Objective To perform single-molecule, real-time sequencing of ceftazidime-avibactam (CAZ-AVI)-resistant Pseudomonas aeruginosa and to investigate the mechanism underlying ceftazidime-avibactam resistance in P. aeruginosa. Methods The susceptibility of 89 P. aeruginosa isolates randomly sampled from clinical specimens in Sanming First Hospital Affiliated to Fujian Medical University from November 2021 through July 2023 to common antimicrobial agents was tested, and the minimum inhibitory concentration (MIC) of CAZ-AVI was determined against P. aeruginosa with a broth microdilution assay, with CAZ-AVI MICs of 8 mg/L and lower defined as susceptible and 16 mg/L and higher as resistant. The expression of drug-resistant genes ampC, oxa-488, oprD, mexA, oxa-10, oxa-14, vim and tem was quantified in P. aeruginosa using a real-time quantitative reverse transcription PCR (qPCR) assay. CAZ-AVI-susceptible and -resistant P. aeruginosa isolates from the same case were selected for PacBio single-molecule, real-time sequencing, and sequencing results were subjected to genome structure and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional annotations. Results The 89 P. aeruginosa isolates showed a relatively high level of resistance to meropenem (75.28%) and imipenem (74.16%) and the highest susceptibility to amikacin (91.01%). There were 49 CAZ-AVI-resistant P. aeruginosa isolates and 40 susceptible isolates. qPCR assay detected lower oprD gene expression in CAZ-AVI-resistant P. aeruginosa isolates [0.104 (2.385)] than in susceptible isolates [0.551 (17.885)] (Z = -2.958, P < 0.01), and there were no significant differences between CAZ-AVI-susceptible and -resistant P. aeruginosa isolates in terms of ampC, oxa-488, mexA or tem gene expression (all P values > 0.05), while oxa-10, oxa-14 and vim gene was expressed in few P. aeruginosa isolates. There were 1 729, 3 936, 3 737 and 3 955 genes in CAZ-AVI-resistant P. aeruginosa isolates PA-762 and PA-M174 and susceptible isolates PA-885 and PA-808 that were annotated to GO terms, with the highest numbers of genes enriched in the molecular function of catalytic activity, high numbers of genes enriched in biological processes of metabolic process, single-organism process and cellular process, and high numbers of genes enriched in cellular components of cell and cell membranes. There were 1 803, 4 084, 3 915 and 4 066 genes in the PA-762, PA-M174, PA-885 and PA-808 isolates enriched in the KEGG signaling pathway, and the majority of genes were enriched in four primary signaling pathways of metabolism, genetic information processing, environmental information processing and cellular process, with the highest number of genes associated with metabolic pathways. Both CAZ-AVI-resistant P. aeruginosa isolates PA-762 and PA-M174 carried multiple efflux pumps systems, including MexAB-OprM, MexCD-OprJ, MexEF-OprN and MexXY-OprM. Single nucleotide substitution was found at position 169 in the DNA sequence of the PA-762 isolate, leading to substitution of serine for glycine at position 57 in the protein sequence, and there are deletions of two bases at positions 307 and 308 in the DNA sequence of the PA-M174 isolate, leading to substitution of threonine for arginine at position 103 in the protein sequence. Conclusion Mutation or downregulation of oprD gene may lead to CAZ-AVI resistance in P. aeruginosa.

In order to deal with the shortage of medical talents in Tibet, medical group assistance to Tibet is started as an innovative move in a new era, not only providing high-quality medical resources to Tibet, but also training local medical personnel by drawing on the "Master-Apprentice" model in traditional Chinese medicine. During the past three-year, the Shanghai Medical Team took advantage of medical group assistance to Tibet, enriched the types, methods and contents of teaching and mentoring tasks, and highlighted the role of "experts leading the backbone" and "team leading the team" in the "Master-Apprentice" model. The total amount and quality of local medical talents have thus been significantly improved. On the basis of summarizing experience, this study proposes a number of measures to optimize the current "Master-Apprentice" training model, evaluate the implementation process, and improve the feedback and quality management, so as to speed up the construction of the Tibetan medical talent team.

