Objective To investigate the status of genotype of the KPC(Klebsiella pneumoniae carbapenemase)-encoding genes in Pan-resistant K. Pneumoniae, isolated from the 98th Hospital of People' s Liberation Army, Huzhou district, Zhejiang province, China. Methods 19 strains of Pan-resistant K. Pneumoniae were isolated from the inpatients between November, 2008 and July,2009. Phenotypic confirmatory test for suspected carbapenemases production were carried out by Modified Hodge test. Carbapenemase gene of blaKPC was analyzed by PCR and verified by DNA sequencing. Results In 19 strains of K. Pneumoniae, the positive rates of Modified Hodge test and gene of blaKPC were both 100.0%. These genes all belonged to blaKPC-2 subtype confirmed by nucleotide sequence analysis. Among them, the blaKPC-2 gene sequence of the HZ001 strain (its original serial number was HZ9871 ) had been registered in GenBank (GenBank Accession Number: GU086225).Conclusion All of the Pan-resistant K. Pneumoniae isolated from the inpatients harbored blaKPC-2 type carbapenemases gene and causing an outbreak in a hospital. Carbapenemases that producing type KPC-2 might be the major reason which causing the resistance to Carbapenems antibiotics.

Objective:To investigate the epidemiology, evolution and transmission characteristics of HIV-1 CRF07_BC in Nanjing between 2015 and 2019 to provide scientific basis for precise prevention and control of HIV-1 transmission.Methods:Pol gene sequences of 319 patients with HIV-1 CRF07_BC infection in Nanjing were amplified and sequenced and a maximum likelihood phylogenetic tree was then constructed. Markov Chain Monte Carlo sampling method was used to generate Maximum Clade Credibility Tree. Bayesian Skyline Plot was used to reconstruct the changing trend of the effective population size. Pairwise gene distance method was used to construct molecular network to investigate the transmission features. Results:Among the 319 patients, 303 (95.0%) were male; 264 (82.8%) had multiple sexual partners; only 14 (4.4%) had been using condoms. Most of the infections were occurred in men who have sex with men (MSM), accounting for 77.4%. The maximum likelihood phylogenetic analysis of HIV-1 CRF07_BC revealed two clusters: Cluster1 and Cluster2. Cluster1 mainly contained the strains isolated from MSM and Cluster2 mainly consisted of the strains isolated from heterosexual patients. The most recent common ancestor was 2002.47(1999.91, 2005.43) year for Cluster1 and 1996.38(1992.55, 1999.76) year for Cluster2. The evolutionary rates (95% highest posterior density, 95%HPD) of Cluster1 and Cluster2 were 1.73×10 -3 (1.36×10 -3-2.16×10 -3) substitutions·site -1·year -1 and 2.09×10 -3 (1.50×10 -3 -2.79×10 -3 ) substitutions·site -1·year -1, respectively. The effective population sizes of Cluster1 and Cluster2 tended to be stable after 2002 and 2003, respectively. In addition, Cluster1 and Cluster2 formed eleven and eight unique branches, respectively, suggesting the possibility of divergent epidemics of this genotype. A total of 35 propagation clusters were formed in the molecular propagation network, including 92 Nanjing sequences with an average degree of 4.3. Males, MSM and people with multiple sexual partners were more likely to be connected to the network. Students and young patients were more likely to be connected to the network. Conclusions:HIV-1 CRF07_BC infection was characterized by low age, multiple sexual partners, unprotected behaviors and transmission among MSM in Nanjing from 2015 to 2019. It was recommended to pay more attention to students and young people, to formulate more effective prevention and control measures for high-risk sexual behaviors, and to carry out continuous molecular monitoring of CRF07_BC strain, so as to provide scientific basis for the prevention and control of HIV CRF07_BC.

