To obtain the tervalent fusion toxin gene (named FT),three toxin gene fragments from three species of Vibrio parahaemolyticus,Vibrio vulnificus and Vibrio mimicus were connected with the flexible linker (GGGGS) using overla Pextension PCR. The three toxin gene fragments respectively encode the mature proteins of the thermostable direct hemolysin (TDH) of V. parahaemolyticus,the cytotoxin (VVC) of V. vulnificus and the heat-labile hemolysin (VMH) of V. mimicus. The identity of FT nucleic acid sequence was 99.6% with the corresponding toxin gene fragments. The open reading frame of FT was 3225 bp,encoding 1074 amino acid residues with the predicted molecular weight (MW) of 120.4 kDa. Then,FT was subcloned into the expression vector pET-22b(+). The construction of recombinant expression vector pET-22b-FT was followed by transforming into E. coli BL21(DE3) for expression. The SDS-PAGE electrophoresis results indicated that the MW of the fusion toxin protein was matched to the predicted MW. After induction by 1 mmol/L IPTG at 37℃,the fusion toxin protein was effectively expressed in E. coli BL21(DE3) with the amount of 11.49% through thin layer chromatography scanning (TLCS) analysis. Cavia cobaya was immunized using the purified cytorrhyctes to produce the anti-serum. Through the determination of the optimum working conditions,the sensitivity test,the specificity test,repeatability test and sample simulation test,the indirect ELISA method was established,which is a broad-spectrum,rapid and specific to detect various of food-poisoning Vibrio simultaneously.

Objective To retrospectively study the biodistribution of 68Ga-DOTA-TATE as a SSTR imaging agent in human subjects.Methods A total of 106 patients with suspected disease were enrolled in this study.All patients were histologically proven for having either a single tumor ＜2 cm or without evidence of tumor during follow-up.Patients underwent PET/CT whole-body scan 17-100 min after intravenous injection of 55.2-220.0 MBq 68Ga-DOTA-TATE.ROI was drawn for measuring SUV of tracer-avid pathologies.One-way analysis of variance and two-sample t test were used for statistical analysis.Results High 68Ga-DOTA-TATE avidity was found in pituitary,with SUVmax of 4.00± 1.21.Tracer was excreted mainly through urinary system resulting in highest uptake in the urinary tracts.The SUVmax of kidney cortex was 19.01 ± 5.45.Mediastinal blood pool and liver SUVmax were 0.93±0.33 and 7.69±2.26,respectively.Mild uptake of 68Ga-DOTA-TATE was found in the brain,cerebellum,lung and muscle,all lower than that of mediastinal blood pool.Moderate accumulation of 68 Ga-DOTA-TATE (close to or slightly higher than liver) was found in adrenal gland and spleen,with SUVmax 7.61 ± 3.42 and 8.63± 2.31,respectively.Other organs (pituitary,salivary gland,thyroid,pancreas,small intestine,colon,uterus,prostate and bone) showed tracer uptake in the range between those of mediastinal blood pool and liver.68Ga-DOTA-TATE distribution in pancreas was not uniform.Nine patients had focal accumulation in the uncinate process of pancreas with highest SUVm~ up to 8.48.However,the SUVmax and SUV in the rest of pancreas (head,neck,body and tail) showed insignificant difference (F values:0.703,0.563,both P＞0.05).68Ga-DOTA-TATE uptake in each organ reached equilibrium quickly after injection but with slight increase over time.The changes in SUV,however,showed insignificant difference among organs,including different parts of pancreas (t values:from -0.09 to 1.75,from-1.70 to-0.42,respectively,all P＞0.05).Conclusions The biodistribution of 68Ga-DOTA-TATE reaches equilibrium shortly after intravenous administration and is stably maintained.The biodistribution activities are organ-specific,and characteristic to that of SSTR concentration.

OBJECTIVETo investigate the constitution and health status of the Beijing residents, and the primary influencing factors on nutrition and health, as to finding out the epidemiological characters and the changing trend of chronic diseases and to establish scientific basis for the establishment of public health and disease prevention policies of Beijing.

METHODSA stratified multi-stage cluster randomly sampling method was used. In total, 18 districts were surveyed, and among them, 9 were conducted dietary survey. The investigation was based on household, and all family members were surveyed at their home. After having got the compliance from the family members, question asking, medical examination, laboratory test and dietary investigation were carried out in these people.

