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.

Drug-associated reward memories are conducive to intense craving and often trigger relapse. Simvastatin has been shown to regulate lipids that are involved in memory formation but its influence on other cognitive processes is elusive. Here, we used a mass spectrometry-based lipidomic method to evaluate the impact of simvastatin on the mouse brain in a cocaine-induced reinstatement paradigm. We found that simvastatin blocked the reinstatement of cocaine-induced conditioned place preference (CPP) without affecting CPP acquisition. Specifically, only simvastatin administered during extinction prevented cocaine-primed reinstatement. Global lipidome analysis showed that the nucleus accumbens was the region with the greatest degree of change caused by simvastatin. The metabolism of fatty-acids, phospholipids, and triacylglycerol was profoundly affected. Simvastatin reversed most of the effects on phospholipids induced by cocaine. The correlation matrix showed that cocaine and simvastatin significantly reshaped the lipid metabolic pathways in specific brain regions. Furthermore, simvastatin almost reversed all changes in the fatty acyl profile and unsaturation caused by cocaine. In summary, pre-extinction treatment with simvastatin facilitates cocaine extinction and prevents cocaine relapse with brain lipidome remodeling.
Animals ; Brain ; Cocaine ; Conditioning, Operant ; Extinction, Psychological ; Lipidomics ; Male ; Mice ; Simvastatin/therapeutic use*

Drug-associated reward memories are conducive to intense craving and often trigger relapse. Simvastatin has been shown to regulate lipids that are involved in memory formation but its influence on other cognitive processes is elusive. Here, we used a mass spectrometry-based lipidomic method to evaluate the impact of simvastatin on the mouse brain in a cocaine-induced reinstatement paradigm. We found that simvastatin blocked the reinstatement of cocaine-induced conditioned place preference (CPP) without affecting CPP acquisition. Specifically, only simvastatin administered during extinction prevented cocaine-primed reinstatement. Global lipidome analysis showed that the nucleus accumbens was the region with the greatest degree of change caused by simvastatin. The metabolism of fatty-acids, phospholipids, and triacylglycerol was profoundly affected. Simvastatin reversed most of the effects on phospholipids induced by cocaine. The correlation matrix showed that cocaine and simvastatin significantly reshaped the lipid metabolic pathways in specific brain regions. Furthermore, simvastatin almost reversed all changes in the fatty acyl profile and unsaturation caused by cocaine. In summary, pre-extinction treatment with simvastatin facilitates cocaine extinction and prevents cocaine relapse with brain lipidome remodeling.

Objective To explore the clinical value of CEUS guided biopsy of lymphoma in anterior mediastinum.Methods The data of 36 patients with lymphoma of anterior mediastinum underwent biopsy guided by CEUS and 36 patients by conventional ultrasound retrospectively.The successful rate of biopsy and rate of complication occurence were compared between the CEUS group and conventional ultrasound group.Results The successful rate of biopsy in CEUS group was 100％ (36/36),including 26 non-Hodgkin's lymphoma (NHL),10 Hodgkin's lymphoma (HL).The total times of puncture were 60 in 36 patients.The rate of complication occurrence was 11.11 ％ (4/36).The successful rate in conventional ultrasound group was 88.89％ (32/36),including 22 NHL,14 HL.The times of puncture were 91 in 36 patients.The rate of complication occurrence was 41.67％ (15/36).There were significant differences in successful rate and the rate of complication occurrence between two groups (x2 =4.235,8.651,P=0.040,0.003).Conclusion CEUS can reflect the microcirculation of lymphomas in anterior mediastinum,and can guide targeted biopsy.It can improve the successful rate of biopsy and reduce the complications.

