[Upregulating KLF11 ameliorates intestinal inflammation in mice with 2, 4, 6-trinitrobenesulfonic acid-induced colitis by inhibiting the JAK2/STAT3 signaling pathway].

OBJECTIVE: To investigate the expression level of Kruppel-like transcription factor family member KLF11 in intestinal mucosal tissues of Crohn's disease (CD) and its regulatory effect on intestinal inflammation in CD-like colitis. METHODS: We examined KLF11 expression levels in diseased and normal colon mucosal tissues from 12 CD patients and 12 patients with colorectal cancer using immunofluorescence staining. KLF11 expression was also detected in the colon mucosal tissues of a mouse model of 2, 4, 6-trinitrobenesulfonic acid (TNBS)-induced colitis. A recombinant adenoviral vector was used to upregulate KLF11 expression in the mouse models and the changes in intestinal inflammation was observed. A Caco-2 cell model with stable KLF11 overexpression was constructed by lentiviral infection. The effect of KLF11 overexpression on expressions of JAK2/STAT3 signaling pathway proteins was investigated using immunoblotting in both the mouse and cell models. The mouse models were treated with coumermycin A1, a JAK2/STAT3 signaling pathway agonist, and the changes in intestinal inflammatory responses were observed. RESULTS: The expression level of KLF11 was significantly lowered in both the clinical specimens of diseased colon mucosal tissues and the colon tissues of mice with TNBS-induced colitis (P < 0.05). Adenovirus-mediated upregulation of KLF11 significantly improved intestinal inflammation and reduced the expression levels of inflammatory factors in the intestinal mucosa of the colitis mouse models (P < 0.05). Overexpression of KLF11 significantly inhibited the expression levels of p-JAK2 and p-STAT3 in intestinal mucosal tissues of the mouse models and in Caco-2 cells (P < 0.05). Treatment with coumermycin A1 obviously inhibited the effect of KLF11 upregulation for improving colitis and significantly increased the expression levels of inflammatory factors in the intestinal mucosa of the mouse models (P < 0.05). CONCLUSION: KLF11 is downregulated in the intestinal mucosa in CD, and upregulation of KLF11 can improve intestinal inflammation and reduce the production of inflammatory factors probably by inhibiting the JAK2/STAT3 signaling pathway.

[Clinical features of peripheral neuropathy with livedo reticularis: an analysis of seven cases].

The clinical characteristics, auxiliary examinations, skin and neuropathological features of 7 patients who had reticular cyanosis with peripheral neuropathy from the Department of Neurology, Huashan Hospital, Fudan University from January 2019 to December 2022 were retrospectively analyzed. Among the 7 patients, 5 were female and 2 were male.The age of onset of peripheral neuropathy was (39.8±21.3) years and the disease duration of peripheral neuropathy was (2.7±2.3) years. Three patients had acute onset and 4 patients had chronic onset. All the patients had limb numbness, with limb weakness in 6 patients and pain in 5 cases. Neuroelectrophysiological examination revealed 1 case of mononeuropathy, 2 cases of polyneuropathy, 2 cases of peripheral neuropathy, and 2 cases of sensory neuron neuropathy. Skin biopsy was performed in 3 patients, which presented hyperplasia and expansion of blood vessels in the dermis with lymphocyte infiltration. Nerve biopsy was performed in 3 patients, indicating axonal damage. Reticular cyanosis with peripheral neuropathy characterizes with numbness and weakness of limbs, most of which were accompanied by pain. Electrophysiological changes are in various forms. The pathological changes are dominated by the damage of axonal.

[High expression of MYH9 inhibits apoptosis of non-small cell lung cancer cells through activating the AKT/c-Myc pathway].

