Objective To explore the mechanisms of trafficking and signaling of serotonin 1A receptor(5-HT_(1A))and its spatiotemporal distribution in living cells.Methods The mouse 5-HT_(1A) gene amplified by RT-PCR was recombined into pEGFP-N1 vector and the EGFP coding sequence was located in-frame at the C-terminal end of the 5-HT_(1A) receptor.The 5-HT_(1A)-EGFP was transfected into neuron-like PC12 cells as well as HEK293.The transfected cells were visualized using confocal microscopy,the mobility of 5-HT_(1A)-EGFP was monitored by live measurements and fluorescence recovery after photobleaching.Results The 5-HT_(1A) gene was identitical with the published gene sequence NM_008308.4 and a 5-HT_(1A)-EGFP fusion construct was created.After stable transfection of the plasimd into a PC12 cell line and analysis with a confocal laser scanning microscopy,the EGFP-tagged 5-HT_(1A) was predominantly associated with the plasma membrane,but some intracellular vesicles in the perinuclear region also contained the fusion protein.The predominant localization of 5-HT_(1A)-EGFP at the plasma membrane was confirmed in transiently transfected HEK293 cells.Bleached fluorescence was partialy recovered in 100 seconds,indicating that the 5-HT_(1A)-EGFP was mobiled on the membrane.Conclusion Spatiotemporal distribution and mobility of 5-HT_(1A) tagged with EGFP can be monitored in the 5-HT_(1A)-EGFP stable PC12 cell line,which could be an excellent neuron-like experimental cell model for research of 5-HT_(1A) trafficking and signaling.

OBJECTIVE: To investigate the expression of signal transducer and activator of transcription 3 (STAT3) and phosphorylated STAT3 (p-STAT3) protein in papillary thyroid carcinoma (PTC), and to explore the correlation and significance between the expression of STAT3, p-STAT3 and epithelial-mesenchymal transition (EMT). METHOD: The expression of STATS3. p-STAT3, E-cadherin and Vimentin protein in 56 cases of PTC specimens and adjacent normal tissues specimens ware detected by immunohistochemistry. The correlation of the expression of STATS, p-STAT3, E-cadherin and Vimentin protein in PTC with clinicopathological characteristics was analyzed. RESULT: The positive rates of STAT3, p-STAT3 in PTC tissue were significantly higher than those in adjacent normal tissues specimens respectively (P < 0.01). The positive rates of E-cadherin in PTC tissues were remarkably lower, compared to adjacent normal tissues specimens (P < 0.01), however the positive rates of Vimentin in PTC tissues were remarkably higher, compared to adjacent normal tissues specimens (P < 0.01). The expression of STAT3, p-STAT3, E-cadherin and Vimentin protein were associated with lymph node metastasis and clinical stage (all P < 0.05). The expression of STAT3 and p-STAT3 was negatively correlated with E-cadherin expression (r = -0.494 and r = -0.364, P < 0.01, respectively), but positively with Vimentin expression (r = 0.533 and r = 0.377, P < 0.01, respectively) in PTC tissues. CONCLUSION PTC tissues have STAT3 protein activation and EMT phenotype, as were all correlated significantly with PTC invasion and metastasis. STAT3 signaling pathway activation might mediate EMT and then promote PTC invasion and metastasis.
Adult ; Antigens, CD ; Cadherins ; metabolism ; Carcinoma ; metabolism ; pathology ; Carcinoma, Papillary ; Epithelial-Mesenchymal Transition ; Female ; Humans ; Male ; Middle Aged ; Phosphorylation ; STAT3 Transcription Factor ; metabolism ; Thyroid Cancer, Papillary ; Thyroid Neoplasms ; metabolism ; pathology ; Vimentin ; metabolism

