Phage therapy is one of the most important tools for the treatment of infections with multi-drug resistant bacteria. Such phages are usually isolated from hospital effluents, however, no systematic study on the distribution of phages in hospital effluents has been conducted so far. The aim of this study was to isolate the corresponding phages of common pathogenic bacteria isolated in the clinic as hosts, so as to assess the ecological distribution of phages in hospital wastewater and to provide a reference for the isolation and application of phages of drug-resistant bacteria in the clinic. A cross-sectional study design was used in this study. The 125 pathogenic bacteria (belonging to 16 different strains) isolated from the clinical microbiology laboratory of Qilu Hospital of Shandong University from May to June 2023 were selected as the target strains, and the phages corresponding to these strains were isolated and purified from the hospital wastewater by using the double-layer plate sandwich method. At the same time, the distribution of pathogenic bacteria in the same batch of wastewater was analyzed with the help of mNGS sequencing technology, so as to preliminarily investigate the abundance correspondence between pathogenic bacteria and phages in wastewater. The results showed that a total of 56 phage strains were isolated from 125 clinical pathogens as hosts, corresponding to six pathogens, including Acinetobacter baumannii, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. All six pathogenic bacteria contained strains with different degrees of drug resistance, with a higher percentage of multi-drug resistant strains in A. baumannii, Escherichia coli and P. aeruginosa. The phage acquisition rates of these six pathogens were, in descending order, Escherichia coli (80%), Stenotrophomonas maltophilia (75%), Pseudomonas aeruginosa (70%), Klebsiella pneumoniae (66.67%), Acinetobacter baumannii (36.36%) and Staphylococcus aureus (12.5%). Preliminary mNGS sequencing results showed that the pathogenic bacteria with higher abundance in the batch of effluent were Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecalis, Acinetobacter baumannii, Klebsiella aerogenes, Klebsiella michiganensis and Pseudomonas aeruginosa. In conclusion, phages of most common clinical Gram-negative pathogens were isolated from hospital wastewater with high isolation rates; however, phages of Gram-positive pathogens were isolated at lower rates, and only phages corresponding to Staphylococcus aureus were isolated in this study. The corresponding mNGS sequencing results showed that the distribution of Gram-negative pathogens in sewage may had a positive correlation with the ecological distribution of phages.

Phage therapy is one of the most important tools for the treatment of infections with multi-drug resistant bacteria. Such phages are usually isolated from hospital effluents, however, no systematic study on the distribution of phages in hospital effluents has been conducted so far. The aim of this study was to isolate the corresponding phages of common pathogenic bacteria isolated in the clinic as hosts, so as to assess the ecological distribution of phages in hospital wastewater and to provide a reference for the isolation and application of phages of drug-resistant bacteria in the clinic. A cross-sectional study design was used in this study. The 125 pathogenic bacteria (belonging to 16 different strains) isolated from the clinical microbiology laboratory of Qilu Hospital of Shandong University from May to June 2023 were selected as the target strains, and the phages corresponding to these strains were isolated and purified from the hospital wastewater by using the double-layer plate sandwich method. At the same time, the distribution of pathogenic bacteria in the same batch of wastewater was analyzed with the help of mNGS sequencing technology, so as to preliminarily investigate the abundance correspondence between pathogenic bacteria and phages in wastewater. The results showed that a total of 56 phage strains were isolated from 125 clinical pathogens as hosts, corresponding to six pathogens, including Acinetobacter baumannii, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. All six pathogenic bacteria contained strains with different degrees of drug resistance, with a higher percentage of multi-drug resistant strains in A. baumannii, Escherichia coli and P. aeruginosa. The phage acquisition rates of these six pathogens were, in descending order, Escherichia coli (80%), Stenotrophomonas maltophilia (75%), Pseudomonas aeruginosa (70%), Klebsiella pneumoniae (66.67%), Acinetobacter baumannii (36.36%) and Staphylococcus aureus (12.5%). Preliminary mNGS sequencing results showed that the pathogenic bacteria with higher abundance in the batch of effluent were Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecalis, Acinetobacter baumannii, Klebsiella aerogenes, Klebsiella michiganensis and Pseudomonas aeruginosa. In conclusion, phages of most common clinical Gram-negative pathogens were isolated from hospital wastewater with high isolation rates; however, phages of Gram-positive pathogens were isolated at lower rates, and only phages corresponding to Staphylococcus aureus were isolated in this study. The corresponding mNGS sequencing results showed that the distribution of Gram-negative pathogens in sewage may had a positive correlation with the ecological distribution of phages.

