Vibrio parahaemolyticus, the main pathogen causing seafood related food poisoning worldwide, has strong biofilm formation ability. ToxR is a membrane binding regulatory protein, which has regulatory effect on biofilm formation of V. parahaemolyticus, but the specific mechanism has not been reported. c-di-GMP is an important second messenger in bacteria and is involved in regulating a variety of bacterial behaviors including biofilm formation. In this study, we investigated the regulation of ToxR on c-di-GMP metabolism in V. parahaemolyticus. Intracellular c-di-GMP in the wild type (WT) and toxR mutant (ΔtoxR) strains were extracted by ultrasonication, and the concentrations of c-di-GMP were then determined by enzyme linked immunosorbent assay (ELISA). Three c-di-GMP metabolism-related genes scrA, scrG and vpa0198 were selected as the target genes. Quantitative real-time PCR (q-PCR) was employed to calculate the transcriptional variation of each target gene between WT and ΔtoxR strains. The regulatory DNA region of each target gene was cloned into the pHR309 plasmid harboring a promoterless lacZ gene. The recombinant plasmid was subsequently transferred into WT and ΔtoxR strains to detect the β-galactosidase activity in the cellular extracts. The recombinant lacZ plasmid containing each of the target gene was also transferred into E. coli 100λpir strain harboring the pBAD33 plasmid or the recombinant pBAD33-toxR to test whether ToxR could regulate the expression of the target gene in a heterologous host. The regulatory DNA region of each target gene was amplified by PCR, and the over-expressed His-ToxR was purified. The electrophoretic mobility shift assay (EMSA) was applied to verify whether His-ToxR directly bound to the target promoter region. ELISA results showed that the intracellular c-di-GMP level significantly enhanced in ΔtoxR strain relative to that in WT strain, suggesting that ToxR inhibited the production of c-di-GMP in V. parahaemolyticus. qPCR results showed that the mRNA levels of scrA, scrG and vpa0198 significantly increased in ΔtoxR strain relative to those in WT strain, suggesting that ToxR repressed the transcription of scrA, scrG and vpa0198. lacZ fusion assay showed that ToxR was able to repress the promoter activities of scrA, scrG and vpa0198 in both V. parahaemolyticus and E. coli 100λpir. EMSA results showed that His-ToxR was able to bind to the regulatory DNA regions of scrA and scrG, but not to the regulatory DNA region of vpa0198. In conclusion, ToxR inhibited the production of c-di-GMP in V. parahaemolyticus via directly regulating the transcription of enzyme genes associated with c-di-GMP metabolism, which would be beneficial for V. parahaemolyticus to precisely control bacterial behaviors including biofilm formation.
Vibrio parahaemolyticus/metabolism* ; Escherichia coli/metabolism* ; Bacterial Proteins/metabolism* ; Transcription Factors/genetics* ; Gene Expression Regulation, Bacterial

OBJECTIVE: To detect fusion gene with pathological significance in a patient with refractory and relapsed acute B cell lymphoblastic leukemia (B-ALL) and to explore its laboratory and clinical characteristics. METHODS: Transcriptome sequencing was used to detect potential fusion transcripts. Other laboratory results and clinical data of the patient were also analyzed. RESULTS: The patient was found to harbor TCF3 exon 17-ZNF384 exon 7 in-frame fusion transcript. The minimal residual disease (MRD) has remained positive after multiple chemotherapy protocols including CD19-, CD22- targeted chimeric antigen receptor T cells immunotherapy. The patient eventually achieved complete remission and sustained MRD negativity after allogeneic hemopoietic stem cell transplantation (allo-HSCT). CONCLUSION Transcriptome sequencing can effectively detect potential fusion genes with clinical significance in leukemia. TCF3-ZNF384 positive B-ALL has unique laboratory and clinical characteristics, may not well respond to chemotherapy and immunotherapy, and is more likely to relapse. Timely allo-HSCT treatment may help such patients to achieve long-term disease-free survival. TCF3-ZNF384 positive B-ALL is not uncommon in pediatric patients but has not been effectively identified.
B-Lymphocytes ; Basic Helix-Loop-Helix Transcription Factors/genetics* ; Child ; Hematopoietic Stem Cell Transplantation ; Humans ; Laboratories ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/therapy* ; Trans-Activators/genetics* ; Transcriptome

Radiation therapy plays an important role in the adjuvant treatment of patients with early endometrial carcinoma. Vaginal stump is a common site of disease failure for early endometrial carcinoma patients with intermediate-high risk factors for recurrence. Compared with external beam radiotherapy, vaginal brachytherapy (VBT) can achieve comparable local control rate with fewer toxicities. In this article, research progresses upon the application of VBT in patients with early endometrial carcinoma after hysterectomy were investigated from multiple perspectives of the selection of patients, the selection of vaginal applicator, factors influencing dose distribution, image-guided adaptive brachytherapy, the design and implementation of radiotherapy regime. In addition, the application of intensity-modulated VBT and the usage of novel quality assurance equipment were also discussed.