Objective To study the etiology and drug resistance of escherichia coli isolated from clinic in chengde from 2016 to 2018, and to provide an objective basis for the epidemiological study and clinical rational use of antimicrobial agents. Methods The feces of 702 diarrhea patients in an outpatient clinic in chengde district from 2016 to 2018 were collected, isolated and prepared by conventional pathogen testing procedures, and the specimens suspected of escherichia coli were identified by multiple PCR and single PCR experiments. DEC was sensitized to 15 antibiotic drugs in vitro by automatic microbiological identification and drug sensitivity analyzer Vitke-2 compact. Results A total of 189 DEC strains was isolated from the feces of 702 diarrhea patients, and the detection rate was 26.92%. Among 189 DEC strains, EAEC had the highest detection rate (57.67%), followed by ETEC (32.80%), then EPEC (6.35%) and mixed (3.17%). EIEC and EHEC were not detected. The virulence genes were mainly ast A 、est Ib and esc V. All types of DEC were detected throughout the year, with the highest detection rate (29.19%) between June and August. The patients were mainly aged between 20 and 45 years old, and the detection rate was 25.86% and 27.97% for men and women. The detection of EPEC was not affected by season, sex or age, except by season and age. DEC had the highest resistance to ampicillin at 62.96%, 51.32%, 49.74% and 30.16% to cotrimoxazole, ciprofloxacin and gentamicin, and less than 20% to the other 11 drugs, all of which were sensitive to meropenem but not resistant. Among 189 DEC strains, esbl-producing strains accounted for 1.59%. Multiple resistant strains accounted for 32.80%. Conclusion DEC for chengde area of the most common bacterial pathogens in gastrointestinal tract infection, clinical laboratory should strengthen the monitoring of the region DEC, especially to carry on a variety of virulence genes, ESBL producing and multiple drug resistance strains, timely grasp the dynamics of bacterial drug resistance, and to guide clinical rational drug use, delay and reduce the generation of bacterial drug resistance.

ObjectiveTo explore the correlation between blood stasis constitution （BSC） and diseases based on constitution literature involving “constitution-diseases correlation”. MethodsA comprehensive search was conducted on six Chinese and English electronic databases including CNKI， Wanfang， VIP， SinoMed， PubMed and Embase to find all clinical researches on the correlation between constitution and diseases using the Classification and Identification of Chinese Medicine Constitution standard from April 1st， 2009 to December 31st， 2022， and the participants of the research were BSC related. By analyzing the characteristics of the literature， such as authors， publication institutes， participants， and results， the disease with the highest proportion of BSC distribution or BSC as their risk factors or protective factors were summarized to explore the correlation between BSC and diseases. ResultsTotally 135 clinical studies on diseases highly related to BSC were included， with a total sample size of 71 172 cases.There were 27 keywords in the articles appeared more than 3 times， including the elderly， lumbar disc herniation， coronary heart disease， cardiovascular disease， and endometriosis. In the author's clustering， included studis were mainly from Shenzhen Hospital of Beijing University of Chinese Medicine， Second Affiliated Hospital of Guangzhou University of Chinese Medicine， and Wenzhou Central Hospital. In terms of blood stasis related diseases， 81 studies showed that BSC was the most common type of constitution in the study population， involving 48 disease or morbid states. The diseases and median proportions of BSC with reported literature ≥3 included coronary heart disease （28.8%）， endometriosis （31.3%）， neurocognitive impairment （26.4%）， lumbar disc herniation （26.0%）， ischemic stroke （25.0%）， adenomyosis （34.7%）， and endometrial polyps （25.0%）. Fifty-eight studies found that BSC was a risk factor for disease occurrence，and these diseases reported more than 3 times included hypertension （median OR = 2.956）， type 2 diabetes （median OR = 3.436），osteoporosis （median OR = 5.171）， sudden deafness （median OR = 3.827） and endometriosis （median OR = 5.412）. One study indicated BSC as the protective factor of lateral growth tumor of large intestine （median OR = 0.161）. ConclusionBSC is closely related to circulatory system diseases， urogenital system diseases， and musculoskeletal system diseases.