Objective To investigate the mechanism related to reduced antibiotic sensitivity of Acinetobacter baumannii inducted by meropenem in vitro.Methods Three strains of clinically isolated carbapenems-sensitive Acinetobacter baumannii were induced by meropenem in vitro, and the mutant strains (MS1, MS2 and MS3) were obtained.Minimal inhibitory concentrations (MICs) of antimicrobial agents to strains before and after induction were determined by automatic drug sensitivity analyzer .The homology of strains was analyzed by Enterobacterial repetitive intergenic consensus -polymerase chain reaction ( ERIC-PCR).Modified Hodge test and EDTA-Na2-double disk synergy test were used to detect carbapenemase and metallo-β-lactamase (MBL), respectively.Main carbapenemase genes were detected by PCR and followed by DNA sequencing.Expressions of adeB and outer membrane proteins in strains before and after induction were detected with fluorescence quantitative PCR and SDS -polyacrylamide gel electrophoresis , respectively.t test was used for data analysis .Results The sensitivity of mutant Acinetobacter baumannii strains to meropenem and most antibiotics was reduced , except to imipenem, amikacin and polymyxin; and the reduced sensitivity to meropenem in MS2 and MS3 was of genetic stability.ERIC-PCR showed 100%homology between the mutant strains and parental strains .Both carbapenemase and metallo -β-lactamase were negative in mutant strains and parental strains , and only OXA-51 gene was found.The expressions of adeB gene in mutant strains were 24.26 ±0.91, while those in parental strains were 22.81 ±0.38, and the difference was not significant (t =2.534, P >0.05).Outer membrane protein with molecular weight 54 000 was missing in MS1, while that with molecular weight 47 000 was missing in MS2 and MS3.Conclusion Reduced antibiotics sensitivity in meropenem -induced Acinetobacter baumannii may be correlated with the deficiency of outer membrane protein with molecular weight 47 000.

OBJECTIVE: To investigate osteopontin (OPN) expression in plasma and tissue of patients with layngeal squamous cell carcinoma and analyze its role in invasion, metastasis, and clinical significance in laryngeal quamous cell carcinoma. METHOD: Enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry were used to detect expression of OPN in plasma and tissue of 60 cases of laryngeal squamous cell carcinoma, 20 cases of adjacent normal laryngeal tissue and 20 cases of plasma from healthy subjects. RESULT: The expression of plasma OPN was closely correlated with clinical stage and cervical lymphatic metastasis in laryngeal squamous cell carcinoma (P < 0.05), but no significant correlation with the tumor location, pathological grade, gender and age (P > 0.05). The expression of OPN increased in plasma during cancer development: laryngeal squamous cell carcinoma (38.089 ± 9.225) ng/ml, healthy subjects (18.563 ± 9.308) ng/ml. There was a significant difference between the groups (P < 0.05). The expression of OPN in tissue was closely correlated with clinical stage (P < 0.05), pathological grade (P < 0.05) and cervical lymphatic metastasis (P < 0.05) in laryngeal squamous cell carcinoma adjacent atypical hyperplastic epithelium and carcinoma. The expression of OPN increased in tissue during cancer development: laryngeal squamous cell carcinoma (56.67%), adjacent normal laryngeal tissue (15.00%). There was a significant difference between the groups (P < 0.05). Elevated expression of plasma OPN is positively correlated with the expression of OPN in tissue in laryngeal squamous cell carcinoma patients (r = 0. 871, P < 0.05). CONCLUSION OPN plays an important role in the infiltration, metastasis and carcinogenesis in laryngeal squamous cell carcinoma. Combination of serum OPN, tissue OPN detection can be used as diagnostic and surveillance indicators for laryngeal squamous cell carcinoma infiltration and metastasis.
Carcinoma, Squamous Cell ; metabolism ; pathology ; Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Head and Neck Neoplasms ; metabolism ; pathology ; Humans ; Hyperplasia ; pathology ; Immunohistochemistry ; Laryngeal Neoplasms ; metabolism ; pathology ; Larynx ; pathology ; Lymphatic Metastasis ; Neck ; Osteopontin ; metabolism ; Squamous Cell Carcinoma of Head and Neck

Objective To investigate the effects of 20％ fluor-hydroxyapatite(FHA) on proliferation and osteogenic differentiation of human MG63 osteosarcoma cells.Methods FHA was prepared by chemical precipitation method,and its structure and surface features were tested by scanning microscope,Xray diffraction(XRD) and Fourier transform infrared spectroscopy.MG63 cells were cultured and divided into FHA,hydroxyapatite(HA) and control groups(n=3).The proliferation of the cells was evaluated using methylthiazol tetrazolium(MTT) assay.ALP activity of the cells was assessed.Osteogenic differentiation was evaluated based on the reverse transcription PCR(RT-PCR) of differentiation-related genes,namely,collagen type Ⅰ(Col Ⅰ),alkaline phosphatase(ALP),osteocalcin(OCN) and core binding factor α1(Cbfα1).The data were analyzed statistically by one-way analysis of variance using SPSS 13.0 software.Results XRD test showed that the main crystalline phase of FHA was similar to that of HA.Absorptance value of cells exposed to FHA(1.87±0.06) measured by MTT was higher than that of the control(1.25±0.02) on the third day(P＜ 0.05),and there was no statistically significant difference between the cells exposed to FHA and HA(1.84± 0.03)(P＞0.05).ALP activity of the cells exposed to FHA (4.62±0.09) was higher than that of the control (1.92 ± 0.05)(P＜0.05).RT-PCR tests showed that compared with the control,FHA up-regulated the expression of Col Ⅰ,ALP and OCN mRNA,down-regulated the expression of Cbfα1 mRNA.Conclusions FHA enhances the proliferation and osteogenic differentiation-related gene expression,and has good biocompatibility.