Objective To study the inhibitory effect and mechanism of citrusinol on proliferation of human hepatocellular carcinoma cells HepG2.Methods The inhibitory rate of HepG2 cells cultured in vitro was measured by MTT assay.The morphology and distribution of the ceils were observed by HE and acridine orange staining.The cell division cycle was detected by flow cytometry.The expression of F-actin protein was observed by fluorescent chromogenic method.Results Citrusinol could inhibit the growth of HepG2 cells,and the IC50 of the inhibitory rate was 76.46 μmol/L.Citrusinol could make the HepG2 cells shrink,arrest the cell division cycle to G2/M,and inhibit the expression of F-actin.Conclusion Citrusinol can prevent cell proliferation by arresting cell division cycle in G2/M phase and inhibiting the formation of cytoskeleton,thus inhibiting the growth of G2/M.

The cell origin of primary pulmonary mucinous epithelial tumors includes goblet cells, tracheobronchial mucous glands, the mucous cell metaplasia of ciliated and Clara cells, etc.There are benign, low-grade malignant potential and malignant tumors in this category. The benign tumors encompass mucous gland adenoma and mucinous cystadenoma. Ciliated muconodular papillary tumors are thought to be of low grade malignant potential or uncertain malignant potential neoplasm, while colloid adenocarcinoma and mucinous adenocarcinoma are malignant tumors. Most of primary pulmonary mucinous epithelial tumors are rare even extremely rare lesions. Similar morphological changes exist in the different tumors. Differential diagnosis for these entities may be challenging in pathological diagnosis on biopsies, even surgical sections. The clinicopathologic characteristics should be carefully analyzed to ensure accurate pathologic diagnosis for primary pulmonary mucinous epithelial tumors.

Carcinoma, Non-Small-Cell Lung ; genetics ; therapy ; China ; Consensus ; Humans ; Lung Neoplasms ; genetics ; therapy ; Molecular Targeted Therapy

Background and purpose:δ-catenin is a member of the p120 catenin subfamily, which can directly bind to E-cadherin on the cell membrane, forming E-cadherin/catenin complex. δ-catenin can also affect the cytoskeleton assembly by regulating the activity of Cdc42 (Small GTPase). Therefore, this study detected the expression of δ-catenin and Cdc42 in non-small cell lung cancer (NSCLC) and investigated the relationship between them.Methods:The expressions of δ-catenin and Cdc42 in 122 cases of NSCLC were detected by immunohistochem-istry. This study also used Western blot and real-time lfuorescent quantitative polymerase chain reaction (RTFQ-PCR) to detect the protein and mRNA expressions of δ-catenin and Cdc42 in lung cancer tissues. After up-regulating or down-regulating δ-catenin in lung cancer cell line, the activity of Cdc42 and invasion ability of lung cancer cells were detect-ed by G-LISA and Transwell.Results:The mRNA and protein expression of δ-catenin and Cdc42 in lung cancer tissues was signiifcantly higher than that in normal lung tissues. In 122 NSCLC cases, the δ-catenin positive expression rate was 65.57% (80/122), and the Cdc42 overexpression rate was 68.03% (83/122). There was a good correlation betweenδ-catenin positive expression and Cdc42 overexpression (P<0.001). The co-expression of δ-catenin and Cdc42 was related to the high clinical stage, poor differentiation, adenocarcinoma and lymph node metastasis of lung cancer (P<0.05), and was signiifcantly associated with poor prognosis in patients with lung cancer. In the lung cancer cell line, the expression and the activity of Cdc42 were changed by regulating the δ-catenin expression, which affected invasion ability of the lung cancer cells.Conclusion:The δ-catenin expression was significantly correlated with the Cdc42 expression. The co-expression of δ-catenin and Cdc42 in lung cancer was correlated with the poor prognosis of lung cancer.

