[Genomic subtyping of nosocomial transmission of carbapenem resistant Klebsiella pneumoniae].

Objective: To analyze the genomic epidemiological subtyping of carbapenem resistant Klebsiella pneumoniae (CRKP) isolated from a Third-class A hospital in Zhengzhou. Methods: From December 4, 2019 to January 10, 2020, 67 strains of CRKP were isolated from the samples submitted by the clinical departments of a Third-class A teaching hospital in Zhengzhou for microbiological testing. Multi-locus sequence typing (MLST) and carbapenem resistance genes were identified by whole genome sequencing and sequence analysis. Based on the whole genome SNP, the phylogenetic tree was constructed, and 67 CRKP strains were divided into clonal groups. The isolation ward and date of each clone group were analyzed. Results: Sixty-seven CRKP strains were classified into four MLST types (STs), of which 64 were ST11. There were 62 ST11 strains carrying blaKPC-2 gene. Based on genome-wide SNP phylogenetic tree, 64 ST11 strains were divided into four clone groups, two of which were dominant clone groups, including 33 and 27 strains respectively; the other two clone groups only contained 2 strains respectively. There was no aggregation of the dominant clones in the isolation department and date. Conclusion: Multiple clonal groups of ST11 strain carrying blaKPC-2 gene are differentiated during spreading, and they can spread in parallel and independently in the same hospital.

Molecular characteristic and pathogenicity analysis of a virulent recombinant avain infectious bronchitis virus isolated in China.

A virulent infectious bronchitis virus (IBV), designated as CK/CH/GD/QY16 (referred as QY16), was isolated from a diseased chicken farm in Guangdong province, China, in 2016. The complete genome of the strain was sequenced and analyzed. The results show that the genome of QY16 consists of 27,670 nucleotides, excluding poly (A) tail, and that its genome organization is 5' UTR-1a-1b-S-3a-3b-E-M-4b-4c-5a-5b-N-6b-3' UTR-poly (A) tail. Sequence comparison among QY16 and other IBV strains was conducted and its results demonstrate that the S1 gene of QY16 has the highest nucleotide sequence identity with that of 4/91, and the other part of its genome is highly similar to that of YX10. The results of the phylogenic analysis show that the entire genome of QY16 and most of the QY16 genes are located in the same cluster as those of YX10, except for the S1 gene which is located in the same cluster with that of 4/91. It has been further confirmed by the RDP and SimPlot analysis that QY16 is a recombinant strain deriving from YX10 (as the major parental sequence) and 4/91 (as the minor parental sequence), and that the recombination occurs in a region which includes the 3'-terminal 1b sequence (85 nt) and the 5'-terminal S1 protein gene sequence (1,466 nt). The results of the vaccination-challenge test suggest that QY16 is a nephropathogenic strain of IBV and that the vaccine strains-H120 and 4/91-cannot provide effective protection against it. These results indicate that the continuing evolution of IBV strains by genetic drift and genetic recombination may lead to IBV outbreaks even among the vaccinated chickens in China.

[Molecular epidemiology of carbapenem resistant Klebsiella pneumoniae in a hospital in Beijing].

Objective: To reveal the molecular epidemiological characteristics of Carbapenem-Resistant Klebsiella pneumoniae (CRKP) isolated from a three level teaching hospital in Beijing. Methods: Pulsed field gel electrophoresis (PFGE) was carried out to subtyping 375 CRKP isolated in that hospital between May 2010 and October 2015. Fifteen strains were chose based on the PFGE patterns to be analyzed by multi-locus sequence typing (MLST) and detection of carbapenem-resistance genes. One strain (A1502) was selected for whole genome sequencing and analyzing. Results: The 375 CRKP were divided into 140 PFGE types, among which five types contained more than five strains. The dominant types were distributed in different time periods and wards. Among the 15 strains tested by MLST and carbapenem-resistance genes detection, 13 were ST11 strains carrying KPC-2 gene. By genome-based typing, A1502 was clustered together with strains from other hospitals of Beijing but far from the strains from Shanghai and Hangzhou. Conclusion: The CRKP epidemic clone (ST11 clone carrying KPC-2) has been spreading within single hospital and across different hospitals in Beijing.

[First aid system for trauma: development and status].