Objective:To investigate effects of the autophagy inducer rapamycin and autophagy inhibitor 3-methyladenine on viral structures and biosynthesis of functional proteins in dorsal root ganglia in a guinea pig model of varicella-zoster virus (VZV) infection, and to explore their possible mechanisms.Methods:VZV was cultured and proliferated in human embryonic lung fibroblasts (HELFs) , and peripheral blood mononuclear cells (PBMCs) were isolated from guinea pigs. VZV-HELFs and PBMCs were co-cultured for 18-20 hours, and VZV-PBMCs were collected by centrifugation. Thirty-two guinea pigs were randomly and equally divided into 4 groups (8 mice in each group) : blank control group was injected with autologous PBMCs via the medial canthal venous plexus; autophagy inhibition group, autophagy induction group, and VZV infection group were intraperitoneally injected with 3 mg/kg 3-methyladenine solution, 0.5 mg/kg rapamycin solution, and the same volume of 0.9% NaCl solution respectively, followed 2 hours later by injections with 50 μl of VZV-PBMCs via the medial canthal venous plexus. Fourteen days later, the guinea pigs in each group were sacrificed, and dorsal root ganglion tissues were collected. The transmission electron microscope was used to observe the morphology of virus particles, as well as the morphology and number of autophagic vesicles, Western blot analysis was performed to determine the expression of VZV nucleocapsid protein (NCP) , immediate-early protein 62 (IE62) , and autophagy-related proteins Beclin-1 and p62, and immunohistochemical study to determine the expression of anti-VZV antibodies in VZV-infected dorsal root ganglia. Statistical analysis was carried out by using two-independent-sample t test, one-way analysis of variance, least significant difference- t test or Kruskal-Wallis H test. Results:Nucleocapsid-containing virions and scattered autophagosomes were seen in the dorsal root ganglia in the VZV infection group under the transmission electron microscope. The number of autophagic vesicles significantly differed among the blank control group, VZV infection group, autophagy induction group and autophagy inhibition group ( M[ Q1, Q3]: 0, 5[4, 6], 7[5, 9], 0, respectively; H = 135.60, P < 0.01) , and was significantly higher in the VZV infection group than in the blank control group and autophagy inhibition group (both P < 0.05) , as well as in the autophagy induction group than in the autophagy inhibition group ( P<0.05) , but there was no significant difference between the VZV infection group and autophagy induction group ( P>0.05) . Western blot analysis showed that the expression level of IE62 protein was significantly higher in the VZV infection group (1.49 ± 0.06) than in the blank control group (0.50 ± 0.09, t = 9.17, P < 0.05) ; the expression of anti-VZV antibodies was significantly lower in the autophagy inhibition group than in the autophagy induction group and VZV infection group ( t = 9.24, 7.78, respectively, both P < 0.01) , while there was no significant difference between the autophagy induction group and VZV infection group ( P > 0.05) . Conclusion:Autophagy occurred in the dorsal root ganglia of guinea pigs after VZV infection; the inhibition of autophagy could affect the structure of VZV and decrease the expression of VZV functional proteins in the dorsal root ganglia of guinea pigs.