Objective To investigate the mRNA and protein expression of E3 ubiquitin ligase Iduna in non-small-cell lung cancer tissue and para-neoplastic lung tissue,and the correlation of the Iduna expression with clinicopathological factors and prognosis.Methods The expression levels of the Iduna mRNA and protein in non-small-cell lung cancer tissue and para-neoplastic lung tissue were determined by reverse transcriptase-polymerase chain reaction (RT-PCR),Western-Blot and immunohistochemistry respectively,and the correlation of the Iduna expression with clinicopathological factors and prognosis was analyzed.Results RT-PCR and Western-Blot showed the expression levels of the Iduna mRNA and protein in non-small-cell lung cancer tissue (0.468 ± 0.086 and 2.554 ± 0.544) were significantly higher than those in para-neoplastic lung tissue (0.203 ± 0.070 and 1.570 ± 0.316),there were statistical differences (P ＜ 0.05).Immunohistochemistry results showed that Iduna was negative expression in the alveolar epithelium cells,negative or weak positive expression in normal bronchial and positive expression in different degrees in the non-small-cell lung cancer tissue.Iduna high expression rate was negative correlation with tumor differentiation (P =0.002),Iduna high expression rate was positive correlation with large tumor size (P =0.044),TNM staging (P=0.015) and lymph node metastasis (P=0.009).Iduna high expression of I stage non-small-cell lung cancer patients was correlated with poor post-operative survival (P =0.016).Conclusions High expression of Iduna may be related to the process of invasion and metastasis of nonsmall-cell lung cancer.It is possible that Iduna serve as potential markers for predicting prognosis in nonsmall-cell lung cancer.

Objective To observe the sedative and analgesic effects of local anesthesia with midazolam and sufent-anil for patients in ophthalmologic plastic surgery. Methods A total of 160 patients (ASA I-II) were randomly divided into two group using a random number table, 80 cases in each group, groupⅠ: local anesthesia and groupⅡ: local anesthesia with midazolam (0.05 mg/kg) and sufentanil (0.1μg/kg). Values of bispectral index (BIS), systolic blood pressure (SBP), dia-stolic blood pressure (DBP), heart rate (HR), respiratory rate (RR), oxyhemoglobin saturation(SpO2) and visual analogue scale (VAS) were recorded after patients entered into the operating room, during the time of injection of local anesthesia, at the beginning of the procedure, 20 min after the surgery and after operation. It was also recorded including hypoxemia, ap-noea, restlessness, nausea and vomiting and the duration of surgery. Results There were no significant differences in age, gender, weight and duration of surgery between two groups of patients. There was a significantly lower BIS value before inject-ing local anesthesia, at the beginning of the surgery and 20 min after the surgery compared with that of time point that pa-tients entered into the operating room in groupⅡ(P<0.05). In groupⅠthere were significantly higher values of SBP, DBP and HR during the injection of local anesthesia, at the beginning of the procedure, 20 min after the beginning of the proce-dure and after the surgery than those of time point that patients entered into the operating room;the value of RR was signifi-cantly increased during the injection of local anesthesia, at the beginning of the procedure and 20 min after the beginning of the procedure than that of time point that patients entered into the operating room (P<0.05). Compared with before anesthe-sia induction, values of SBP, DBP, HR and RR were significantly decreased in groupⅡ(P<0.05). There were significantly lower levels of SBP, DBP, HR and RR during the local anesthesia injection to the time after surgery in groupⅡthan those of groupⅠ. The value of VAS was significantly higher during the time of injection of local anesthesia and during the surgery in groupⅠthan that of groupⅡ(P<0.05). There were 15 patients with restlessness in group I, which were higher than those of groupⅡ(3 patients with restlessness). There were 3 patients with nausea and 3 patients with anoxemia and no apnea in group Ⅱ. Conclusion The conscious sedation with midazolam and sufentanil is an effective anesthetic technique for patients in ophthalmo-logic plastic surgery.

