral blood mononuclear cell (PBMC) was isolated from peripheral blood of all patients and controls. Mononuclear cell in ascites was isolated from patients with liver cirrhosis. The percentage of ILC1, ILC2, and ILC3 subsets in PBMC and mononuclear cell in ascites were measured by flow cytometry. CD3-CD19-CD20-CD14- cells (lin-cells) were purified from ascites and were stimulated with lipopolysaccharide (LPS) for 24 h. The transcription factor T-bet, GATA3, and RORγt mRNA relative level in lin-cells was semi-quantified by real-time PCR. Cytokine level in the supernatants was measured by enzyme linked immunosorbent assay. Differences of ILC subsets in peripheral blood and ascites were compared among groups. Results: There were twenty-nine males and twelve females in untainted ascites group, aged M(Q1,Q3) 49(33, 78) years. There were twelve males and nine females in SBP group, aged 50(37, 76) years. There were eleven males and nine females in control group, aged 48(32, 69) years. lin-CD45+CD161+CD127+ ILC cells could be detected in both peripheral blood and ascites. There was no significant difference in total ILC percentage within PBMC among untainted ascites group, SBP group, and control group (P=0.235). There was also no significant difference of total ILC percentage within mononuclear cells in ascites between untainted ascites group and SBP group (P=0.232). The differences were not statistically significant of peripheral CD117-CRTh2-ILC1, CRTh2+ILC2, or CD117+CRTh2-ILC3 within peripheral ILC among untainted ascites group, SBP group, and control group (all P>0.05). ILC1 percentage in ascites was up-regulated in SBP group compared with untainted ascites group (35.69%±3.39% vs 26.40%±3.85%, P<0.001), while ILC2 in ascites was down-regulated in SBP group (36.83%±7.70% vs 48.35%±9.45%, P<0.001). There was no statistical difference in ILC3 percentage in ascites between the two groups (P=0.230). T-bet mRNA relative level and IFN-γ production by lin- cells from ascites were elevated in response to LPS stimulation in SBP group compared with untainted ascites group (both P<0.001). GATA3 mRNA relative level and IL-5/IL-13 secretion by lin-cells from ascites were reduced in SBP group compared with untainted ascites group (both P<0.05). There was no significant difference of RORγt mRNA relative level or IL-17/IL-22 expression between the two groups (both P>0.05). Conclusion: Peripheral ILC subsets do not change in liver cirrhosis patients with SBP. ILC1 percentage is up-regulated, and ILC2 percentage is down-regulated in ascites in liver cirrhosis patients with SBP.

[Roles of detection, surveillance and early warning on outbreaks or epidemics of infectious diseases].

Infectious diseases are still one of the leading causes of morbidity and death globally, affecting public health and life, social and economic development, and even national security. Early detection focuses on detecting the abnormal information of infectious disease outbreaks or epidemics in a timely and sensitive way to conduct field investigation and verification. It is also a precursor to effective surveillance and early warning system. The effective surveillance and early warning system can fully and accurately understand the real conditions, driving forces, and transmission chain of the occurrence of a specific infectious disease outbreak and epidemic and put forward scientific and effective prevention and control strategies and measures. Due to the measurement of the resources support and the particular data collection value, it is not easy to obtain epidemiological, etiological, and other data information in a timely, complete and accurate manner. This paper summarized the theory and technology on early detection, effective surveillance, and early warning information on infectious diseases. It also integrated and utilized the multi-source data, including effective infectious disease surveillance and the country's early warning system, to better understand the outbreak epidemic, causes, risks, processes, and driving forces. Thus, it is possible to set up a sensitive, specific staging measurement innovative technical system to monitor, early warning, and timely respond to acute infectious diseases through multidisciplinary cooperation in China. It provides the basis for strengthening the surveillance and early warning of new emerging and major infectious diseases and public health emergencies, avoiding the spread of inadequate response to infectious disease, and preventing the resources waste of over-response.

[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.

Long non-coding RNA OR3A4 promotes metastasis of ovarian cancer via inhibiting KLF6.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Long non-coding RNA OR3A4 promotes metastasis of ovarian cancer via inhibiting KLF6, by F.-F. Guo, M.-M. Jiang, L.-L. Hong, B. Qiao, X.-M. Lin, W.-Y. Xu, X.-Q. Fu, published in Eur Rev Med Pharmacol Sci 2019; 23 (6): 2360-2365-DOI: 10.26355/eurrev_201903_17380-PMID: 30964160" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/17380.

Long non-coding RNA OR3A4 promotes metastasis of ovarian cancer via inhibiting KLF6.