RESULTSThe incidence rate of hypertension was 25.0% for the resident > or = 15 year-old, and over 50% for middle-aged and elderly people (> or = 45 years old). The incidence of diabetes for those not younger than 15 years old was 7.7%. The over-weight rate and the obesity rate for adults were 35.1% and 20.6%, respectively.

CONCLUSIONThe following risk factors of chronic disease, such as unreasonable diet pattern, deficient intake of some micronutrients, over-weight and obesity, lack of physical examination, smoking and over-drinking, should be the important factors influencing the health of Beijing residents severely.

China ; epidemiology ; Diabetes Mellitus ; epidemiology ; Female ; Health Status ; Humans ; Hypertension ; epidemiology ; Incidence ; Life Style ; Male ; Nutritional Status ; Obesity ; epidemiology ; Surveys and Questionnaires

OBJECTIVETo establish a mice model of aminoglycoside antibiotics (AmAn) induced ototoxicity. Then to investigate the sensitivity of AmAn induced ototoxicity in three mouse strains and effect of kanamycin on the expression of Na-K-2Cl co-transporter-1 (NKCC1) in stria vascularis.

METHODSC57BL/ 6J, CBA/CaJ, NKCC1 +/- mice (each of twenty-four) were randomly divided into four experimental groups A, B, C and D (A kanamycin alone, B kanamycin plus 2, 3-dihydroxybenzoate, C 2, 3-dihydroxybenzoate alone, D control group). Mice were injected with kanamycin or/and 2, 3-dihydroxybenzoate for 14 days. Auditory function was measured by auditory brainstem response (ABR) and morphology of cochlea was observed by succinate dehydrogenase staining. Expression of NKCC1 was detected by immunohistochemistry.

RESULTSMice in group A developed significant ABR threshold shifts (P < 0.01), which were accompanied by out hair cells loss. Mice in group B significantly attenuated ABR threshold shifts with out hair cells loss (P <0.01). The immunostaining of NKCC1 in stria vascularis was attenuated significantly in group A compared with group D (P < 0.01) while the immunostaining in group B was enhanced than which in group A (P < 0.01). CBA/CaJ mice has the highest sensitivity to AmAn in three mouse strains.

CONCLUSIONSAn mouse model of AmAn induced ototoxicity could be established by administration of kanamycin. Kanamycin could inhibit the expression of NKCC1 in stria vascularis. 2, 3-dihydroxybenzoate could attenuate AmAn induced ototoxicity maybe by enhancing the expression of NKCC1. Mice that had the characteristic of presbycusis didn't show additional sensitivity of AmAn induced ototoxicity.

Animals ; Anti-Bacterial Agents ; toxicity ; Blood Vessels ; drug effects ; metabolism ; Cochlea ; blood supply ; drug effects ; metabolism ; Kanamycin ; toxicity ; Mice ; Mice, Inbred C57BL ; Mice, Inbred CBA ; Parabens ; toxicity ; Sodium-Potassium-Chloride Symporters ; metabolism ; Solute Carrier Family 12, Member 2

BACKGROUNDSeveral isoforms of p53 have been reported, which may have varying functions and expressions. This study aimed to analyze the expression patterns of p53 isoforms in renal cell carcinoma (RCC) at the mRNA and protein levels and their associations with clinical and pathologic factors to explore the mechanism of p53 isoforms' activity in RCC.

METHODSThe specimens of tumours (T) and clinically normal tissues (N) adjacent to them were collected from 41 patients with RCC. mRNA expression levels of p53 isoforms were detected using RT-PCR followed by nested PCR. Protein expression levels were detected using immunohistochemisty and Western blotting with the anti-p53 antibodies DO-1 and DO-12. The data were analyzed with clinicopathological features by chi(2) test or Fisher's exact test.

RESULTSp53 mRNA was expressed in all tumours and matched clinically normal tissue adjacent to the tumour. All six isoforms could be detected in tumour and normal tissues, with the exception of the Delta133p53beta isoform, which was not detected in the normal tissue. Of the six isoforms, p53beta mRNA was significantly overexpressed in tumour samples (P < 0.001), and correlated with tumour stage. Nested PCR results consistently indicated the presence of p53gamma (19T/22N), Delta133p53 (33T/26N), Delta133p53beta (2T/0N), and Delta133p53gamma (13T/9N). Immunohistochemical analysis showed that p53 was expressed only in tumour tissues and correlated with tumour stage and grade. The results of Western blotting analysis were consistent with these findings.

