OBJECTIVETo study the genotype distribution of extended-spectrum beta-lactamases (ESBLs) in ESBLs-producing Escherichia coli (E. coli) isolates from posthepatitic cirrhosis' patients with bloodstream infection.

METHODSE. coli were isolated in bloodstream from patients with posthepatitic cirrhosis between January and December in 2011. The strains were identified by VITEK-II. The antibiol susceptibility tests were performed with K-B method. beta-lactamases genes were detected multi-PCR, PCR, sequence and blast.

RESULTSA total of 79 non-duplicate clinical isolates of E coli were consecutively collected from liver cirrhosis' patients with bloodstream infection. There were 20 isolates produced TEM-1 type beta-lactamases and 1 isolate produced SHV-1 typebeta-lactamases. 40 clinical isolates were detected to produce CTX-M type ESBLs, there were 20 CTX-M-1 group and 26 CTX-M-9 group, including 6 stains habouring both CTX-M-1 and CTX-M-9 group. Eight CTX-M genotypes were confirmed by sequencing of the PCR products, including CTX-M-3, CTX-M-14, CTX-M-15, CTX-M-24, CTX-M-28, CTX-M-31, CTX-M-65 and CTX-M-79.

CONCLUSIONCTX-M genotype ESBLs was the most popular extended-spectrum beta-lactamases in E. coli isolated from liver cirrhosis' patients with bloodstream infection. The CTX-M-14 is the dominant epidemic type.

Bacteremia ; microbiology ; Cross Infection ; microbiology ; Drug Resistance, Bacterial ; Escherichia coli ; drug effects ; enzymology ; genetics ; isolation & purification ; Escherichia coli Infections ; microbiology ; Escherichia coli Proteins ; genetics ; Genotype ; Hospitalization ; statistics & numerical data ; Humans ; Liver Cirrhosis ; therapy ; Microbial Sensitivity Tests ; beta-Lactamases ; genetics ; metabolism

Classical swine fever virus (CSFV), an enveloped positive-stranded RNA virus in the genus Pestivirus of the Flaviviridae family, is the causative agent of a highly contagious swine disease characterized by symptoms of hemorrhagic fever and immune depression, usually leading to substantial economic losses. The serological methods for detection of CSFV antibody such as ELISA are important means for the diagnosis of CSFV and immune surveillance. It is difficult to obtain CSFV antigen with high quality using traditional method because its titration titer is low in cell culture. CSFV has four structural protein named C, E0, El and E2. The E2 protein contains major antigenic determinants that are conserved between different CSFV strains and involved in neutralization by antibodies. So recombinant E2 protein can be developed as an alternative to the intact viral antigen. So far, CSFV E2 have not been expressed in E. coli with high level. Many factors, such as the secondary structure, the stability of 5' and 3' terminus of gene, the location of SD sequence and the bias of codes, are involved in the expressing level of foreign gene in E. coli . In this study, two sites of the E2 gene sequence were confirmed to be detrimental to its expression efficiency in E. coli through the computer-aided analysis. So they were mutated using recombinant PCR without changing the amino acids sequence of CSFV E2 gene. A plasmid was constructed by inserting the mutated E2 gene into the prokaryotic expression vector pET-28a(+) and named pETE2. The E. coli competent host BL21 (DE3)lysS transformed with pETE2 could express the E2 gene at high level, amounting to 28% of the total protein of the induced recombinant bacteria at the presence of IPTG. Except the hydrophobic transmembrane domain at C terminus, the recombinant E2 protein includes the total aa sequence. So it contains all the potential linear antigen epitopes of E2 protein because hydrophobic aa region can not form epitope. The recombinant E2 protein was CSFV-specific as proved by Western blotting and indirect ELISA. The rabbits immunized with the recombinant E2 can be protect from the challenge of hog cholera lapinized virus. This is the first report that E2 gene is expressed with high level expression in E. coli. In conclusion, it is an effective measure that mutate the CSFV E2 gene to increase its expression level in E. coli. The recombinant CSFV E2 protein possess fine immunonicity and can be used the antigen for the detection of CSFV antibody.
Antigens, Viral ; genetics ; immunology ; metabolism ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; metabolism ; Mutagenesis, Site-Directed ; methods ; Polymerase Chain Reaction ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Viral Envelope Proteins ; genetics ; immunology ; metabolism

OBJECTIVETo study the status of beta-lactamase produced by multiresistant Aeromonas selected from cirrhosis patients to provide reference for treatment and reduce resistance and control spreading.

METHODSFour multiresistant Aeromonas strains isolated from serious liver cirrhosis patients from the No. 302 hospital. The TEM resistant genes were detected by PCR and agarose gel electrophoresis.

RESULTSThree TEM-1 positive strains were detected from four multiresistant Aeromonas isolates consisting of one Aeromonas sobria and three Aeromonas hydrophila isolated from blood and ascites. This was further confirmed by gene sequencing. The multiresistance to antibiotics was higher in four Aeromonas isolates. All strains tested were resistant to ampicillin, cefazolin and cefmetazole.The cirrhosis patients who suffered from Aeromonas infection had poor prognosis and had mortality rate of 3/4.