OBJECTIVE: Recently, long noncoding RNAs (lncRNAs) have attracted much attention for their roles in tumor progression. The aim of this study was to investigate the exact role of lncRNA OR3A4 in the development of ovarian cancer (OC), and to explore the possible underlying mechanism. PATIENTS AND METHODS: OR3A4 expression in OC tissue samples was detected by Real-time quantitative polymerase chain reaction (qRT-PCR). Moreover, wound healing assay and transwell assay were performed to evaluate the effect of OR3A4 on the metastasis of OC. Furthermore, the underlying mechanism was explored by RT-qPCR and Western blot assay. RESULTS: The expression level of OR3A4 in OC samples was significantly higher than that of adjacent tissues. Moreover, cell migration and invasion were significantly repressed after OR3A4 knockdown in vitro. Moreover, the mRNA and protein expressions of kruppel-like factor 6 (KLF6) were remarkably upregulated after knockdown of OR3A4. Furthermore, the expression level of KLF6 was negatively correlated with the expression of OR3A4 in OC tissues. CONCLUSIONS: Our results showed that OR3A4 could enhance OC cell metastasis and invasion via suppressing KLF6. Moreover, OR3A4 might be a potential therapeutic target for OC.

Research on the expression of MRNA-518b in the pathogenesis of placenta accreta.

OBJECTIVE: To investigate the expression of micro ribonucleic acid miR-518b and its regulatory role in the pathogenesis of placenta accreta. PATIENTS AND METHODS: A total of 50 parturient women in the Obstetric Department were collected and divided into observation group (placenta accreta, n=23) and control group (normal placenta, n=27). After the placental tissues were removed via surgery, the expressions of osteopontin (OPN) and vascular endothelial growth factor (VEGF) were detected using the immunohistochemical method. The relative expression levels of OPN and VEGF proteins were detected via Western blotting, and the relative expression levels of OPN messenger RNA (mRNA), VEGF mRNA and miR-518b were detected via quantitative polymerase chain reaction (qPCR). Moreover, the correlations of miR-518b with OPN mRNA and VEGF mRNA were studied via Pearson correlation analysis. RESULTS: Compared with those in control group, the expressions of OPN and VEGF proteins in observation group were significantly increased, while the levels of OPN mRNA, VEGF mRNA and miR-518b in observation group were also significantly elevated (p<0.05). There were positive correlations between miR-518b and levels of OPN, VEGF mRNA. CONCLUSIONS: The high expression of miR-518b may lead to the development of placenta accreta through upregulating the transcription and protein expression of downstream VEGF and OPN, providing insights for the future therapy against the pathogenesis of placenta accreta.

Molecular identification of variety purity in a cotton hybrid with unknown parentage using DNA-SSR markers.

Molecular identification of hybrid purity is difficult in regional trials of cotton varieties and hybrid trials. In particular, the molecular detection of hybrid purity has not yet been reported in the case of unknown parentage. In this study, we screened 5000 pairs of primers and chose 17 pairs of core simple sequence repeat (SSR) primers to determine the F1 purity of Han6402. The results showed that the purity based on SSR markers reached 100%. Twelve of the 17 pairs of primers exhibited co-dominant banding patterns, and 5 showed non-co-dominant banding patterns. Moreover, we constructed an F1 SSR fingerprinting profile that enabled the identification of the authenticity of Han 6402. Using these primers, we subsequently detected 44 individual F2 seedlings, and the results exhibited different extents of separation, in which the majority of genotypes were heterozygous with co-dominance at most of the loci that differed from each other. The results validated the underlying heterozygous status of the F2 population at the molecular level. Therefore, we conclude that the set of core SSR primers can be used for the laboratory identification of the authenticity and purity of cotton hybrids, not only for distinguishing Fl hybrids or segregating F2 populations, but also for detecting volunteer seeds as fake F1 hybrids in the cotton hybrid industry, based on the hybrid fingerprinting.

[Clinical stages and outcomes of severe cases on hand, foot and mouth disease].