Objective:To investigate the effects of probiotics on intestinal flora, intestinal function, and T lymphocyte level in patients with cervical cancer after radiotherapy.Methods:A total of 92 patients with cervical cancer who underwent pelvic radiotherapy in The Second Affiliated Hospital of Zhengzhou University from September 2020 to February 2022 were included in this study. They were randomly divided into control and experimental groups ( n = 46/group). The patients in the experimental group took probiotics during radiotherapy, while the patients in the control group did not take probiotics during radiotherapy. The amount of intestinal flora, D-lactic acid, diamine oxidase, and T lymphocyte subset levels pre- and post-radiotherapy were compared between the two groups. Urinary lactulose (L) and mannitol (M) concentrations were determined in each group. Urinary excretion ratios of L to M were calculated. Results:After 10, 15, and 20 times of radiotherapy and after all radiotherapies, the amount of Escherichia coli and Enterococcus in the experimental group was significantly lower than that in the control group ( F = 128.60, 224.99, all P < 0.05). The amount of Bifidobacteria and Lactobacilli in the experimental group was significantly higher than that in the control group ( F = 2 065.46, 948.23, both P < 0.05). After 10, 15, and 20 times of radiotherapy and after all radiotherapies, plasma D-lactic acid level in the experimental group was (9.34 ± 1.63) μg/L, (9.15 ± 1.36) μg/L, (8.68 ± 1.06) μg/L, and (8.05 ± 0.82) μg/L, respectively. After 10, 15, and 20 times of radiotherapy and after all radiotherapies, plasma diamine oxidase level in the experimental group was (86.34 ± 20.25) μg/L, (84.28 ± 17.45) μg/L, (80.40 ± 13.35) μg/L, and (76.85 ± 10.87) μg/L, respectively, and urinary excretion ratio of L to M in the experimental group was (1.84 ± 0.16), (1.55 ± 0.12), (1.26 ± 0.09), (0.98 ± 0.06), respectively, all of which were significantly lower than those in the control group ( F = 121.60, 31.73, 417.84, all P < 0.05). After 10, 15, and 20 times of radiotherapy and after all radiotherapies, CD4 + level in the experimental group was (39.80 ± 4.90)%, (40.92 ± 5.30)%, (42.52 ± 6.14)%, (43.83 ± 6.55)%, respectively, CD4 +/CD8 + was (1.52 ± 0.25), (1.63 ± 0.22), (1.71 ± 0.39), (1.83 ± 0.22), respectively, all of which were significantly higher than those in the control group ( F = 58.69, 31.07, all P < 0.05). Conclusion:Probiotics can improve the status of intestinal flora and intestinal barrier function in patients with cervical cancer after radiotherapy, and simultaneously improve the cellular immune function of patients.

Objective    To investigate the role of preoperative peripheral blood CD4/CD8 ratio in predicting the prognosis of patients with coronary atherosclerotic heart disease (CAD) after off-pump coronary artery bypass grafting (OPCABG). Methods    A total of 118 patients with CAD who underwent OPCABG in our hospital from September 2016 to April 2017 were included in the study, including 82 males and 36 females aged 62.74±4.50 years. The primary end point was the incidence of major adverse cardiovascular events (MACE). Patients were divided into a high CD4/CD8 group (&ge;1.40, 62 patients) and a low CD4/CD8 group (<1.40, 56 patients) according to the results of flow cytometry. The correlation between CD4/CD8 ratio and prognosis of patients after OPCABG and the value of CD4/CD8 ratio for predicting postoperative MACE were evaluated. Results    Median duration of follow-up was 23.25 (20.91, 24.70) months, during which 21 patients (17.80%) experienced MACE and 4 patients (3.39%) were lost to follow-up. Kaplan-Meier analysis revealed that high CD4/CD8 group had a significantly higher MACE rate than the low CD4/CD8 group did (log-rank χ2=5.797, P=0.02). The results of adjusted Cox proportional hazards model showed that CD4/CD8 ratio (HR=3.103, 95%CI 1.557-6.187, P<0.01) was an independent risk factor of MACE in patients with CAD after OPCABG. The receiver operating characteristic curve showed that area under curve was 0.778 (95%CI 0.661-0.894, P<0.01), the optimal cut off value was 2.24, the sensitivity was 57.1%, and the specificity was 87.6%. Conclusion    Preoperative peripheral blood CD4/CD8 ratio is an independent predictor of MACE after OPCABG in patients with CAD.

The aim of this paper was to investigate the effect of Faeces Bombycis(FB) on the intestinal microflora in rats with syndrome of damp retention in middle-jiao, and to explore its mechanism in regulating intestinal microflora from the perspective of microorganisms contained in FB. The contents of antidiuretic hormone(ADH) and C-reactive protein(CRP) in serum and aquaporin 3(AQP3) in jejunum were determined by enzyme-linked immunosorbent assay(ELISA). Illumina Miseq platform was used for high-throughput sequencing of the rat feces and FB. The ELISA results showed that as compared with the normal control group, the contents of ADH and CRP in the model group were significantly increased(P<0.05), and the content of AQP3 was significantly decreased(P<0.05). After drug administration, the ADH, CRP and AQP3 contents were recovered. Sequencing of rat feces showed that the ACE, Chao1 and Shannon indexes of the intestinal microflora were the lowest in the model group. As compared with the normal control group, the levels from phylum to genus were all significantly changed in model group, and Proteobacteria, Acinetobacter, Anaerobacter, Pseudomonas, and Parabacteroides levels were significantly increased(P<0.05), while Marvinbryantia level was significantly decreased(P<0.05). As compared with the model group, Proteobacteria was significantly decreased in the FB low and high dose groups(P<0.05), and Acinetobacter, Anaerobacter, Pseudomonas, Parabacteroides levels were significantly decreased in the low, medium and high dose groups(P<0.05), while Lachnoanaerobaculum, Intestinimonas and Marvinbryantia were increased significantly in the high dose group(P<0.05). Sequencing analysis of FB showed that the relative abundance of Leclercia, Pantoea, Brachybacterium, Shimwellia, Hartmannibacter, Klebsiella, Serratia, Aurantimonas, Paenibacillus and Bacillus was high in the FB, but they were basically not present or little in the rat feces. In conclusion, FB may play a role in the treatment of "syndrome of damp retention in middle-jiao" by balancing the intestinal microflora, and this effect may be related to the metabolites of microorganisms in the FB.
Animals ; Aquaporin 3/analysis* ; Bombyx/chemistry* ; C-Reactive Protein/analysis* ; Feces/chemistry* ; Gastrointestinal Microbiome ; High-Throughput Nucleotide Sequencing ; Medicine, Chinese Traditional ; Rats ; Vasopressins/blood*

