Objective To study the mutations at 1 573 fragment of TNF receptor II (TNFR-II) gene in patients with keloid. Methods The tissue DNA was extracted from 22 samples of keloids donated by 22 patients (6 males and 16 females, aged 18-53 years), and all keloids were examined and classified by pathologist. The peri pheral blood DNA was extracted from the same patients as the control. PCR was used to ampl ify the 1 573 fragment of TNFR-II gene from the keloid tissue DNA and peripheral blood DNA. The PCR products were sequenced directly and then compared with the GeneBankdata. Results All the concentration of the extracted DNA in trial were higher than 0.50 μg/μL and the purity (A260/A280) ofthe extracted DNA were higher than 1.5. It closed to the magnitude of the design DNA fragment by agarose gel electrophoresis examining, and corresponded with the test requirement. Mutations at 1 573 fragment of TNFR-II gene were detected in 13 out of 22 keloids. The mutation incidence was 59.1%. Among them, 9 had point mutation at codon 1 663, accounting 40.9%. No TNFR-II gene mutation was detected in all peripheral blood samples. There were significant difference between keloids DNA and peripheral blood DNA (P lt;0.01). The mutations involved point mutation, deletion and insertion as well as multisite and multitype. Conclusion There is a correlation between the mutation at 1 573 fragment of TNFR-II gene and keloid.
Microorganism distributes in the organs of human body which connect with external environment, especially those organs in the gastrointestinal tracts, and it also plays a fundamental role in the physiopathology of the host's body. Because the microorganism is very small and has a great variety, it is difficult to reveal the significance of microorganism in the human physiopathology comprehensively and deeply. With the development of molecular biology, genomics, bioinformatics and other disciplines, the microbiome research will be more possible and easier. There are two key contents of microecology. One of these is to identify and quantify the diversity of microorganism, and the other is to reveal activity and the physiopathological function of microorganism in the host. Microbiome research methods, therefore, can be summarized as the traditional detection methods, construction of gene library, the genetic fingerprint analysis and molecular hybridization techniques and so on.
Objective To investigate the histological origin, diagnosis, differential diagnosis and treatment of thyroid carcinoma showing thymus-like differentiation (CASTLE). Methods Five patients with thyroid CASTLE were adopted by surgical resection and postoperative radiotherapy, and the CD5, CD117, CK5/6, P63, thyroid transcription factor-1 (TTF-1), carcino-embryonic antigen (CEA), calcitonin (CT), Ki-67, chromogranin A (CgA), thyrobolulin (Tg), peroxisome proliferator activated receptorγ (PPAR-γ), sodium iodide symporter (NIS), and thyroid stimulating hormone receptor (TSHR) were detected in tumor tissues by immunohistochemistry S-P method and v-raf murine sarcoma viral oncogene homolog B1 (BRAF)V600E gene and telomerase reverse transcriptase (TERT) promoter mutations were detected by DNA sequencing. Eight cases of poorly differentiated thyroid carcinoma and 6 cases of anaplastic thyroid carcinoma were adopted by comprehensive comparative analysis. Results Thyroid CASTLE tumor cells showed the positive expression of CD5, CD117, CK5/6 and P63, and the negative expression of TTF-1, CT, CgA, Tg, PPAR-γ, NIS and TSHR. There were partly positive expression for CK5/6, P63, TTF-1, CgA, Tg, NIS and TSHR, and negative expression for CD5 and CD117 in the poorly differentiated thyroid carcinoma and anaplastic thyroid carcinoma. The BRAFV600E gene and TERT promoter mutations were not detected in thyroid CASTLE, and the BRAFV600E gene mutations were also not detected in the poorly differentiated thyroid carcinoma and anaplastic thyroid carcinoma. Four cases of poorly differentiated thyroid carcinoma showed the TERT promoter mutations (4/8) included 3 cases with C228T and 1 case with C250T. Two cases of anaplastic thyroid carcinoma showed the TERT promoter mutations (2/6) included 1 case with C228T and 1 case with C250T. There was no recurrence and metastasis after 3–47 months (an average of 25.6 months) of followed-up in thyroid CASTLE patients. Conclusions The histological origin of thyroid CASTLE may be not related to the thyroid. There is important clinical value to combined detection of CD5, CD117, P63, TTF-1, Tg, NIS, and TSHR for the diagnosis and differential diagnosis of thyroid CASTLE. The further study still need for the diagnosis and differential diagnosis of thyroid CASTLE according to the detection of BRAFV600E and TERT promoter mutations.