Objective To evaluate the curative effect of recombinant human Interferon α2b (rhIFN α2b) spray in hand,foot and mouth disease (HMFD).Methods In total,313 HMFD children were enrolled from Beijing Ditan Hospital affiliated to Capital Medical University,Tianjing Second People's Hospital,Hebei Children's Hospital,the Second Hospital affiliated to Wenzhou Medical College,Kunming Maternal and Child Health Hospital and Guiyang Public Health Treatment Center from March,2015 to February,2017.They were divided into rhIFN α2b group (148 cases) and ribavirin group (165 cases).The children in rhIFN α2b group were given with the rhIFN α2b spray,and those in ribavirin group were given with the ribavirin spray.Meanwhile,the children were given unified standard interventions for basic treatment.The curative effect and safety between two groups was compared.The t test was used for intergroup comparison and the Wilcoxon rank test was used for non-normal quantitative data.Results At the end of the follow-up period,all kids reached the recovery level,with 144 cases in the rhIFN α2b group and 164 cases in the ribavirin group.Fever,herpes and rashes all disappeared with 7-day follow-up.The total efficiency of the rhIFN α2b group measured at the 72h after treatment was 74.15％,which showed significant differences compared with the ribavirin group with 49.09 ％ efficiency (Z=4.44,P＜0.01).As the secondary outcome measures,the complete disappearance time of fever and the immediate disappearance time of fever in the rhIFN α2b group were significantly shorter than those in the ribavirin group ([27.03±21.99] vs [33.21±26.71],t=-2.13;[23.56±13.96] vs [28.51±18.84],t=-2.07,both P＜0.05).The appetite improvement and the disappearance times of oral herpes and rashes in the rhIFN α2b group were shorter than those in the ribavirin group,with significant differences (x2 =4.94,3.17 and 3.55,respectively,all P=0.000).No adverse event in both groups.Conclusions rhIFN α2b spray treatment in HMFD is proved significantly effective,particularly,it can evidently relieve fever symptoms and promote the disappearance of oral herpes symptoms,and no adverse event is observed throughout the study,which indicates a good safety of the rhIFN α2b spray.Clinical trial registration Chinese Clinical Trial Registry,ChiCTR-OIN-17013182.

ObjectiveTo evaluate the feasibility of determining the chemical composition of kidney stones using gemstone spectral imaging ( GSI ).Methods One hundred and sixty eight extracted human kidney stones immersed in a 10 cm deep water tank underwent CT (Discovery CT750 HD) scans with GSI mode and conventional polychromatic imaging ( CPI,120 kVp) mode.All GSI data were transferred to Workstation AW 4.4 to acquire monochromatic images of 50 keY,effective atomic number (Zeff) mapping images,water (calcium)-based images and calcium (Water)-based images with GSI Viewer.CT numbers of stones were measured and compared at 50 keV monochromatic images and 120 kVp polychromatic images,the mean Zeff,calcium density and water density were measured at Zeff mapping images,Calcium (Water) -based images and Water (Calcium)-based images,respectively.The mean Zeff,spectral HU curve slope and calcium water ratio (CWR) were compared with ANOVA and Wilcoxon test.The composition of kidney stones was determined by infrared spectrometer after CT examination.According to the result of stone composition determined by infrared spectroscopy,108 pure kidney stones were divided into five groups:Uric acid stones ( UA,n = 13 ),struvite stones ( STR,n = 24),cystine stones ( CYS,n = 14),calcium phosphate stones ( CaP,n = 18),and calcium oxalate stones ( COX,n = 39).ResultsThe mean Zeff,CWR,the mean CT numbers at 50 keV images,120 kVp images and spectral HU curve slope of each group were listed as the following:UA [ 7.4 ± 0.4,0.0085 ± 0.0021,( 503 ± 168 ) HU,(495 ± 106 ) HU and - 0.77 ] ; STR [ 11.8 ± 0.9,0.1743 ± 0.0677,( 1056 ± 290 ) HU,( 799 ± 165 ) HU and 18.72 ] ; CYS [ 11.2 ± 0.6,0.1253 ± 0.0297,( 740 ± 172 ) HU,( 565 ± 129 ) HU and 12.79 ] ; CaP [ 16.0 ± 0.4,0.6781 ± 0.0952,( 2567 ±178 ) HU,( 1602 ± 200 ) HU and 37.14 ] ; COX [ 15.4 ± 0.4,0.5683 ± 0.0759,( 2267 ± 385 ) HU,( 1489 ±284) HU and 36.36 ],there were significant differences among groups ( P ＜ 0.01 ).The differences in the mean Zeff,CRW,spectral HU curve slope were statistically significant among the five groups ( P ＜ 0.05 ).Conclusion Spectral CT imaging provides a new method to characterize the kidney stones with the information orovided by mean Zeff,CRW and the CT numbers at 50 keV.