With the great progress of the economy, the level of industrialization has been increasing year by year, which leads to an increase in accidental trauma accidents. Chinese annual death of trauma is already more than 400 000, which makes trauma the fifth most common cause of death, following malignant tumor, heart, brain and respiratory diseases. Trauma is the leading cause of the death of young adults. At the same time, trauma has become a serious social problem in peace time. Trauma throws great treats on human health and life. As an important part in the medical and social security system, the emergency of trauma system occupies a very important position in the emergency medical service system. In European countries as well as the United States and also many other developed countries, trauma service system had a long history, and progressed to an advanced stage. However, Chinese trauma service system started late and is still developing. It has not turned into a complete and standardized system yet. This review summarizes the histories and current situations of the development of traumatic first aid system separately in European countries, the United States and our country. Special attentions are paid to the effects of the pre- and in-hospital emergency care. We also further try to explore the Chinese trauma emergency model that adapts to the situations of China and characteristics of different regions of China. Our review also introduces the trauma service system that suits the situations of China proposed by Professor Jiang Baoguo's team in details, taking Chinese conditions into account, they conducted a thematic study and made an expert consensus on pre-hospital emergency treatment of severe trauma, providing a basic routine and guidance of severe trauma treatment for those pre-hospital emergency physicians. They also advised to establish independent trauma disciplines and trauma specialist training systems, and to build the regional trauma care system as well as the standards for graded treatment, thus establishing a multiple disciplinary team (MDT) of severe trauma. In this way, we can reduce the mortality and disability risks of severe trauma, improve the quality of patients' life, and save more lives.

Dynamics of Th17 associating cytokines in Cryptosporidium parvum-infected mice.

Cryptosporidium parvum commonly inhabits the intestinal tract of animals and humans and can cause acute watery diarrhea and weight loss. However, host immune responses to Cryptosporidium infections are not fully understood. IL-17 (also called IL-17A) is a pro-inflammatory cytokine of Th17 cells that plays a role in the host response to Cryptosporidium baileyi infection. The present study examined levels of IL-17-specific messenger RNA (mRNA) and Th17 associating cytokines in C. parvum-infected immune-suppressed BALB/c mice using real-time quantitative PCR (qPCR). Levels of IL-17 protein were determined by ELISA. The results showed that levels of IL-17 mRNA and Th17 cell-related cytokines, namely TGF-ß, IL-6, STAT-3, RORγt, IL-22, TNF-α, and IL-23, were significantly increased (P < 0.05) in gut-associated lymphoid tissue (GALT) and spleen. IL-17 protein levels in GALT were also significantly increased (P < 0.05) after infection. The present study suggested that Th17 cells play a role in host-C. parvum interaction. These results could inform future studies of the immune response against C. parvum infection in transient immunosuppressed populations.

Epithelial-mesenchymal transition biomarkers and support vector machine guided model in preoperatively predicting regional lymph node metastasis for rectal cancer.

BACKGROUND: Current imaging modalities are inadequate in preoperatively predicting regional lymph node metastasis (RLNM) status in rectal cancer (RC). Here, we designed support vector machine (SVM) model to address this issue by integrating epithelial-mesenchymal-transition (EMT)-related biomarkers along with clinicopathological variables. METHODS: Using tissue microarrays and immunohistochemistry, the EMT-related biomarkers expression was measured in 193 RC patients. Of which, 74 patients were assigned to the training set to select the robust variables for designing SVM model. The SVM model predictive value was validated in the testing set (119 patients). RESULTS: In training set, eight variables, including six EMT-related biomarkers and two clinicopathological variables, were selected to devise SVM model. In testing set, we identified 63 patients with high risk to RLNM and 56 patients with low risk. The sensitivity, specificity and overall accuracy of SVM in predicting RLNM were 68.3%, 81.1% and 72.3%, respectively. Importantly, multivariate logistic regression analysis showed that SVM model was indeed an independent predictor of RLNM status (odds ratio, 11.536; 95% confidence interval, 4.113-32.361; P<0.0001). CONCLUSION: Our SVM-based model displayed moderately strong predictive power in defining the RLNM status in RC patients, providing an important approach to select RLNM high-risk subgroup for neoadjuvant chemoradiotherapy.

Aberrant methylation of genes in stool samples as diagnostic biomarkers for colorectal cancer or adenomas: a meta-analysis.