OBJECTIVE: To investigate the role of myosin heavy chain 9 (MYH9) in regulation of cell proliferation, apoptosis, and cisplatin sensitivity of non-small cell lung cancer (NSCLC). METHODS: Six NSCLC cell lines (A549, H1299, H1975, SPCA1, H322, and H460) and a normal bronchial epithelial cell line (16HBE) were examined for MYH9 expression using Western blotting. Immunohistochemical staining was used to detect MYH9 expression in a tissue microarray containing 49 NSCLC and 43 adjacent tissue specimens. MYH9 knockout cell models were established in H1299 and H1975 cells using CRISPR/Cas9 technology, and the changes in cell proliferation cell were assessed using cell counting kit-8 (CCK8) and clone formation assays; Western blotting and flow cytometry were used to detect apoptosis of the cell models, and cisplatin sensitivity of the cells was evaluated using IC50 assay. The growth of tumor xenografts derived from NSCLC with or without MYH9 knockout was observed in nude mice. RESULTS: MYH9 expression was significantly upregulated in NSCLC (P < 0.001), and the patients with high MYH9 expression had a significantly shorter survival time (P=0.023). In cultured NSCLC cells, MYH9 knockout obviously inhibited cell proliferation (P < 0.001), promoted cell apoptosis (P < 0.05), and increased their chemosensitivity of cisplatin. In the tumor-bearing mouse models, the NSCLC cells with MYH9 knockout showed a significantly lower growth rate (P < 0.05). Western blotting showed that MYH9 knockout inactivated the AKT/c- Myc axis (P < 0.05) to inhibit the expression of BCL2- like protein 1 (P < 0.05), promoted the expression of BH3- interacting domain death agonist and the apoptosis regulator BAX (P < 0.05), and activated apoptosis-related proteins caspase-3 and caspase-9 (P < 0.05). CONCLUSION: High expression of MYH9 contributes to NSCLC progression by inhibiting cell apoptosis via activating the AKT/c-Myc axis.

[Burden of non-communicable diseases attributable to population aging in China, 1990‒2050].

Objective: The direction and intensity of population aging on the burden of non-communicable diseases (NCDs) in China from 1990 to 2019 were analyzed, and the burden of NCDs in 2050 was predicted. Methods: The disease-specific disability-adjusted life years (DALYs), years of life lost (YLLs), and years lived with disability (YLDs) in the Chinese population from 1990 to 2019 were obtained from the Global Burden of Disease Study.The differences in indicators from 1990 to 2019 were attributed to the contribution of age structure, population size, and all other causes. The Bayesian age-time-cohort models were used to predict DALYs from NCDs to 2050. Results: The absolute level of DALYs caused by NCDs increased by 7.460 million from 1990 to 2019, and the age structure contributed 186.0% (95% Uncertainty Intervals (UIs): 178.4%-193.6%), population size contributed 77.0% (95% UIs: 69.5%-80.8%), all other causes contributed -163.0% (95% UIs:-163.1%- -159.3%). DALYs caused by NCDs consist of 2.527 million YLLs and 4.934 million YLDs, in which the contribution of age structure to YLLs and YLDs was 414.6% (95% UIs: 396.2%-432.5%) and 69.1% (95% UIs: 66.7%-71.4%), respectively. From 2019 to 2050, the diseases with increased DALYs due to changes in age structure are cardiovascular diseases, neoplasms, chronic respiratory diseases, neurological disorders, sense organ diseases, diabetes and kidney diseases, musculoskeletal disorders, digestive diseases, mental disorders, and skin and subcutaneous diseases in descending order. Conclusions: From 1990 to 2019, except for skin and subcutaneous diseases, the burden of other NCDs attributable to population aging increased, mainly due to disability. By 2050, the burden of NCDsattributable to population aging will continue to rise.

[Comparative analysis on prevalence of tobacco and e-cigarettes uses in junior middle school students in Shanghai, 2013 and 2019].

Objective: To examine the prevalence and trend of tobacco and e-cigarettes uses and identify the influencing factors for smoking behavior in junior middle school students in Shanghai, and provide data support and scientific basis for the development of tobacco control intervention strategy in adolescents. Methods: Multi-stage stratified random sampling method was used to select junior middle school students in 8 districts and 10 districts in Shanghai in 2013 and in 2019 respectively. Information about tobacco and e-cigarettes uses in the students were collected by using self-administrated questionnaire. The prevalence of tobacco and e-cigarettes uses were calculated, the difference between two years was compared with χ2 test. The influencing factors were identified by multivariate logistic regression analysis. