Objective: To investigate the change in intestinal flora in Mongolian students with anxiety,so as to provide basis for exploring the relationship between flora and secretion expression in vivo. Methods: The Self rating Anxiety Scale(SAS)was used to assess anxiety in medical college students; then a semi structured interview was administered. Fecal samples that met the inclusion criteria were collected and divided into anxiety (SAS score≥50) and control groups (no anxiety, SAS score<50) according to the standard score of SAS. Samples provided by Mongolian female students were selected from each group. The total bacterial DNA was extracted from the fecal samples for PCR amplification and NovaSeq 2x250bp high throughput sequencing was performed for the V3- V4 region of 16S rDNA gene to obtain the biological information of the intestinal flora. The intergroup OTU, structural diversity, significant difference, and LEfSe analyses were performed with information mining of the literature think tanks. Results: Anxiety existed in 23.86% of the Mongolian students,and 16.96% of the Han people. A Chi square test showed no significant difference in detection of anxiety between Mongolian and Han college students ( P =0.07). Analysis of the alpha diversity index showed that the Shannon index, Simpson index, Chao1 index, and Observed species did not differed significantly between the two groups( t =8.0， 9.0 ，6.0，6.5). The difference in abundance of some bacteria was significant at the Class, Order, Family, and Genus levels between the two groups( t =-2.26-2.57，-5.08-3.58，-2.65-2.09， P <0.05). Conclusion The alpha diversity index showed that there was no significant difference in the abundance and diversity of intestinal flora between the two groups. While there were significant differences at different classification levels, the results suggest that the structure of intestinal flora can change in students with anxiety.

Purpose To investigate the expression of TRPS1 in synovial sarcoma(SS)and its significance.Methods Twenty-one cases of synovial sarcoma diagnosed by SS1 8(18q11)(SYT)fluorescence in situ hybridization(FISH)or SS18-SSX immunohistochemistry were collected.The expression of TRPS1 was detected by immunohistochemistry EnVision two-step staining,and the relevant literature was reviewed.Results There were 21 cases of synovial sarcoma,including 9 females and 12 males,aged 17-79 years,with a median age of 50 years and an average age of 47.1 years.The tumors occurred in the lung(3 cases),legs(3 cases),kidney(3 cases),feet(2 cases),psoas major muscle(2 cases),rectum(1 case),tho-racic cavity(1 case),ankle canal(1 case),inguinal area(1 case),knee(1 case),palm(1 case),nasal cavity(1 case),and forearm(1 case).The maximum diameter of the tumor was 1.2-13 cm.Grossly,the tumor section was grayish-white,grayish-yellowish solid,and the lesion with hemorrhage was dark red.Microscopically,there were monophasic fibrous SS(16/21,76.2％),poorly differential SS(small cell)(3/21,14.3％),and biphasic SS(2/21,9.5％).Immunophenotypi-cally,tumor cells expressed SS18-SSX(18/18,100％),TLE1(12/12,100％),BCL2(18/18,100％),partially expressed EMA(10/16,62.5％),CK(12/18,66.7％),CD99(4/10,40.0％),SMA(3/16,18.6％),S-100(3/19,15.8％),and desmin(0/21).The proliferation index of Ki67 ranged from 3％-80％,with an average of 39.9％.FISH test showed that all 11 cases were positive for SS18(18q11)split-ting.All 21 cases of SS expressed TRPS1 to varying degrees and intensities(21/21,100％),of which 19 cases(19/21,90.5％)had a positive range of＞50％.There were 19 cases(19/21,90.5％)with a positive intensity of 2+or above.All the 21 patients underwent surgical resection of the tumor,and 20 patients received follow-up visits.3 patients treated with postop-erative combination of radiotherapy and chemotherapy,18 pa-tients treated with postoperative chemotherapy,and 7 patients showed different degrees of recurrence or metastasis,and 9 pa-tients who received follow-up visits died(45％,9/20).Con-clusion Although TRPS1 is regarded as a highly sensitive and specific marker of breast tumor origin,it is also highly expressed in synovial sarcoma,and it is necessary to be vigilant at its pit-falls in the pathological diagnosis.

We constructed a phage-displayed random mutation library of Tat38-61(51N/55N), for studying the molecular evolution screening of HIV-1 Tat38-61 epitope. We used primers containing the random nucleotide sequences, and introduced the random mutations at the sites of 51 and 55 amino acids coding sequences into full-length Tat sequences by overlapping PCR. With the randomly mutated full-length Tat as template, the Tat38-61(51N/55N) mutants which contained recognition sequences for the Xba I in both ends were amplified by PCR using the designed primers. The mutants were cloned into Xba I site in the phagemid vector pCANTAB5S, then the recombinants were transformed into E. coli TG1, a phage-displayed the random mutation library of Tat38-61(51N/55N) was constructed by the rescue of help virus M13KO7. The results showed that the library consisted of about 5.0 x 10(6) colonies and the phage library titer was 2.65 x 10(12) TU/mL. More than 56.50% colonies in the library were positive for insertion. Sequence analysis showed that the nucleotides encoding amino acids at the sites of 51 and 55 distributed randomly. The constructed mutation library could meet the requirements for the following molecular evolution screening, and might prepare the Tat mutants for the further study of new Tat vaccine candidates.
AIDS Vaccines ; immunology ; Escherichia coli ; genetics ; metabolism ; HIV-1 ; genetics ; Humans ; Mutation ; Peptide Fragments ; biosynthesis ; genetics ; immunology ; Peptide Library ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; tat Gene Products, Human Immunodeficiency Virus ; biosynthesis ; genetics ; immunology

Objective:To study the clinical features, diagnosis and treatment of vagal paraganglioma in parapharyngeal space.Methods:Nine cases with vagal paraganglioma in parapharyngeal space were retrospectively analyzed who were diagnosed and treated between January 2006 and December 2018 in Department of Otorhinolaryngology Head and Neck Surgery, Beijing Friendship Hospital and the First Medical Center, Chinese PLA General Hospital. There were 6 males and 3 females, aged from 24 to 50 years old. The main symptoms in the 9 patients were hoarseness and neck mass, and the secondary symptoms were irritating cough, cough on drinking and dysphagia. The main sign was a well-circumscribed round mass, tough in texture, with or without ipsilateral lateral oropharyngeal wall uplift and vocal cord paralysis. The tumors were located between the bifurcation of the carotid artery and the jugular foramen in 7 cases and intruded into jugular fossa in 2 cases. All the 9 patients underwent head and neck enhancement CT and MRI and 7 cases received digital subtraction angiography (DSA) examination and balloon occlusion test. The imaging features were tumors with rich blood supply in the parapharyngeal space of the upper neck, and the tumors were heterogeneous enhanced with contrast CT scan and enhanced MRI, which were closely related to the internal carotid artery, external carotid artery and jugular vein.Results:Among these 9 patients, 8 underwent surgical resection of tumors, including complete tumor resection in 7 cases and partial tumor resection in 2 case. One patient underwent partial tumor resection after being transferred to vascular surgery. There was no recurrence in 7 patients with complete tumor resection and slow growth was shown in 2 patients with partial tumor resection. Posterior cranial nerve injury occurred in 2 patients and stroke in 1 patient due to intraoperative ligation of internal carotid artery.Conclusions:Vagal paraganglioma in the parapharyngeal space is rich in blood supply and closely related to the internal and external carotid arteries, internal jugular vein and posterior cranial nerves. Surgical resection is the first choice for treatments. Choosing a reasonable operative approach for fully exposing the operative field and completely removing the tumor while protecting the internal carotid artery are the keys to successful surgery.

Objective:To explore the effect and correlation of long non-coding RNA (lncRNA) IDI2-AS1/microRNA-33b-5p (miR-33b-5p)/nuclear receptor-associated protein NR4A2 competitive endogenous RNA (ceRNA) regulatory network on acute myocardial infarction (AMI), and to verify whether IDI2-AS1 regulates NR4A2 through miR-33b-5p to affect the occurrence and development of myocardial infarction.Methods:The miRNA and mRNA expression chips related to myocardial infarction were obtained from gene expression omnibus (GEO), and the differential expression was analyzed. The upstream regulatory mechanism of NR4A2 was predicted using TargetScan database. Thirty-two male C57/BL6 mice were divided into Sham group, AMI model group, miR-33b-5p mimic group [miR-33b-5p mimic lentivirus (5×10 7 TU) was injected locally into the heart tissue during ligation] and miR-33b-5p inhibitor group [miR-33b-5p inhibitor lentivirus (5×10 7 TU) was injected locally into the heart tissue during ligation] according to random number table method, with 8 mice per group. Left ventricular end-diastolic diameter (LVEDD) and left ventricular end-systolic diameter (LVESD) were asseessed by echocardiography, left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) were calculated. After the last weighing, the anesthetized mice were sacrificed and the heart tissues were taken. Masson staining of the heart tissues was observed under light microscope, myocardial collagen volume fraction (CVF) and infarct size were calculated. Cardiomyocytes of SPF grade SD rats were collected. They were divided into normal control group (control group), ischemia-hypoxia model group, miR-33b-5p mimic transfection group (miR-33b-5p mimic transfection group before ischemia and hypoxia treatment) and miR-33b-5p inhibitor transfection group (miR-33b-5p inhibitor transfection group before ischemia and hypoxia treatment). The activity of caspase-3/7 in cardiomyocytes was measured. The levels of interleukins (IL-1β, IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). The levels of malondialdehyde (MDA), superoxide dismutase (SOD), creatine kinase (CK), MB isoenzyme of creatine kinase (CK-MB) and lactate dehydrogenase (LDH) were detected by colorimetry. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of apoptosis-related proteins Bax and Bcl-2, cytochrome C (Cyt C) and IDI2-AS1/miR-33b-5p/NR4A2 regulatory axis genes. Results:The myocardial infarction microarray analysis showed that NR4A2 expression was significantly up-regulated in myocardial infarction, with predicted upstream regulatory mechanisms indicating its possible influence through the IDI2-AS1/miR-33b-5p/NR4A2 regulatory axis. Echocardiographic detection showed that compared with AMI model group and miR-33b-5p inhibitor group, LVEF and LVFS in the heart tissue of mice in miR-33b-5p mimic group were significantly increased, while the levels of LVEDD, LVESD, CK, CK-MB and LDH were significantly decreased, with statistical significance. Light microscope showed myocardial fibrosis and myocardial infarction in AMI model group and miR-33b-5p inhibitor group. In the miR-33b-5p mimic group, the degree of myocardial fibrosis was decreased and the myocardial infarction size was significantly reduced. Compared with AMI model group and miR-33b-5p inhibitor group, the levels of MDA, IL-1β, IL-6, TNF-α and the expressions of Bax and Cyt C in the heart tissue of mice in miR-33b-5p mimic group were significantly decreased, while the levels of SOD and Bcl-2 expression were significantly increased, and the differences were statistically significant. The expressions of IDI2-AS1 and NR4A2 in the heart tissue of mice in miR-33b-5p mimic group were significantly lower than those in AMI model group and miR-33b-5p inhibitor group [IDI2-AS1 (2 -ΔΔCt): 1.96±0.08 vs. 2.73±0.08, 3.10±0.05, NR4A2 (2 -ΔΔCt): 2.36±0.07 vs. 3.16±0.08, 3.80±0.08, all P < 0.01]. The expression of miR-33b-5p was significantly higher than that of AMI model group and miR-33b-5p inhibitor group (2 -ΔΔCt: 0.88±0.07 vs. 0.57±0.07, 0.23±0.01, both P < 0.01). The cell experiment results showed that the caspase-3/7 activity of rat neonatal cardiomyocytes in the miR-33b-5p mimic transfection group was significantly lower than that in the ischemia-hypoxia model group and the miR-33b-5p inhibitor transfection group, suggesting that miR-33b-5p can significantly reduce the apoptosis level of the ischemia-hypoxia model. The levels of peroxidation and inflammation indexes, important genes of apoptosis pathway and the expression of IDI2-AS1/miR-33b-5p/NR4A2 regulatory axis of rat neonatal cardiomyocytes in all groups were consistent with the above. Conclusion:IDI2-AS1 can regulate NR4A2 through miR-33b-5p, thus affecting the occurrence and development of AMI.

This study aims to develop an improved cell screening system for farnesoid X receptor (FXR) agonists based on a dual luciferase reporter gene system. FXR response element (FXRE) fragments from FXR target genes were cloned and inserted into upstream of firefly luciferase (Luc) gene in the plasmid pGL4-luc2P-Hygro. In combination with the internal reference plasmid containing renilla luciferase, a dual luciferase reporter gene system was developed and used for high throughput screening of FXR agonists. After studying the effects of over-expression of RXR, mouse or human FXR, various FXRE fragments, and different ratio of FXR plasmid amount to reporter gene plasmid, induction efficiency of the screening system was optimized by the known FXR agonist GW4064, and Z factor for the system reached 0.83 under optimized conditions. In summary, an improved cell screening system based on double luciferase reporter gene detection system was developed to facilitate the discovery of FXR agonists, where a new enhanced FXRE element was formed by a superposition of multiple FXRE fragments from FXR target genes, instead of a superposition of traditional IR-1 (inverted repeats-1) fragments.
Humans ; Mice ; Animals ; Transcription Factors/genetics* ; DNA-Binding Proteins/genetics* ; Receptors, Cytoplasmic and Nuclear/genetics* ; Genes, Reporter ; Luciferases/genetics*