BACKGROUND: An increasing number of hypermethylated genes in stool samples have been reported as biomarkers for the detection of colorectal cancer (CRC) or adenomas. We aimed to comprehensively review and compare the evidence for feasibility of using these biomarkers for the detection of colorectal neoplasia. METHODS: We searched Medline, the Web of Science and OVID for studies that used hypermethylated genes as biomarkers for the detection of CRC or adenomas. A meta-analysis was carried out using the random-effect model with diagnostic odd ratios (DOR) and 95% confidence intervals (CI) as effect measurements. RESULTS: A total of 19 studies including 2,356 patients were eligible for final analysis. The sensitivity, specificity, positive likelihood ratio, negative likelihood ratio and DOR for the detection of CRC or adenomas were 0.62 (95% CI: 0.51-0.71), 0.89 (95% CI: 0.86-0.92), 5.66 (95% CI: 4.68-6.83), 0.43 (95% CI: 0.34-0.55) and 13.15 (95% CI: 9.82-17.60) respectively. Of these, the sensitivity and specificity for the detection of adenoma were 0.54 (95% CI: 0.39-0.68) and 0.88 (95% CI: 0.83-0.92) respectively. CONCLUSIONS: Hypermethylated gene panels are not currently accurate enough to be used alone for colorectal neoplasia screening. The discovery and evaluation of additional biomarkers with improved sensitivity and specificity is necessary.

A multi-functional online measurement system for neuron-microelectrode interface study.

The goal of this paper is to develop a multifunctional measurement system for neuron-microelectrode interface study by LabVIEW. The system uses a commercial 60-channel microelectrode array (MEA) as an interface platform. These online functions of neuronal stimulation, extracellular potential recording, impedance monitoring, and data storage have been integrated into this system. Two offline analysis functions of wavelet de-noising and artifact removing are also included. The former two online functions are executed respectively by a NI-DAQ card and by a self-made analog frond-end amplifier, and Agilent 4284A LCR meter makes the impedance measurement. We have completed the basic functional verifications on a cultured MEA with PC-12.

Vitamin A and risk of HIV-1 seroconversion among Kenyan men with genital ulcers.

BACKGROUND: Vitamin A is involved in normal immune function and the maintenance of mucosal integrity through complex effects on cellular differentiation. OBJECTIVE: We sought to determine whether serum vitamin A levels were associated with altered susceptibility to primary infection with HIV-1 in men with high-risk sexual behaviour and genital ulcers who presented for treatment at an STD clinic in Nairobi, Kenya. METHODS: HIV-1 seronegative men were prospectively followed. Vitamin A levels at study entry were compared among 38 men who HIV-1 seroconverted versus 94 controls who remained HIV seronegative. RESULTS: Vitamin A deficiency (retinol less than 20 microg/dl) was very common and was present in 50% of HIV-1 seroconverters versus 76% of persistent seronegatives. Seroconversion was independently associated with a retinol level greater than 20 microg/dl (HR 2.43, 95% CI 1.25-4.70, P = 0.009), and a genital ulcer aetiology caused by Haemophilus ducreyi (HR 3.49, 95% CI 1.03-11.67, P = 0.04). Circumcision was independently associated with protection (HR 0.46, 95% CI 0.23-0.93, P = 0.03). CONCLUSION: Vitamin A deficiency was not associated with an increased risk of HIV-1 infection among men with concurrent STD. A decreased risk of HIV-1 seroconversion was independently associated with lower retinol levels. The effects of vitamin A on macrophage and lymphoid cell differentiation may paradoxically increase mucosal susceptibility to HIV-1 in some vulnerable individuals, such as men with genital ulcers. Lack of circumcision and chancroid are confirmed as important co-factors for heterosexual HIV-1 transmission. The role of vitamin A in heterosexual HIV-1 transmission requires further study.

Evaluation of D-xylose and 1% methyl-alpha-D-glucopyranoside fermentation tests for distinguishing Enterococcus gallinarum from Enterococcus faecium.

To determine the validity of the rapid xylose and methyl-alpha-D-glucopyranoside (MDG) fermentation tests in distinguishing Enterococcus gallinarum from Enterococcus faecium, 156 well-characterized clinical isolates of enterococci (55 E. gallinarum, 91 E. faecium, and 10 Enterococcus faecalis isolates) known to be of different clones were examined in a blinded fashion. Species identification was confirmed by PCR of the ddl ligase genes of E. faecium and E. faecalis and the vanC1 gene of E. gallinarum. Xylose tests were performed with D-xylose tablets by using a heavy bacterial suspension and were interpreted after 2 h of incubation. Standard MDG fermentation tests were read after 24 h of incubation. The xylose fermentation test had a sensitivity of 98% (54 of 55) and a specificity of 99% (100 of 101) in distinguishing E. gallinarum from E. faecium and E. faecalis. The standard MDG test had a sensitivity of 100% (55 of 55) and a specificity of 95% (96 of 101) after 24 h. The xylose fermentation test is a simple method, easily incorporated into laboratory protocols, that distinguishes E. gallinarum from E. faecium with high sensitivity and specificity in 2 h. The standard MDG test has high sensitivity and can be useful in ruling out the presence of E. gallinarum but requires overnight incubation.