CONCLUSIONThe beta-lactamase TEM-1 resistant genes was detected in clinical multiresistant Aeromonas strain isolated from serious cirrhosis patients.The results suggested that TEM-1 was the main resistance mechanism of Aeromonas strain and was reduced by adding enzyme inhibitor.

Adult ; Aeromonas ; drug effects ; genetics ; isolation & purification ; Anti-Bacterial Agents ; pharmacology ; Drug Resistance, Bacterial ; genetics ; Gram-Negative Bacterial Infections ; microbiology ; Humans ; Liver Cirrhosis ; microbiology ; Male ; Middle Aged ; Prognosis ; beta-Lactamases ; genetics

Objective To study the mechanism of IL-6 stimulated the secretion of IL-10 in nasopharyngeal carcinoma CNE-2 cells.Methods Nasopharyngeal carcinoma CNE-2 cells were cultured with recombination cytokine IL-6 in vitro,or anti-IL-6 receptor antibody and signal pathway inhibitor were pre-incubated for 1 hour,and then IL-6 were added,the mRNA and protein level of IL-10 were separately detected by RT-PCR and enzyme-linked immunosorbent assay.Results Compared with the control group,the expression and secretion of IL-10 in IL-6 stimulated group were significantly increased,which was in a dose and time dependent way,the difference was significant(P ＜ 0.01).Additionally,IL-6 stimulated the expression and secretion of IL-10 by CNE-2 cells were significantly decreased in following pre-incubated with anti-IL-6 receptor antibody or NF-κB inhibitor,the difference was significant(P ＜ 0.01),but such effect was not detected when CNE-2 cells were pre-incubated with the PI3K inhibitor,p38/MAPK inhibitor,JNK inhibitor,MEK1/2 inhibitor and STAT3 inhibitor.Conclusion IL-6 can induce the expression and secretion of IL-10 in nasopharyngeal carcinoma CNE-2 cells via IL-6R/NF-κB signal pathway,and blocking IL-6 signal may be useful for the immunotherapy of nasopharyngeal carcinoma.

Drimary fetal hydrothorax(DPHT)is an uncommon congenital disease. Petal hydrothorax is thought to be due to leakage of lymphatic fluid in the pleural space either caused by direct leakage of lymphatic fluid from the thoracic duct,overproduction or impaired drainage of lymph. The outcome of fetal hydrothorax may vary from spon_taneous resolution without postnatal morbidity to severe fetal hydrops and perinatal death. At present,there is no consen_sus on the treatment of DPHT. Now,the latest researches progress on prenatal ultrasonographic characteristics,prenatal diagnostic procedures,prenatal intervention,and prognosis in DPHT,were reviewed combined with the diagnosis and treatment principle of single center in Guangdong Women and Childrenˊs Hospital,which would provide useful guidance for clinical treatment of DPHT.

Objective To investigate the clinical manifestations, diagnosis and treatment of primary intracranial tumors of multiple cellular origins. Methods The clinical data including the clinical manifestations, imaging findings, and treatment of 8 patients with primary intracranial tumors of multiple cellular origins were retrospectively analyzed, and the possible mechanism of these tumors were explored. Results A total of 16 primary intraeranial tumors were found in the 8 patients, including intraeranial meningioma (8 tumors), pituitary adenoma (3 tumors), schwannoma (2 tumors), and glioma (2 tumors). The tumors were resected in a single operation in 5 cases, and in two surgical sessions in 3 cases. Twelve tumors were radically resected, 3 were subtotally resected, and 1 was partially resected. Conclusion MR is an important diagnostic modality for primary intraeranial tumors of multiple cellular origins. Tumor resection in one or staged operations combined with radiotherapy or chemotherapy is the primary treatment option for these tumors.