Objective:To study the transcriptional regulation of pilABCD by the master quorum sensing (QS) regulator OpaR in Vibrio parahaemolyticus. Methods:Total RNAs were extracted from the wild type (WT) and opaR mutant (Δ opaR) strain. Quantitative real-time PCR (qPCR) was employed to calculate the transcriptional variation of pilA (the first gene of pilABCD operon) between WT and Δ opaR. The regulatory DNA region of pilABCD was cloned into the corresponding restriction endonuclease sites of pHRP309 harboring a promoterless lacZ reporter gene. The recombinant pHRP309 plasmid was then transferred into WT and Δ opaR, respectively, to detect the β-galactosidase activity in cellular extracts using a β-Galactosidase Enzyme Assay System (Promega). The primer extension assay was applied to map the transcription start site of pilABCD using the total RNAs extracted from the WT strain as the template. The regulatory DNA region of pilABCD was amplified by PCR, and the over-expressed His-OpaR was purified under native conditions with nickel loaded HiTrap Chelating Sepharose columns (Amersham). Thereafter, the electrophoretic mobility shift assay (EMSA) was applied to analyze the DNA-binding activity of His-OpaR to the target DNA in vitro, and the DNase I footprinting assay was further employed to detect the DNA-binding sites of His-OpaR within the target DNA. Results:The results of qPCR and LacZ fusion assays showed that OpaR activated the transcription of pilABCD, leading to a gradual increase in the expression level of pilA with the extension of culture time. The primer extension assay detected only one transcription start site located at 155 bp upstream of pilA. The results of EMSA and DNase Ⅰ footprinting assays showed that His-OpaR protected two DNA regions located from -246 to -197 bp and -181 to -131 bp upstream of pilA. Conclusions:Vibrio parahaemolyticus OpaR activated the transcription of pilABCD in a direct manner.

Through the effort in nearly 40 years of Tanzania, the total number of reported cases, the prevalence of HIV/AIDS and the number of death have declined, while the number of patients receiving antiretroviral therapy has increased significantly in Tanzania. At the same time, however, there are new challenges that require more attention, such as the HIV spread to rural areas, middle and lower social classes, and female teenagers. Although the overall performance of the HIV/AIDS prevention and control response is great, there is still a need to survey the new trends of this disease at the micro-level in Tanzania. In addition, there is a necessity of interdisciplinary concern due to the prevalence of heterosexual anal sex in Tanzania.

OBJECTIVE: To study whether negative-pressure septal drainage could be an alternative to packs after septoplasty. METHOD: This was a randomized controlled trial. The study involved 60 patients who underwent septoplasty. Patients were randomly divided into two groups, one with anterior nasal packs and the other with negative-pressure septal drainage. Patients were asked to record pain levels using a visual analogue scale (VAS). Postoperative symptoms and complications were compared during 24 h and 48 h postoperative period including pain, drying sensation of mouth, sleep difficulty, conjunctival congestion, haemorrhage. VAS scores and incidence were evaluated during 1 week and 6 weeks postoperative period including pain, bleeding, haematoma, septal perforation, synechiae and septal perforation. RESULT: Patients of negative-pressure septal drainage suffered from less pain than patients of nasal packs during the first 24 h and 48 h postoperative period. The results for pain, drying sensation of mouth, sleep difficulty, conjunctival congestion, haemorrhage were different between groups (P < 0.05), especially the amount of bleeding during 48 h postoperatively in patients undergoing negative pressure drainage [(0.52 ± 0.63)ml] was significantly less than the group who received anterior nasal packs [(21.03 ± 5.88) ml] (P < 0.01). On the other hand, haematoma, synechiae and perforation were not statistically different between groups during 1 week and 6 weeks follow-up period (P > 0.05). CONCLUSION Using negative-pressure drainage instead of nasal packs after septoplasty seems a more reasonable option. The negative-pressure drainage technique may be the preferred option to provide higher patient satisfaction and has the same level of postoperative complica.tion to nasal packs as for septoplasty surgery.
Drainage ; Humans ; Nasal Septum ; surgery ; Nasal Surgical Procedures ; Negative-Pressure Wound Therapy ; methods ; Nose ; Pain Measurement ; Patient Satisfaction ; Postoperative Period ; Tampons, Surgical

Objective To investigate the expression of IL-33,IL-37 in nasal polyps tissue,and their relevance to the pathogenesis of human nasal polyps. Methods Immunohistochemistry was used to detect the expression of IL-33,IL-37, in excisional nasal polyps tissue from 40 patients with nasal polyps. In addition,the expression of IL-33,IL-37 was also detected in normal nasal mucosa from 18 patients with the nasal septum deviation. Results In the nasal polyps tissues,IL-33 positive rate was 65.0%,IL-37 positive rate was 20.0%.In the normal nasal mucosa, IL-33 positive rate was 33.3%,IL-37 positive rate was 77.8%. Conclusion IL-33 and IL-37 play an important role in the pathogenesis of nasal polyps.

