OBJECTIVES: To detect prfA and hly toxin genes of Listeria monocytogenes using polymerase chain reaction (PCR) and colloidal gold technology. METHODS: L. monocytogenes DNA was extracted by boiling method. With prfA and hly of L. monocytogenes as the target genes, the 5' ends of upstream and downstream primers of prfA gene were labeled with 6-FAM and biotin, and the 5' ends of upstream and downstream primers of hly gene were labeled with digoxin and biotin, respectively, to establish the toxin gene detection method. Using cloning transformation, sequencing analysis, cloning of positive control products, the detection kid was developed and its specificity, sensitivity, reproducibility and stability were tested, followed by verification with sample testing. RESULTS: The concentration of L. monocytogenes DNA extracted by boiling method was 148.81±0.97 ng/μL, and the A₂₆₀/A₂₈₀ ratio ranged from 1.8 to 2.0. The PCR products showed a 100% homology with the gene sequences in GenBank database after cloning, transformation and sequencing. The colloidal gold strip yielded positive results only for L. monocytogenes samples without cross-reactions with Staphylococcus aureus, Escherichia coli or Bacillus cereus, and its minimum detection limit was 10^-2 ng/μL, demonstrating a 10-fold greater sensitivity of the test than agarose gel electrophoresis. The test also showed good reproducibility of the results when performed by different operators with good stability of the test strips after storage for 6 to 12 months. The test results showed that this kit could accurately and quickly detect L.monocytogenes in the test samples. CONCLUSIONS The detection kit developed in this study can simultaneously detect prfA and hly toxin genes of L. monocytogenes with good specificity, sensitivity, reproducibility and stability for use in food safety inspection.
Listeria monocytogenes/isolation & purification* ; Gold Colloid ; Bacterial Toxins/genetics* ; Polymerase Chain Reaction/methods* ; Hemolysin Proteins/genetics* ; Bacterial Proteins/genetics* ; DNA, Bacterial/genetics* ; Food Microbiology ; Heat-Shock Proteins

Medical microbiology experiment is faced with many problems in online teaching. This study adopts the teaching mode of online live broadcast + operation video + virtual experiment, and make up the operation gap to some extent through operation video and virtual experiment. The mode of assessment is subjective thinking question (closely following the operation process) + experiment design + literature review (focusing on the key technology or new technology of clinical assessment that cannot be carried out due to the limitation of conditions in traditional experiments, such as mass spectrometry, fluorescence quantitative PCR, and G-test), and it is helpful to understand students' mastery of teaching objectives, and the ability of comprehensive application and innovative thinking. The student questionnaire shows that most students hold a positive attitude towards the online experimental teaching mode, and the quality of students' homework shows that most students have a good learning effect.

Peptides that are composed of dextrorotary (d)-amino acids have gained increasing attention as a potential therapeutic class. However, our understanding of the in vivo fate of d-peptides is limited. This highlights the need for whole-body, quantitative tracking of d-peptides to better understand how they interact with the living body. Here, we used mouse models to track the movement of a programmed death-ligand 1 (PD-L1)-targeting d-dodecapeptide antagonist (DPA) using positron emission tomography (PET). More specifically, we profiled the metabolic routes of [⁶⁴Cu]DPA and investigated the tumor engagement of [⁶⁴Cu/⁶⁸Ga]DPA in mouse models. Our results revealed that intact [⁶⁴Cu/⁶⁸Ga]DPA was primarily eliminated by the kidneys and had a notable accumulation in tumors. Moreover, a single dose of [⁶⁴Cu]DPA effectively delayed tumor growth and improved the survival of mice. Collectively, these results not only deepen our knowledge of the in vivo fate of d-peptides, but also underscore the utility of d-peptides as radiopharmaceuticals.