Objective To evaluate the additional role of 99mTc-MIBI SPECT-CT imaging before parathyroidectomy (PTX) for secondary hyperparathyroidism (SHPT) patients.Methods A total of 359 SHPT patients from the First Affiliated Hospital of Nanjing Medical University were enrolled in the study from April 2011 to January 2016.Patients were divided into two groups according to the difference of 99mTc-MIBI imaging techniques preoperatively.Patients in group A had dual-phase 99mTc-sestamibi parathyroid scintigraphy performed only.Patients in group B had SPECT-CT parathyroid scintigraphy added in the early phase.With the parathyroid glands confirmed by pathology after operation being the referencc,scnsitivity,specificity,consistency and surgical success rates of the patients in two groups were compared.Results 359 patients (166 men and 193 women) were enrolled,among which 339 on hemodialysis and 20 on peritoneal dialysis.The average age of those patients was (45.90± 11.20) years and median dialysis age was 84 (60.0,120.0) months.Total detection sensitivity,specificity,consistency in group A (169 patients) were 65.54％,56.52％,65.24％ and in group B (190 patients) were 73.84％,84.62％,74.21％ respectively.Total sensitivity and consistency were higher in group B than those in group A (P ＜ 0.01),while no difference in total specificity was observed between two groups (P=0.055).In group A,detection sensitivity of parathyroid glands were 47.56％,77.44％,57.14％ and 79.88％ in right upper gland (RU),right lower gland (RL),left upper gland (LU) and left lower gland (LL) respectively.Consistency in group A were 48.52％,77.51％,56.21％ and 78.70％ respectively.In group B,sensitivity were 58.33％,83.78％,69.78％ and 82.89％ in RU,RL,LU and LL respectively.Consistency in group B were 59.47％,84.21％,70.53％ and 82.63％ respectively.Sensitivity and consistency of upper glands were lower than lower glands in both groups (P ＜ 0.01).When SPECT-CT was added,sensitivity and consistency of upper glands for both sides were higher in group B than those in group A,while no difference of surgical success rates was found between two groups (87.57％ vs 92.63％,P=0.107).Conclusion 99mTc-MIBI SPECT-CT can be combined with anatomic image effectively and increase the sensitivity and consistency of total and upper parathyroid glands.It can also make the accurate location of the lesions,which improves the efficiency of the operation.

Spinal muscular atrophy (SMA) is a disorder characterized by degeneration of lower motor neurons and occasionally bulbar motor neurons leading to progressive limb and trunk paralysis as well as muscular atrophy. Three types of SMA are recognized depending on the age of onset, the maximum muscular activity achieved, and survivorship: SMA1, SMA2, and SMA3. The survival of motor neuron (SMN) gene has been identified as an SMA determining gene, whereas the neuronal apoptosis inhibitory protein (NAIP) gene is considered to be a modifying factor of the severity of SMA. The main objective of this study was to analyze the deletion of SMN1 and NAIP genes in southern Chinese children with SMA. Here, polymerase chain reaction (PCR) combined with restriction fragment length polymorphism (RFLP) was performed to detect the deletion of both exon 7 and exon 8 of SMN1 and exon 5 of NAIP in 62 southern Chinese children with strongly suspected clinical symptoms of SMA. All the 32 SMA1 patients and 76% (13/17) of SMA2 patients showed homozygous deletions for exon 7 and exon 8, and all the 13 SMA3 patients showed single deletion of SMN1 exon 7 along with 24% (4/17) of SMA2 patients. Eleven out of 32 (34%) SMA1 patients showed NAIP deletion, and none of SMA2 and SMA3 patients was found to have NAIP deletion. The findings of homozygous deletions of exon 7 and/or exon 8 of SMN1 gene confirmed the diagnosis of SMA, and suggested that the deletion of SMN1 exon 7 is a major cause of SMA in southern Chinese children, and that the NAIP gene may be a modifying factor for disease severity of SMA1. The molecular diagnosis system based on PCR-RFLP analysis can conveniently be applied in the clinical testing, genetic counseling, prenatal diagnosis and preimplantation genetic diagnosis of SMA.
Child ; Child, Preschool ; China ; epidemiology ; Female ; Gene Deletion ; Genetic Predisposition to Disease ; epidemiology ; genetics ; Humans ; Incidence ; Infant ; Male ; Neuronal Apoptosis-Inhibitory Protein ; genetics ; Polymorphism, Single Nucleotide ; genetics ; Spinal Muscular Atrophies of Childhood ; epidemiology ; genetics ; Survival of Motor Neuron 1 Protein ; genetics

Objective: To investigate the effect of intrauterine intervention on severe primary fetal hydrothorax. Methods: Twelve cases with severe fetal primary hydrothorax who underwent prenatal intervention from January 2014 to December 2017 in Guangdong Women and Children Hospital were retrospectively reviewed. Results: The median gestational age of prenatal diagnosis was 30.8 weeks (24.0-33.0 weeks) . All cases were excluded congenital chromosomal abnormalities by prenatal diagnosis, and had no complications of pregnancy during prenatal diagnosis and had hydrothorax. Three cases (3/12) were right hydrothorax, the other 9 cases (9/12) were bilateral. Thoracoamniotic shunting was performed in 7 cases (7/12) . Thoracentesis was performed in 5 cases (5/12) , and the hydrothorax reappeared soon after operation in 4 cases, shunt placement was performed again. The hydrothorax was dissolved in 2 cases, released in 6 cases.Tube falling off occurred in 1 case,treatment was abandoned in 1 case and intrauterine fetal death happened in 1 case, and 1 case wasn′t rechecked by ultrasonic due to premature birth following thoracentesis. In 10 cases who had deliveries, 5 newborns (5/10) were premature, 6 newborns (6/10) underwent assisted mechanical ventilation, 8 newborns (8/10) underwent thoracic close drainage, all of them were discharged when hydrothorax resolved. Conclusions Antenatal intervention may improve the chance of survival in severe primary fetal hydrothorax. Thoracoamniotic shunting is the first-choice for the primary severe fetal hydrothorax.