Objective To investigate the causes and treatments of severe complications after laparoscopic surgery. Methods Clinical data of 7 cases with severe complications after laparoscopic surgery in the Third People’s Hospital of Shenzhen from January 2008 to January 2013 were analyzed retrospectively. The informed consents of all patients were obtained and local ethical committee approval had been received. There were 3 males and 4 females with the age ranging from 6 to 67 years old and median of 40 years old. The diagnosis, treatments clinical data of the patients were collected, and the causes, treatments and prevention measures of the complications were summarized. Results Five cases underwent laparoscopic cholesystectomy (LC), 4 of which suffered bile duct injury for severe abdominal adhesion and unclear anatomy of the Calot triangle and 1 case suffered strangulated intestinal obstruction after operation because the hepatic falciform ligament was ruptured when puncturing below the xiphoid. One case underwent laparoscopic high ligation of hernial sac for indirect inguinal hernia and suffered strangulated intestinal obstruction after operation because part of the small intestine ran out of the peritoneum through the port site below the umbilicus. One case underwent laparoscopic tension-free repair for indirect inguinal hernia and suffered ischemic necrosis of the left testis possibly because of testicular torsion when stretching the tissue. The 4 cases with bile duct injury received primary drainage, 3 of which underwent secondary Roux-en-Y cholangiojejunostomy for severe biliary stricture and all recovered well after operation, and the other case underwent endoscopic biliary stent implantation 3 months after operation. The 2 cases with strangulated intestinal obstruction underwent emergent partial resection and end-to-end anastomosis of small intestine and recovered well after operation. The case with necrosis of testis underwent testicular excision. Conclusions The main causes of severe complications after laparoscopic surgery probably are the unclear local anatomy and failing to convert surgical procedure timely, etc. Early diagnosis, comprehensive evaluation and proper surgical interventions are the important measures for improving the prognosis of these patients.

Objective To investigate molecular typing and drug resistance patterns of 98 Klebsiella pneumoniae (K.pneumoniae) isolated from type 2 diabetes patients complicated with maxillofacial infection,to research the virulence and resistance mechanisms.Methods The study was a prospective study that adopted the method of continuous sampling from fixed location,from March 2010 to October 2012.The maxillofacial surgery patients diagnosed with type 2 diabetes complicated with maxillofacial infection were chosen in 7 hospitals in Zhengzhou as the research object,and a total of 431 pus sample were collected continuously,in which 98 strains K.pneumoniae were isolated and identified.The Kirby-Bauer disk diffusion test was conducted in 98 strains to determine the resistance to 19 antibacterial agents.K.pneumoniae chromosomal DNA were digested by restriction endonuclease Xba Ⅰ and analyzed by pulsed-field gel electrophoresis (PFGE).PFGE patterns of K.pneumoniae strains were analyzed using Fingerprinting software.The relationship between the molecular types and resistance phenotype was observed.The extended spectrum β-1actamase-producing K.pneumoniae were screened out by the double disc synergy test (DDST)Polymerase chain reaction (PCR) was used to detect resistant genotypes,serotype and virulence genes.The purified PCR products of resistant genes were cloned and sequenced.Hypermucoviscosity phenotype of all strains were determined by string test.Results Much severer drug-resistance for K.pneumoniae was identified and the result of extended-spectrum beta-lactamase producing rate was 57.1 ％.Ninety-eight strains were dispatched into 13 groups by PFGE.No dominant bands and specific extended-spectrum beta-lactamase DNA bands were found.The results of PCR showed that among the 56 strains of extended spectrum β-lactamase-producing K.pneumoniae,40 were positive for blaSHV (accounting for 71.4％),28 positive for blaTEM (accounting for 50.5％),21 positive for blaCTX-M (accounting tor 37.5％).The sequencing results were as follows:TEM-1,CTX-M-3 and a variety of SHV.Serotype K1,K2,K3,K5,K20,K54 and K57 and 3 kinds of virulence genes were detected,but not in strong toxicity-based.Hypermucoviscosity positive rate was 31.6％ (31/98).Conclusion Much severer drug resistance of K.pneumoniae in this study was identified and resistant mechanism was complex,in which strong toxicity serotype and virulence geues exist,which need more attention from clinical.