Objective To investigate the effect of cholecystolithiasis with cholecystitis and cholecystectomy on intestinal flora in patients with colorectal cancer. Methods A total of 168 patients with colorectal cancer who admitted to the Department of Anorectal Surgery in Gansu Provincial Hospital from June 2020 to March 2021 were selected, and 29 patients with colorectal cancer who met the criteria were selected as the research objects, including 10 colorectal cancer patients with gallstones and cholecystitis (cholecystolithiasis with cholecystitis+colorectal cancer group), 10 colorectal cancer patients after cholecystectomy (cholecystectomy+colorectal cancer group), and 9 colorectal cancer patients with normal gallbladder (normal gallbladder+colorectal cancer group). Clinical data of the patients in three groups were collected and compared. The fresh fecal samples of the patients included in the study were collected, and the 16S rDNA high-throughput sequencing method was used to determine and analyze the composition and distribution of the intestinal flora in the obtained samples. Results The interleukin-6 level in the cholecystolithiasis with cholecystitis+colorectal cancer group was statistically higher than that in the normal gallbladder+colorectal cancer group and the cholecystectomy+colorectal cancer group (P<0.05). At the phylum level of the fecal flora in three groups patients: ① In the samples of three groups, the relative abundances of Bacteroidetes, Firmicutes, Proteobacteria, Fusobacteria and Verrucomicrobia phylums were all high, accounting for almost more than 95% of the total intestinal bacteria. ② The relative abundance of Fusobacteria phylum in the cholecystolithiasis with cholecystitis+colorectal cancer group was statistically higher than that in the normal gallbladder+colorectal cancer group (P<0.05). ③ The relative abundance of Verrucomicrobia phylum in the normal gallbladder+colorectal cancer group was statistically higher than that in the cholecystolithiasis with cholecystitis+colorectal cancer group and the cholecystectomy+colorectal cancer group (P<0.05). ④ The relative abundance of Synergistetes phylum in the cholecystectomy+colorectal cancer group was statistically higher than that in the cholecystolithiasis with cholecystitis+colorectal cancer group and the normal gallbladder+colorectal cancer group (P<0.05). At the genus level: ① The relative abundances of Bacteroidetes and Roseburia genus were lower in the gallstone with cholecystitis+colorectal cancer group than those in the cholecystectomy+colorectal cancer group and the normal gallbladder+colorectal cancer group (P<0.05). ② The relative abundance of Shigella genus in the cholecystectomy+colorectal cancer group was higher than that in the cholecystolithiasis with cholecystitis+colorectal cancer group (P<0.05). ③ The relative abundance of the Lachnospira genus in the cholecystolithiasis with cholecystitis+colorectal cancer group was lower than that in the normal gallbladder+colorectal cancer group (P<0.05). ④ The relative abundances of Prevotella and Fusobacteria genus were higher in the cholecystolithiasis with cholecystitis+colorectal cancer group than that in the cholecystectomy+colorectal cancer group and the normal gallbladder+colorectal cancer group (P<0.05). ⑤ The relative abundances of Clostridium and Akkermansia genus were lower in the cholecystolithiasis with cholecystitis+colorectal cancer group and the cholecystectomy+colorectal cancer group than that in the normal gallbladder+colorectal cancer group (P<0.05). ⑥ The relative abundance of Enterococcus genus was higher in the normal gallbladder+colorectal cancer group than that in the cholecystectomy+colorectal cancer group (P<0.05).Conclusions ① Long-term occurrence of cholecystolithiasis with cholecystitis can cause obvious decrease in the abundances of Bacteroides, Roseburia, Lachnospira, etc. ② Cholecystectomy can cause changes in the relative abundances of Clostridium, Enterococcus, Verrucomicrobia, Synergistetes, etc. ③ The relative abundance of Fusobacterium is obviously increased in colorectal cancer patients with gallstones and cholecystitis, then promotes the release of inflammatory cytokines and causes intestinal inflammation, which is conducive to the growth of opportunistic pathogens, thus may affect the occurrence and development of colorectal cancer.
ObjectiveTo explore the composition of intestinal microbiota between patients with fixed airflow obstruction asthma, reversible airflow obstruction asthma, and healthy control, and analyze the correlation between key differential bacterial distribution and clinical characteristics. MethodsFifteen patients with fixed airflow obstruction asthma (FAO) and 13 patients with reversible airflow obstruction asthma (RAO) were included, along with 11 matched healthy control subjects. Clinical data were collected, and lung function tests and induced sputum examination were performed. Blood and stool samples were tested to compare the gut microbiota status among the groups, and analyze the relationship between gut microbiota abundance and patients' blood routine, IgE levels, lung function, and induced sputum. Results The dominant bacterial compositions were similar in the three groups, but there were differences in the abundance of some species. Compared to the RAO group, the FAO group showed a significant increase in the genera of Bacteroides and Escherichia coli, while Pseudomonas was significantly decreased. The phylum Firmicutes was negatively correlated with the course of asthma, while the phylum Bacteroidetes and genus Bacteroides were positively correlated with the asthma course. Bacteroidetes was negatively correlated with Pre-BD FEV1/FVC, Pseudomonas was positively correlated with Pre-BD FEV1, Escherichia coli was negatively correlated with Post-BD FEV1/FVC, and Bacteroides was negatively correlated with Post-BD MMEF. The class Actinobacteria and the order Actinomycetales were negatively correlated with peripheral blood EOS%, while the order Enterobacteriales and the family Enterobacteriaceae were positively correlated with peripheral blood IgE levels. Furthermore, Actinobacteria and Actinomycetales were negatively correlated with induced sputum EOS%. Conclusions There are differences in the gut microbiota among patients with fixed airflow obstruction asthma, reversible airflow obstruction asthma, and healthy individuals. Bacteroides and Escherichia coli are enriched in the fixed airflow obstruction asthma group, while the Firmicutes are increased in the reversible airflow obstruction asthma group. These three microbiota may act together on Th2 cell-mediated inflammatory responses, influencing the process of airway remodeling, and thereby interfering with the occurrence of fixed airflow obstruction in asthma.