Objective:To investigate the effect of arthroscopically-assisted open reduction and internal fixation of intra-articular distal radius fracture.Methods:A retrospective cohort study was made on clinical data of 44 patients with distal radial intraarticular fracture admitted to Jing′an District Central Hospital, Fudan University between June 2017 and August 2020. There were 13 males and 31 females, at age of 35-85years [(62.5±12.9)years]. According to AO/OTA fracture classification system, there were 7 patients with type B and 37 with type C. Open reduction and internal fixation with volar plate was used in all patients, among which 22 were operated on using arthroscopy assistance (arthroscopy group) and 22 were operated on with traditional intraoperative fluoroscopy (fluoroscopy group). The operation time in both groups and triangular fibrocartilage complex (TFCC) injury and fracture displacement in arthroscopy group were recorded. Patient-rated wrist evaluation (PRWE) score, disabilities of the arm, shoulder and hand (DASH) questionnaire and range of wrist motion were compared between the two groups at 12 months after operation. The incidence of complications was observed.Results:All patients were followed up for 12-15 months [(13.3±1.1)months]. The operation time in arthroscopy group was (104.0±40.5)minutes, longer than (71.3±32.1)minutes in fluoroscopy group ( P<0.05). In arthroscopy group, 14 patients (64%) with TFCC injury were diagnosed intraoperatively, with the fracture displacement gap and step for 0.8 (0.3, 0.8)mm and 1.0 (0.3, 1.5)mm under arthroscopic vision, which were reduced to 0.3 (0.0, 0.5)mm and 0.5 (0.0, 0.5)mm after arthroscopically-assisted reduction (all P<0.05). The PRWE score in arthroscopy group was (9.8±4.9)points at 12 months after operation, lower than (13.4±5.8)points in fluoroscopy group ( P<0.05). The DASH questionnaire in arthroscopy group was (9.0±5.0)points at 12 months after operation, lower than (13.0±6.1)points in fluoroscopy group ( P<0.05). The dorsal extension and posterior rotation of the wrist in arthroscopy group were (73.8±8.9)° and (82.5±8.0)°, higher than (65.8±14.2)° and (76.3±10.4)° in fluoroscopy group (all P<0.05). There were no postoperative complications such as loosened or broken screws, vascular nerve damage, incision infection or traumatic arthritis in both groups. Conclusion:Arthroscopic-assisted open reduction and internal fixation of intra-articular distal radius fracture can increase the accuracy of joint surface reduction, improve postoperative wrist function and confirm the diagnosis of TFCC injury during operation.

Objective: To study the efficacy and safety of sofosbuvir and daclatasvir regimens for patients who received kidney transplantation (KT) with hepatitis C virus (HCV) infection. Methods: This study enrolled a prospective cohort of consecutive KT patients with HCV infection from March 2016 to January 2018 in the hepatology Department of the Second Hospital of Shandong University. They were given sofosbuvir combined with daclatasvir, with or without ribavirin. The course of treatment was 12 weeks or 24 weeks. Clinical assessment, conventional liver and kidney biochemical parameters, hemoglobin, serum HCV RNA, as well as the types of immunosuppressive drugs and their doses were assessed routinely as follows: at the beginning of treatment; 2, 4, and 8 wk post treatment; at the end of treatment (EOT); and at 12, 24 wk after the therapy was completed. Adverse events and adjustment of anti-rejection drugs were surveilled during the treatment period. Results: A total of 13 patients were enrolled. All patients were naive to treatment. Their mean age was 46.84±7.79 years. There were 10 males and 3 females, 3 patients had cirrhosis (1 cases had decompensated cirrhosis), 10 patients had no cirrhosis. They were infected with HCV genotype 1 (6/13 GT1b), genotype 3 (2/13 GT3a) and genotype 6 (3/13 GT6a), and genotype 2 (2/13 GT2a). Twelve patients′ estimated glomerular filtration rate (eGFR) was > 30 ml/min per 1.73 m² at the beginning of treatment, 1 patient′s eGFR was <30 ml/min·1.73 m^2; 9 patients received 12 wk therapy, 4 patients received 24 wk therapy. Twelve patients had undetectable viral load by week 4 of treatment. All patients had undetectable HCV viral load at the end of treatment. Sustained virological response (SVR) 12 rate was achieved in 100% (13/13) of the recipients. The basic renal function remained stable during the course of treatment. No serious adverse events were observed during the treatment. Antiviral therapy was not discontinued due to side effects in any patient. Conclusions Sofosbuvir and daclatasvir for treatment of KT patients with HCV infection are highly effective and safe.