Decreased susceptibility of Streptococcus pneumoniae to fluoroquinolones in Canada. Canadian Bacterial Surveillance Network.

BACKGROUND: Fluoroquinolones are now recommended for the treatment of respiratory tract infections due to Streptococcus pneumoniae, particularly when the isolates are resistant to beta-lactam antibiotics. Although pneumococci with reduced susceptibility to fluoroquinolones have been identified, their prevalence has not been determined in a defined population. METHODS: We performed susceptibility testing on 7551 isolates of S. pneumoniae obtained from surveillance in Canada in 1988 and from 1993 to 1998. Pneumococci with reduced susceptibility to fluoroquinolones (defined as a minimal inhibitory concentration of ciprofloxacin of at least 4 microg per milliliter) were further characterized. We also examined antibiotic prescriptions dispensed in Canadian retail pharmacies. RESULTS: Between 1988 and 1997, fluoroquinolone prescriptions increased from 0.8 to 5.5 per 100 persons per year. The prevalence of pneumococci with reduced susceptibility to fluoroquinolones increased from 0 percent in 1993 to 1.7 percent in 1997 and 1998 (P=0.01). Among adults, the prevalence increased from 1.5 percent in 1993 and 1994 combined to 2.9 percent in 1997 and 1998 combined. The prevalence was higher in isolates from older patients (2.6 percent among those 65 years of age or older vs. 1.0 percent among those 15 to 64 years of age, P<0.001) and among those from Ontario (1.5 percent, vs. 0.4 percent among those from the rest of Canada; P< 0.001). Fluoroquinolone use was greatest among the elderly and in Ontario. The 75 isolates (17 serotypes) of pneumococci with reduced susceptibility to fluoroquinolones were submitted by 40 laboratories in eight provinces. Reduced susceptibility to fluoroquinolones was associated with resistance to penicillin. CONCLUSIONS: The prevalence of pneumococci with reduced susceptibility to fluoroquinolones is increasing in Canada, probably as a result of selective pressure from the increased use of fluoroquinolones.

Long-term follow-up of a randomized trial of interferon therapy for chronic hepatitis B in a predominantly homosexual male population.

BACKGROUND/AIMS: Extended follow-up of a previously published therapeutic trial with interferon alfa is now available to further clarify the long-term outcome of HIV-negative and HIV-positive subjects with chronic hepatitis B virus infection after interferon alfa therapy. METHODS: Forty-five subjects with compensated liver disease and chronic hepatitis B infection with evidence of active hepatitis B replication were studied. These subjects were enrolled between 1986 and 1991 and had been randomized, stratified by HIV status, to either receive interferon therapy (10 MU/m2 of lymphoblastoid interferon alfa 3 times per week for 12 weeks) or no treatment. Hepatitis B serology, serum hepatitis B viral DNA and alanine aminotransferase were measured on an annual to biannual basis. CD4-positive T lymphocyte counts and HIV RNA concentration were also obtained. RESULTS: From 9 months post-interferon alfa treatment to the end of the extended follow-up (4 to 9 years), the relative risk of seroconverting to anti-HBe positive for subjects who had received interferon alfa therapy compared to those who did not was not significant in either HIV-negative (p = 0.80) or HIV-positive (p = 0.62) subjects. CONCLUSIONS: Unlike the first 9 months following interferon alfa therapy, the rate of elimination of markers of hepatitis B virus replication, regardless of HIV status, was not increased above the natural rate beyond 9 months following interferon alfa therapy.

Thrombotic thrombocytopenic purpura associated with ticlopidine use: a report of 3 cases and review of the literature.