【Objective】 To investigate the feasibility of full-endoscopic posterolateral odontoidectomy through morphological analysis and cadaver specimen surgery. 【Methods】 We collected the DICOM data of 20 normal cervical CT patients （10 males and 10 females） from the PACS Image Library of our hospital. The Mimics software was used for cervical CT reconstruction and anatomical measurements were made to measure the maximum sagittal diameter， coronal diameter and height of the odontoid process. The C1 lateral mass could provide the maximum working height and width of endoscopic operation with a diameter of 7 mm， as well as the angle between the anchor point of C1 lateral mass and the notch on both sides of the odontoid process. The feasibility of endoscopic surgery was analyzed based on the measured data. The fresh frozen corpse was used for the operation in prone position under the guidance of C-arm. Kirschner wire was anchored at the midpoint of the lower surface of the C1 lateral mass. Part of the C1 lateral mass was removed by the grinding drill and endoscopic tools， and then the odontoid process and adjacent ligaments were removed. 【Results】 The maximum sagittal diameter， coronal diameter and height of the odontoid process were （11.73±0.74）mm， （10.97±0.71）mm and （14.51±0.91）mm， respectively. The working height and width of the C1 lateral mass were （13.53±0.57）mm and （10.00±1.27）mm， respectively. The angle between the anchor point and the double-edge notch of the odontoid process was （28.3±3.1）°， with no statistical difference between the male and female patients （P>0.05）. All the measurements met the requirements of 7 mm endoscopic implantation and surgical operation， and the space for swing could be provided for complete or partial removal of the odontoid process to meet the requirements of ventral spinal decompression. In cadaver surgery， a fully endoscopic posterolateral approach enabled complete removal of the odontoid process by grinding part of the C1 lateral mass. Postoperative cervical CT confirmed that the odontoid process had been completely resected， and there were no signs of dural sac or vertebral artery injury. 【Conclusion】 The odontoid process can be completely resected through a posterolateral endoscopic approach via the lateral mass approach of C1， providing a new surgical method for clinical odontoidectomy to decompress the spinal cord in craniovertebral junction.

Objective: Moxifloxacin (MFX) shows good activity against and can be a possible antibiotic therapy to treat infection; however, other studies have shown a lower or no activity. We aimed to evaluate MFX activity against using zebrafish (ZF) model . Methods: A formulation of labeled with CM-Dil was micro-injected into ZF. Survival curves were determined by recording dead ZF every day. ZF were lysed, and colony-forming units (CFUs) were enumerated. Bacteria dissemination and fluorescence intensity in ZF were analyzed. Inhibition rates of MFX and azithromycin (AZM, positive control) were determined and compared. Results: Significantly increased survival rate was observed with different AZM concentrations. However, increasing MFX concentration did not result in a significant decrease in ZF survival curve. No significant differences in bacterial burdens by CFU loads were observed between AZM and MFX groups at various concentrations. Bacterial fluorescence intensity in ZF was significantly correlated with AZM concentration. However, with increasing MFX concentration, fluorescence intensity decreased slightly when observed under fluorescence microscope. Transferring rates at various concentrations were comparable between the MFX and AZM groups, with no significant difference. Conclusion MFX showed limited efficacy against using ZF model. Its activity needs to be confirmed.
Animals ; Anti-Bacterial Agents ; pharmacology ; Disease Models, Animal ; Moxifloxacin ; pharmacology ; Mycobacterium Infections, Nontuberculous ; drug therapy ; Mycobacterium abscessus ; drug effects ; Zebrafish