Objective Using of cumulative live birth rate(CLBR)per oocytes retrieved cycle,to assess the clinical outcomes of in vitro fertilization or intracytoplasmic sperm injection(IVF/ICSI),and to explore impact factors on CLBR following utilization of all fresh and frozen embryos in one complete IVF/ICSI cycle using gonadotropin-releasing hormone(GnRH)agonist, GnRH-antagonist and clomiphene mild stimulation protocols. Methods Of the patients who underwent IVF/ICSI from January 1st, 2014 to December 31st, 2015 in the First Affiliated Hospital, Nanjing Medical University, a total of 6 142 oocytes retrieved cycles were included. The clinical and laboratory parameters of different ovarian stimulation protocols, and the effects of the age, number of oocytes retrieved and number of embryos available on the CLBR of each oocytes retrieved cycle were analyzed.Results The CLBR was 69.0%(2 004/2 906)in the GnRH-agonist protocol versus 67.4%(644/955)in the GnRH-antagonist protocol (P>0.05); the CLBR of clomiphene mild stimulation protocol was 53.2%(1 215/2 281),significantly lower than those of the other two protocols (all P<0.05). The CLBR significantly decreased with age increased. When divided into four groups according to the patients′ age, we found that CLBR were not statistically significant using three different protocols in the 20-25 years old group(all P>0.05).There was a strong association between the number of oocytes retrieved and embryos available on CLBR. CLBR rose significantly with an increasing number of oocytes up to 6, then the rising trend slowed down. Patients were categorized into four groups according to the number of oocytes retrieved,CLBR was significantly higher using GnRH-antagonist protocol (50.0%)than mild stimulation protocol(37.0%)in low ovarian responder(0-4 oocytes)group(P<0.05). The CLBR were no significant difference among three protocols in normal(10-15 oocytes)and high responders(≥15 oocytes)group(all P>0.05).The incidence rate of ovarian hyperstimulation syndrome in GnRH-agonist protocols(5.2%,152/2 906)were significantly higher than those of GnRH-antagonist(4.4%, 42/955)and clomiphene mild stimulation protocols(1.5%,34/2 281;all P<0.05).Conclusions CLBR is an important index to assess the clinical outcomes of IVF/ICSI. Age, number of oocytes retrieved and embryos available could affect CLBR obviously. According to the different age and ovarian response of patients, we should design ovarian stimulation protocols based on target oocytes number in order to get higher CLBR and reduce complications.

Objective To study the effect of microRNA-320 (miR-320) targeting E2F1 gene on tumor glycometabolism in colorectal cancer.Methods The miR-320 expression level in colorectal cancer cell lines and cancer tissues was detected using quantitative real-time polymerase chain reaction (qRT-PCR).The binding sites of miR-320 and E2F1 were predicted by bioinformatics.Luciferase assay was used to detect the targeting regulation of miR-320 on E2F1.The relationship between E2F1 and miR-320 was verified in mRNA level and protein level.When the miR-320 in SW480 and LOVO ceils was up-regulated and the E2F1 was down-regulated,the changes of glycometabolism in tumor cells were analyzed using glucose/glucose oxidase kit and lactate test kit.Results The qRT-PCR results showed low expressions of miR-320 in colorectal cancer cell lines and cancer tissues (F =42.327,P ＜ 0.001;t =4.345,P =0.023).Luciferase assay showed that miR-320 could negatively regulate the expression of E2F1 (t =4.716,P =0.042).The expression levels of E2F1 protein and mRNA (t =4.780,P =0.041;t =5.506,P =0.031) confirmed that miR-320 could interact with E2F1 in LOVO and SW480 cells.Overexpression of miR-320 could reduce the contents of glucose (t =5.262,P=0.034;t =21.079,P=0.002) and lactic acid (t =9.609,P=0.011;t =18.582,P=0.003) in the cellular supematant in SW480 and LOVO ceils.Down-regulating the expression of E2F1 at the same time could enhance the inhibitory effect of miR-320 on glucose (t =5.128,P =0.036;t =5.089,P =0.037) and lactic acid (t =8.573,P =0.013;t =13.364,P =0.006).Conclusion E2F1 is the target gene of miR-320,and miR-320 can regulate the glycometabolism of colorectal cancer cells by targeting E2F1 gene.

Objective To investigate the risk factors of standard selectivity biliary cannulation in endoscopic retrograde cholangiopancreatography (ERCP) for patints with choledocholithiasis,and the effect and safety of assistant cannulation methods.Methods The clinical data were retrospectively analyzed in 372 patients,who received ERCP for choledocholithiasis in the Yijishan Hospital of Wannan Medical College from January 2014 to December 2014.The risk factors of standard biliary cannulation were assessed by univariate and multivariate logistic regression analysis.The success rate of assistant biliary cannulation and the incidence of post-ERCP pancreatitis (PEP) of standard biliary cannulation were compared with doubleguide wire technique (DGT) and precut sphincterotomy (PS),which were used when standard biliary cannulation failed.Results The univariate analysis showed that floppy and long duodenal papilla (P=0.000),angulated common bile duct(P=0.013) were related to failure of standard biliary cannulation.Multivariate analysis showed that floppy and long papilla (P=0.000,OR=0.131,95％CI:0.056-0.307),angulated common bile duct (P =0.003,OR =0.378,95 ％ CI:0.197-0.726) were independent risk factors for standard biliary cannulation.The success rate of standard biliary cannulation was 83.3％ (310/372),62 cases undergone DGT or PS after standard biliary cannulation failure,and the total success rate of cannulation was 99.7％ (371/372).There were 29 cases suffered from PEP in 371 successful cases of biliary cannulation,including 18 cases (5.8％,18/310) of standard biliary cannulation,8 cases (16.3％,8/49) of DGT,and 3 cases (25.0％,3/12) of PS.The incidence of PEP by DGT (x2 =5.532,P =0.019) and PS (x2 =6.994,P=0.008) was significantly higher than that of standard biliary cannulation,but there was no statistical difference on the incidence of PEP between DGT and PS (x2 =0.079,P =0.778).Conclusion Floppy and long duodenal papilla and angulated common bile duct could easily induce the failure of standard biliary cannulation in ERCP for patients with choledocholithiasis.DGT and PS are effective assistant cannulation methods when the standard cannulation fails.