CONCLUSIONSAlthough all six isoforms are present in RCC, their function in tumour development or progression might be different. Our findings suggest that p53beta might play an important role in the formation of RCC and it might be used as a new predictor and therapeutic target for RCC.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Carcinoma, Renal Cell ; genetics ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; genetics ; physiology ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Protein Isoforms ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Young Adult

OBJECTIVETo summarize the reasons for bile duct injury (BDI) after laparoscopic cholecystectomy (LC), and to determine the effect of multiple treatment after BDI.

METHODSA retrospective cohort study was performed. The medical records of 110 patients diagnosed with BDI after LC from October 1993 to November 2007, in ten large hospitals in Guangdong of China, were reviewed.

RESULTSAmong 110 patients with BDI, 58 cases (52.7%) were local patients, whereas 52 cases (47.3%) were transferred from outside hospitals. Reasons for BDI following LC were: (1) Lack of experience of the LC operator (48.2%); (2) LC performed during acute cholecystitis (20.0%); (3) The structure of Calot triangle was unclear (15.5%); (4) Variable anatomical position (11.8%); (5) Intra-operation bleeding (4.5%). The commonest sites of injury were the choledochus and common hepatic duct (76.4%). Following BDI, endoscopic stenting or operative repair was performed in 106 patients. The overall success rate was 95.3% (101/106), with a mortality rate was 0.9% (1/106). Cholangitis occurred in 3.8% (4/106) cases. Choledocho-enterostomy operation was performed in almost 60.0% (63/106) cases, and the success rate was 93.7% (59/63). Endoscopic stenting or operative repair was performed immediately following BDI in 23.6% (25/106) patients, the success rate was 100%; and within 30 days in 63.2% (67/106) patients. Eighty-eight out of 106 patients who underwent repair were successful following the first operative procedure.

CONCLUSIONSFactors such as an un-experienced operator and unclear anatomical position were causes of BDI following LC. Early operative repair should be regarded as the treatment of choice, in patients diagnosed with BDI. Early refer to an experienced hepatobiliary operator ensures a high success rate.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bile Ducts ; injuries ; surgery ; Cholecystectomy, Laparoscopic ; adverse effects ; Female ; Humans ; Iatrogenic Disease ; Intraoperative Complications ; diagnosis ; etiology ; surgery ; Male ; Middle Aged ; Retrospective Studies

Objective To investigate the characteristics of NEN metastasis with SSTR PET/CT and to correlate the results with FDG-PET and pathology.Methods From November 2011 to August 2016,a total of 43 patients with NEN (18 males,25 females;age range:26-74 years) were recruited into this retrospective study;they underwent 68Ga-DOTA-TATE PET/CT (TATE-PET) imaging 40-60 min after 44.4-229.4 MBq 68Ga-DOTA-TATE administration.Metastases in 31 patients were confirmed by histopathology and in 12 patients by follow-up and other imaging modalities.Twenty-eight of 43 patients finished routine FDG-PET in a week after TATE-PET.PET/CT results were considered positive when the metastatic lesions were tracer-avid.ROI was drawn over each lesion for size and SUV measurement.x2 test and Pearson correlation analysis were used for statistical analysis.Results (1) Of 43 patients,TATE-PET detection rates of metastasis in the liver,lymph nodes,bones.and lungs were 85.7％(30/35),12/13,7/7,2/3,respectively,with their corresponding SUVmax of 18.1(11.3-23.3),10.8(5.4-15.6),7.7(4.2-9.9) and 1.8(1.3-2.3),respectively.Statistical correlation between size and SUVmax was found in metastatic bone lesions(r=0.233,P＜0.05).(2) In 31 patients with histopathologically proven metastasis,TATE-PET detected 23/24 (95.8％) G2,1/2 G1 and 5/5 G3 metastases.G1 metastases were only found in the liver.(3)Among 28 patients underwent both TATE-PET and FDG-PET,there was no significant difference between the detecting rate of metastasis:89.2％ (25/28) vs 71.4％ (20/28);x2 =2.389,P＞0.05.Compared with FDG-PET,TATE-PET was superior in demonstrating metastasis in the liver and bones (70.0％ (14/20) vs 65.0％ (13/20),3/3 vs 2/3),equal in detecting lung metastasis (both 2/3) but inferior in demonstrating metastasis in lymph nodes (9/10vs 10/10).Conclusions The capability of TATE-PET in revealing NEN metastasis varies depending on lesion localization and histologic grade.TATE-PET and FDG-PET are complementary to each other in detection of NEN metastasis,but without obvious relationship to histologic grade.