BACKGROUNDHearing impairment has been reported to be common in patients with mitochondrial disorders, a group of diseases characterized by pleiomorphic clinical manifestations due to defects in oxidative phosphorylation of mitochondria. This study aimed to investigate the audiological characteristics in a large cohort of patients with mitochondrial disease.

METHODSComprehensive audiological evaluations, including pure tone audiometry, tympanometry, speech audiometry, otoacoustic emissions, electrocochleography and auditory brainstem evoked potentials, were performed in 73 Chinese patients with mitochondrial encephalomyopathy and with confirmed mitochondrial DNA (mtDNA) defects.

RESULTSAmong the patients, 71% had hearing impairment. However, the incidence rate and severity of hearing impairment were much less in the chronic progressive external ophthalmoplegia (CPEO) subtype than in the mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS), myoclonic epilepsy with ragged red fibers (MERRF) and Kearns-Sayre syndrome (KSS) subtypes. While most of our patients had a predominantly cochlea origin for the hearing deficit, five patients had an auditory neuropathy spectrum disorder and three patients had impairment of both cochlea and auditory cortex.

CONCLUSIONSVarious portions of the auditory system could be involved in patients with mitochondrial diseases, including cochlea, auditory nerve, auditory pathway and cortex. Hearing loss was more associated with multisystem involvement. Genotype, mutant load of mtDNA and other unknown factors could contribute to heterogeneity of hearing impairment in mitochondrial disease.

Adolescent ; Adult ; Aged ; Child ; Female ; Hearing Loss ; physiopathology ; Hearing Loss, Central ; physiopathology ; Humans ; Male ; Middle Aged ; Mitochondrial Encephalomyopathies ; physiopathology ; Young Adult

Objective To study the characteristics of auditory brainstem responses with different forward - masking intervals in newborns with normal hearing ,and to find out the most efficient interval .Methods Auditory evoked brainstem responses were recorded from 30 newborns(60 ears) ,with forward - masking intervals of 4 ms ,8 ms ,16 ms ,32 ms ,and unmasked as control .The ABR data were statistically analyzed with SPSS 13 .0 .Results With forward - masking intervals ,all waves Ⅰ ,Ⅲ ,Ⅴ latencies and Ⅰ - Ⅴ ,Ⅰ - Ⅲ ,Ⅲ - Ⅴ interpeak latencies were significantly (P< 0 .01) prolonged compared with those of in the unmasked ,except wave Ⅲ and Ⅰ - Ⅲ interpeak latencies with 4 ms forward - masking intervals .The waves from 8 ms interval were the most prolonged .Conclusion Forward masking increases human newborn ABR latencies and interpeak latencies .The interval time influenced the effect ,and 8 ms interval is the most efficient .

Objective To estimate the degree of oxidative stress and atherosclerosis praecox in patients with systemic lupus erythematosus (SLE), and emphasize on exploring AOPPs' clinical significance in premature atherosclerosis in SLE. Methods The levels of AOPPs, Hey, MDA, SOD, baPMV and CIMT were detected by ELISA and spectropholometry in 44 non-menopausal female SLE patients and 31 healthy middle-aged women respectively, and baPWV. The results were compared with AOPPs of the two groups. Then each group was stratified based on disease duration (≥5 years or <5 years) and the disease activity(active and inactive) in SLE patients. The patients' TC, TG, LDL were analyzed. T test, t' test and Pearson correla-tion were selected. Results The levels of AOPPs, Hcy, MDA in SLE patients were significantly higher than those of the healthy controls (P<0.05). The activities of SOD were lower than controls (P<0.05). The levels of AOPPs, Hcy, MDA, SOD had statistical significance between SLE patients disease duration ≥5 years or <5 years, active or inactive groups. There were two cases with CAP in patients with SLE (more than 5 years disease duration),while there wasnone in healthy controls. The levels of baPWV and CIMT in patients with SLE were higher than healthy controls (P<0.05), and had statistical significance in SLE patients disease duration more than or less than 5 years (P<0.05). The oxidative stress targets (AOPPs, Hey, MDA, SOD) had significant correlation with the level of baPWV and CIMT (P<0.01). The level of serum AOPPs had significant positive correlation with the levels of Hcy, TC, TG and LDL (P<0.01～0.05). Conclusion SLE patients have increased oxidative stress , and significantly higher prevalence of atherosclerosis than healthy controls. The disease duration and oxidative stress play important roles in the duration of atherosclerosis. AOPPs probably involves in the accelerated atherosclerosis of SLE patients. It may be a predictor for SLE complicated with atherosclerosis praecox.