Objective To study the alterations of renal function in patients with acute ischemic stroke and diabetes and to analyze the risk factors of contrast-induced nephropathy (CIN) after digital subtraction angiography (DSA).Methods Eight hundred and seventy-one cerebral infarction patients with diabetes who underwent DSA were selected in the Third Affiliated Hospital of Chongqing Medical University and Nanjing Brain Hospital Affiliated to Nanjing Medical University from August 2012 to August 2016.The patients were divided into diabetic group (n =178) and non-diabetic group (n =693).The alterations of renal function and the incidence rate of CIN were observed between two groups 3 days after DSA.Univariate analysis and multi-factor Logistic regression analysis were used to analyze the independent risk factors of CIN.Results The levels of estimated glomerular filtration rate (eGFR,ml · min-1 · 1.73 m-2) in diabetic group at 1,2 and 3 days after DSA(81.94 ±9.38,75.36 ±8.21,84.43 ±9.72) were lower than that in non-diabetic group (84.62 ± 10.06,79.08 ±9.84,87.62 ± 10.15,t =3.213,4.645,3.772,all P ＜ 0.05).The levels of serum creatinine (Scr) and cystatin (CysC) in diabetic group at 1,2 and 3 days after DSA (Scr:85.63 ±9.83,92.37 ±10.07,83.43 ±9.07;CysC:1.08 ±0.12,1.35 ±0.14,0.95 ±0.10) were higher than that in non-diabetic group (Scr:81.36 ± 8.98,87.84 ± 9.85,80.31 ± 8.64,t =5.548,5.448,4.253;CysC:0.97 ±0.11,1.21 ±0.12,0.88 ±0.09;t =11.677,13.400,9.043;all P ＜ 0.05).The incidence rate of CIN in diabetic group (25.84％ (46/178)) was higher than that in nondiabetic group (7.07％ (49/693),x2 =51.358,P =0.001).Multi-factor Logistic regression analysis showed allergies,plasma brain natriuretic peptide,heart failure,the original renal insufficiency,NIHSS score,contrast agent dosage,preoperative eGFR,preoperative Scr and preoperative CysC were the independent risk factors of CIN in cerebral infarction patients with diabetes.Conclusions The renal function decreased significantly and the incidence rate of CIN was high in cerebral infarction patients with diabetes after DSA.Allergies,plasma brain natriuretic peptide,heart failure,the original renal insufficiency,contrast agent dosage,preoperative eGFR,preoperative Scr and preoperative CysC were the independent risk factors of CIN in cerebral infarction patients with diabetes.

OBJECTIVETo explore the clinical effects of negative pressure wound therapy (NPWT) combined with artificial dermis grafting and autologous skin grafting on repair of open joint wounds and/or wounds with exposed bone fracture.

METHODSEleven patients with open joint wounds and/or wounds with exposed bone fracture, hospitalized from November 2008 to November 2014, were enrolled in the study. According to the differences of the first stage treatment, all patients were divided into experimental group ( n = 6, including 4 patients of open joint wounds, 1 patient of wound with exposed bone fracture, and 1 patient of open joint wound with exposed bone fracture), and control group ( n 5, including 2 patients of open joint wounds, 2 patients of wounds with exposed bone fracture, and 1 patient of open joint wound with exposed bone fracture). After debridement, the wounds in both groups were grafted with punctured artificial dermis, while NPWT was only used over the artificial dermis of experiment group for 1 week. In the operation at sacsod stage, autologous split-thickness skin was grafted on the vascularized artificial dermis in both groups. Results In 5 patients of open joint wounds in experimental group, the artificial dermis was vascularized well, autologous skin grafts survived, and wounds were healed. In 3 patients of open joint wounds in control group, the artificial dermis grafting all failed due to local infection, and then these wounds were repaired with local tissue flap grafting. Artificial dermis in 3 patients of wounds with exposed bone fracture in both groups was vascularized well after grafting, and the wounds were healed after autologous skin grafting, whether or not NPWT was used.

CONCLUSIONSNPWT combined with artificial dermis grafting and autolognus skin grafting can be used for repairing open joint wounds and/or wounds with exposed bone fracture.

Debridement ; Dermis ; transplantation ; Humans ; Negative-Pressure Wound Therapy ; methods ; Skin Transplantation ; methods ; Skin, Artificial ; Surgical Flaps ; Wound Healing