OBJECTIVES: To investigate the clinical treatment outcomes and the changes of the outcomes over time in extremely preterm twins in Guangdong Province, China. METHODS: A retrospective analysis was performed for 269 pairs of extremely preterm twins with a gestational age of <28 weeks who were admitted to the department of neonatology in 26 grade A tertiary hospitals in Guangdong Province from January 2008 to December 2017. According to the admission time, they were divided into two groups: 2008-2012 and 2013-2017. Besides, each pair of twins was divided into the heavier infant and the lighter infant subgroups according to birth weight. The perinatal data of mothers and hospitalization data of neonates were collected. The survival rate of twins and the incidence rate of complications were compared between the 2008-2012 and 2013-2017 groups. RESULTS: Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of severe asphyxia and smaller head circumference at birth (P<0.05). The mortality rates of both of the twins, the heavier infant of the twins, and the lighter infant of the twins were lower in the 2013-2017 group compared with the 2008-2012 group (P<0.05). Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of pulmonary hemorrhage, patent ductus arteriosus (PDA), periventricular-intraventricular hemorrhage (P-IVH), and neonatal respiratory distress syndrome (NRDS) and a higher incidence rate of bronchopulmonary dysplasia (P<0.05). CONCLUSIONS There is a significant increase in the survival rate over time in extremely preterm twins with a gestational age of <28 weeks in the 26 grade A tertiary hospitals in Guangdong Province. The incidences of severe asphyxia, pulmonary hemorrhage, PDA, P-IVH, and NRDS decrease in both the heavier and lighter infants of the twins, but the incidence of bronchopulmonary dysplasia increases. With the improvement of diagnosis and treatment, the multidisciplinary collaboration between different fields of fetal medicine including prenatal diagnosis, obstetrics, and neonatology is needed in the future to jointly develop management strategies for twin pregnancy.
Bronchopulmonary Dysplasia/epidemiology* ; Female ; Gestational Age ; Humans ; Infant ; Infant, Extremely Premature ; Infant, Newborn ; Pregnancy ; Respiratory Distress Syndrome, Newborn/epidemiology* ; Retrospective Studies ; Treatment Outcome

Objective: The relationship between serum uric acid (SUA) levels and glycemic indices, including plasma glucose (FPG), 2-hour postload glucose (2h-PG), and glycated hemoglobin (HbA1c), remains inconclusive. We aimed to explore the associations between glycemic indices and SUA levels in the general Chinese population. Methods: The current study was a cross-sectional analysis using the first follow-up survey data from The China Cardiometabolic Disease and Cancer Cohort Study. A total of 105,922 community-dwelling adults aged ≥ 40 years underwent the oral glucose tolerance test and uric acid assessment. The nonlinear relationships between glycemic indices and SUA levels were explored using generalized additive models. Results: A total of 30,941 men and 62,361 women were eligible for the current analysis. Generalized additive models verified the inverted U-shaped association between glycemic indices and SUA levels, but with different inflection points in men and women. The thresholds for FPG, 2h-PG, and HbA1c for men and women were 6.5/8.0 mmol/L, 11.0/14.0 mmol/L, and 6.1/6.5, respectively (SUA levels increased with increasing glycemic indices before the inflection points and then eventually decreased with further increases in the glycemic indices). Conclusion An inverted U-shaped association was observed between major glycemic indices and uric acid levels in both sexes, while the inflection points were reached earlier in men than in women.
Aged ; Asian Continental Ancestry Group ; Blood Glucose/analysis* ; China/epidemiology* ; Cohort Studies ; Diabetes Mellitus/blood* ; Female ; Glucose Tolerance Test ; Glycated Hemoglobin A/analysis* ; Glycemic Index ; Humans ; Male ; Middle Aged ; Uric Acid/blood*