Ticlopidine hydrochloride is an antiplatelet agent used for an increasing number of indications, including cerebrovascular disease, unstable angina, coronary artery stenting, and peripheral vascular bypass grafting. It has uncommon but severe hematologic effects, including thrombotic thrombocytopenic purpura. We report 3 new cases of ticlopidine-associated thrombotic thrombocytopenic purpura and review the English-language literature. Of the 13 patients described (10 from published articles), an equal number were women and men. The median age of the women was 50 years, and that of the men was 72 years. Thrombotic thrombocytopenic purpura occurred within 2 to 8 weeks of starting ticlopidine therapy. Survivors received plasma therapy, but of the 4 who died, 3 had received platelet transfusions. With discontinuation of the drug and prompt plasma exchange therapy, mortality was comparable to that seen with idiopathic thrombotic thrombocytopenic purpura, and relapse was uncommon. Physicians and patients should be aware of this potentially fatal but treatable complication of ticlopidine therapy.

Sulbactam/cefoperazone versus cefotaxime for the treatment of moderate-to-severe bacterial infections: results of a randomized, controlled clinical trial.

We conducted a randomized, open-label, controlled, multicenter study to compare sulbactam/cefoperazone with cefotaxime in terms of efficacy and safety for the treatment of hospitalized patients with moderate-to-severe bacterial infections. More than two-thirds of the pathogens recovered from these patients produced beta-lactamase. Two hundred-seven (88.1%) of the 235 patients enrolled completed the study and were included in the efficacy and safety evaluations. One hundred-three patients received sulbactam/cefoperazone (2-4 g/d) administered in evenly divided doses every 12 hours by a 30-minute intravenous drip; 104 patients received cefotaxime (6-12 g/d) administered in evenly divided doses every 6 or 8 hours by a 30-minute intravenous drip. The overall efficacy rates (i.e., cure or markedly improved) were 95% for the sulbactam/cefoperazone group and 90% for the cefotaxime group (P = .186), whereas the bacterial eradication rates were 85% for the sulbactam/cefoperazone group and 81% for the cefotaxime group (P = .467). Both drug regimens were well tolerated. Sulbactam/cefoperazone is effective and safe for the treatment of moderate-to-severe bacterial infections caused mainly by beta-lactamase-producing organisms.

Improved long term survival after intracavitary interleukin-2 and lymphokine-activated killer cells for adults with recurrent malignant glioma.

BACKGROUND: The median survival for adults with glioblastoma multiforme (GBM) is 12 months, despite surgery, radiation, and chemotherapy. Regimens using interleukin-2 (IL-2) plus lymphokine-activated killer (LAK) cells have been beneficial against systemic cancers, albeit with significant toxicity. METHODS: Nineteen adults with recurrent malignant glioma (5 GBMs, and 4 anaplastic astrocytomas (AA)), Karnofsky performance status 60 or greater, were treated with intracavitary autologous LAK cells plus IL-2 after reoperation. Lymphokine-activated killer cells and IL-2 were given on day 1, and IL-2 alone was given 5 times during a 2-week cycle. This cycle was repeated at 2 weeks to constitute one 6-week course of therapy. Each two-cycle course of treatment was repeated at 3-month intervals for patients with stable disease or response to therapy. At the conclusion of immunotherapy, all patients were offered chemotherapy, generally carmustine or procarbazine, including responders. Corticosteroids were strictly limited during immunotherapy. Sequential reservoir aspirates were obtained for microbiologic and cytologic analyses. RESULTS: The maximal tolerated dose for a 12-dose course of therapy was 1.2 million international units (MIU) per dose. Dose-limiting, cumulative IL-2-related central nervous system (CNS) toxicity was observed at 2.4 MIU per dose. Three responses were confirmed by computed tomography scan during therapy: one complete response (CR) (1 AA), and two partial responses (PR) (2 GBM); as well as a significant increase in GBM survival. One additional CR (GBM) was observed at 17 months. The median survival for immunotherapy patients with GBM was 53 weeks after reoperation (N = 15) (mean, 87.9 +/- 21.4 weeks, standard error for the mean), with 8 of 15 surviving more than 1 year (53%). The median survival for 18 contemporary patients with GBM reoperated and treated with chemotherapy was 25.5 weeks (mean, 27.4 +/- 3.7 weeks), with 1/18 alive at 1 year (> 6%). Six of the 15 patients with GBM had additional surgery or biopsy, and chemotherapy after immunotherapy. The contribution of subsequent chemotherapy to survival cannot be discounted. CONCLUSIONS: Lymphokine-activated killer cells and IL-2 can be administered safely within the CNS resulting in improved long term survival in patients with recurrent glioblastoma. Increased survival was associated with significant biologic changes characterized by a regional eosinophilia, and extensive lymphocytic infiltration. A prospective randomized clinical trial is warranted.