Objective To investigate the effects of different infusion rates of remifentanil infusion on spontaneous ventilation in children received sevoflurane anesthesia. Methods A total of 120 children underwent strabismus surgery were randomly assigned to four groups: C group (administration of saline), L group (remifentanil 0.03 μg · kg-1 · min-1), M group (remifentanil 0.06μg · kg-1 · min-1) and H group (remifentanil 0.09μg · kg-1 · min-1). The mean blood pressure (MBP), heart rate (HR), respiratory rate (RR), tidal volume (VT), minute ventilation (MV), endtidal CO2 [p(CO2)] and endtidal SEV were recorded after laryngeal mask insertion (T1), an initial bolus dose of remifentanil (T2),10 mins after remifentanil infu-sion (T3),15 mins after remifentanil infusion (T4) and laryngeal mask remove (T5) respectively. The adverse events and time of induction, maintenance and emergence were also recoded. Results There were no significant differences in patient age, body mass index, anesthesia time, operation time, HR and MBP at different time points between four groups. No body movement and hypoxemia were observed. The values of RR and MV at T3, T4 and T5 were significantly lower in H group than those of other three groups (P <0.05). Values of p(CO2)at T3 and T4 were significantly higher in H group than those of other three groups (P<0.05). The values of RR at T3, T4 and T5 were significantly lower in L group and M group than those of C group. The values of MV at T3 and T4 were significantly lower in L group and M group than those of C group. p(CO2)at T4 was significantly higher in L group and M group than that of C group(P<0.05), but no significant difference was found be-tween L group and M group. There was no significant difference in value of VT between four groups. Conclusion Remifent-anil infusion at a rate of 0.03~0.09μg·kg-1·min-1 could depress spontaneous ventilation in children received sevoflurane an-esthesia. The respiratory depression effect is mainly manifested by reduction of RR. It is a good option to choose 0.03~0.06μg · kg-1 · min-1 infusion to keep spontaneous ventilation and avoid severe respiratory depression according to the demand of operations in children.

PURPOSE: Fanconi anemia complementation group F (FANCF) is a key factor to maintaining the function of Fanconi anaemia/BRCA (FA/BRCA) pathway, a DNA-damage response pathway. However, the functional role of FANCF in breast cancer has not been elucidated. In the present study, we evaluated the chemosensitization effect of FANCF in breast cancer cells. METHODS: We performed specific knockdown of the endogenous FANCF in breast cancer cells by transfecting the cells with an FANCF short hairpin RNA (shRNA) vector. Cell viability was measured with a Cell Counting Kit-8, and DNA damage was assessed with the alkaline comet assay. The apoptosis, cell cycle, and drug accumulation were measured by flow cytometric analysis. Protein expression levels were determined by Western blot analysis, using specific antibodies. RESULTS: The analyses of two breast cancer cell lines (MCF-7 and MDA-MB-435S) demonstrated that the FANCF shRNA could effectively block the FA/BRCA pathway through the inhibition of Fanconi anemia complementation group D2 ubiquitination. Moreover, FANCF silencing potentiated the sensitivity of cells to mitomycin C (MMC), where combined FANCF shRNA/MMC treatment inhibited cell proliferation, induced S-phase arrest, apoptosis, and DNA fragmentation, and reduced the mitochondrial membrane potential, compared with MMC treatment alone. CONCLUSION: Taken together, this study demonstrates that the inhibition of FANCF by its shRNA leads to a synergistic enhancement of MMC cytotoxicity in breast cancer cells. These results suggest that the inhibition of the FA/BRCA pathway is a useful adjunct to cytotoxic chemotherapy for the treatment of breast cancer.
Apoptosis ; Blotting, Western ; Breast ; Breast Neoplasms ; Cell Count ; Cell Cycle ; Cell Line ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; Comet Assay ; Complement System Proteins ; DNA Damage ; DNA Fragmentation ; Fanconi Anemia ; Fanconi Anemia Complementation Group F Protein ; Membrane Potential, Mitochondrial ; Mitomycin ; RNA, Small Interfering ; Ubiquitin ; Ubiquitination