Objective: To understand the characteristics and relation of clinical stage and outcome of severe cases on hand, foot and mouth disease (HFMD) and to establish the evaluation method for understanding severity of this disease. Methods: According to factors as geographical location, economic and epidemic levels, five provinces (Henan, Shandong, Yunnan, Zhejiang and Sichuan provinces) were selected. Reported severe cases of HFMD from the National Notifiable Diseases Reporting System were selected randomly in the five provinces. Basic epidemiological information, clinical data, and pathogen testing results in the involved hospitals were collected. Clinical stages on all the patients were decided in accordance with"the clinical expert consensus on diagnosis and treatment for severe case of enterovirus type 71 (EV71) infections (2011 edition)" . Data were analyzed using SPSS software 18.0 and other epidemiological methods. Results: A total of657 severe HFMD cases were investigated, with 326 cases positive of EV71, accounting for 91.3% (326/357) among all the laboratory-confirmed cases. Of the 657 cases, 542 cases (82.5%, 95%CI: 79.4%-85.3%) were diagnosed as in stage 2 (with nervous system involvement), 99 cases (15.1%, 95%CI: 12.4%-18.0%) in stage 3 (early phase of function failure on heart and lung), and 16 cases (2.4%, 95%CI: 1.4%-3.9%) were in stage 4 (function failure of heart and lung). 11 cases (1.7%, 95%CI: 0.9%-3.0%) were with squeal when discharged from hospital with 8 cases (1.2%, 95%CI: 0.6%-2.3%) died. When comparing the proportions among stage 2, stage 3 and stage 4, significant differences were found between age groups (χ(2)=22.632, P=0.012). The younger the patient was the lower the proportions of stage 2 and the more proportion of stage 3 appeared. When comparing the proportions of clinical stages among the five provinces, significant differences (χ(2)=41.481, P=0.000) were noticed. Proportions of different clinical stages in gender, ethnicity, occupation, place of residence types and the type of pathogen appeared no significant differences, respectively. However, the proportions of squeal and death in stage 2, stage 3 and stage 4 showed significant differences (sequela: χ(2)=12.960, P=0.001; Death: χ(2)=16.850, P=0.001), respectively. Conclusions: The percentage of clinical stages of severe HFMD patients related to the rate of squeal and death. Clinical staging can be used for assessing the clinical severity of complications and the effectiveness of treatment, of HFMD.

Cloning and characterization of DELLA genes in Artemisia annua.

Gibberellins (GA) are some of the most important phytohormones involved in plant development. DELLA proteins are negative regulators of GA signaling in many plants. In this study, the full-length cDNA sequences of three DELLA genes were cloned from Artemisia annua. Phylogenetic analysis revealed that AaDELLA1 and AaDELLA2 were located in the same cluster, but AaDELLA3 was not. Subcellular localization analysis suggested that AaDELLAs can be targeted to the nucleus and/or cytoplasm. Real-time PCR indicated that all three AaDELLA genes exhibited the highest expression in seeds. Expression of all AaDELLA genes was enhanced by exogenous MeJA treatment but inhibited by GA3 treatment. Yeast two-hybrid assay showed that AaDELLAs could interact with basic helix-loop-helix transcription factor AaMYC2, suggesting that GA and JA signaling may be involved in cross-talk via DELLA and MYC2 interaction in A. annua.

Cloning and characterization of the drought-resistance OsRCI2-5 gene in rice (Oryza sativa L.).

The genomic expression profile of the super-hybrid rice Liangyoupeijiu female parent Pei'ai 64S in different tissues at different developmental stages under low temperature, drought, and high temperature stresses were detected using an Affymetrix GeneChip Rice Genome Array to screen upregulated and downregulated genes. In this study, we screened the drought-resistant gene OsRCI2-5, after which a constitutive OsRCI2-5 construct was created and transferred into Nipponbare. After polyethylene glycol-6000 and drought treatment, we found that the OsRCI2-5 gene improved the drought resistance of Nipponbare. Gene expression profiling showed that the OsRCI2-5 gene was expressed in the rice leaves, stems, and flower organs. Subcellular localization revealed that the gene was located in the membranes, and hence, we can deduce that a membrane signal peptide was responsible for signal transduction.

The AKT1/NF-kappaB/Notch1/PTEN axis has an important role in chemoresistance of gastric cancer cells.

The inherent resistance of tumors to DNA damage often limits the efficacy of chemotherapy. The aim of this work is to explore the potential mechanism for development of chemoresistance in gastric cancer. Our data revealed that AKT1 mRNA and protein expression were induced by doxorubicin (a chemotherapeutic agent); the doxorubicin-induced AKT1 expression and activation increased the binding of NF-kappaB on Notch1 DNA promoter and then promoted the Notch1 transcription and expression; enhanced expression of Notch1 further upregulated PTEN expression through CBF-1 binding to PTEN DNA promoter; and inhibition of AKT1 expression and activity sensitized the gastric cancer cell to doxorubicin treatment in cultured gastric cancer cell lines and xenograft nude mice gastric cancer model. Furthermore, our data demonstrated that both Notch1 and PTEN were absent or minimally expressed in gastric cancer tissue but abundant in paired normal gastric mucosa, and the expression of Notch1 correlated with that of PTEN. Together, these novel results suggested that a novel AKT1/NF-kappaB/Notch1/PTEN axis has an important role in the development of chemoresistance in gastric cancer. Notch1 has an anti-cancer role in gastric cancer.

Noise-like pulse in a gain-guided soliton fiber laser.

We report on the operation of a passively mode-locked fiber ring laser made of purely positive dispersion fibers and mode-locked by using the nonlinear polarization rotation technique. It was experimentally found that apart from the gain-guided soliton operation the laser can also emit a kind of noise-like pulse. We show numerically that the noise-like pulse emission is caused by the peak power clamping effect of the laser cavity on the gain-guided soliton.