OBJECTIVETo investigate the involvement of immunoreactive-dynorphin A in the inhibitory effect of N-nitro-L-arginine on the morphine physical dependence in rats.

METHODSThe rats were rendered dependent on morphine by subcutaneous administration of morphine solution three times daily in a manner of dose increment of 5 mg.kg(-1) for 6 days. The degree of morphine physical dependence was monitored by scoring the abstinence syndromes precipitated by 5 mg.kg(-1) naloxone of the rats. The expression levels of immunoreactive dynorphin A in tissues were determined using a radioimmunoassay.

RESULTSIntraperitoneal injection of 5 mg.kg(-1) N-nitro-L-arginine suppresses most of the withdrawal symptoms of morphine dependent rats. N-nitro-L-arginine can elevate the expression of immunoreactive dynorphin.

CONCLUSIONSChronic N-nitro-L-arginine administration can inhibit the development of morphine physical dependence in a manner of dose-dependence, which is significantly related to its role of regulating the endogeneous dynorphin system.

Animals ; Dose-Response Relationship, Drug ; Dynorphins ; physiology ; Male ; Morphine Dependence ; prevention & control ; Nitroarginine ; pharmacokinetics ; pharmacology ; Rats ; Rats, Sprague-Dawley

Objective To investigate the role of tumor suppressor gene PTEN in colorectal cancer. Methods Using Northern blot,immunohistochemistry, colorectal carcinoma was examined in 47 cases. ResultsKG1 The expression of PTEN mRNA in colorectal cancer was lower than that in paired para-carcinoma tissues( P

Objective:To study the effect of gypenosides (GPs) on the hepatic function and fibrosis progression in Wistar rats with immune fibrosis. Methods: The model of immune fibrosis was established by injection of human albumin. Rats in treated group were also administered with 0. 5 mg GPs during 30 d human albumin attack. The hepatic function of rats was e-valuated by determining the levels of ALT, TBIL and TBA in serum. The progression of hepatic fibrosis was evaluated by the levels of HA, PC Ⅲ and LN by radioimmunoassay (RIA) and confirmed by rats tissue pathology analysis. Results: Albumin injection significantly elevated the levels ALT, TBIL, TBA, HA, PCⅢ(P

Objective To investigate the effect of N-nitro-L-arginine (NO2Arg) on the development of morphine tolerance and dependence.Methods Forty male SD rats weighing (180 ? 18)g were randomly divided into 5 groups for the study of morphine physical dependence and tolerance : group Ⅰ normal saline; group Ⅱ morphine; group Ⅲ morphine + 1 mg?kg-1 NO2Arg; group Ⅳ morphine + 5 mg?kg-1 NO2 Arg ; group Ⅴ morphine +20 mg?kg-1 NO2Arg. Twenty-four male Kunming mice weighing (20.0 ? 2.3)g were randomly divided into 3 groups for the study of psychological dependence: group Ⅵ morphine; group Ⅶ dihydro etorphine; group Ⅷ normal saline. The morphine physical dependence was induced by dose increment. The degree of morphine tolerance was assessed by the pain threshold to heat radiation using tail-flick test. The degree of morphine physical dependence was determined by evaluating the abstinence syndrome precipitated by 4 mg?kg -1 naloxone. The psychological dependence of mice was evaluated by conditioned place preference. Results Consecutive injection of NO2Arg inhibited the development of morphine tolerance in a dose-dependent manner. 5 mg?kg-1 NO2 Arg suppressed most abstinent symptoms of morphine dependent rats. The duration when morphine and dihydro etorphine psychologically dependent mice stayed in the preferred compartment increased significantly from (6.1? 2.0)min and (8.0?0.7)min to (9.3?1. l)min and (9.5 ? 1.2) min by 4 mg?kg-1 NO2 Arg. Conclusion NO2 Arg can inhibit the development of morphine tolerance and dependence and reverse the existing psychological dependence.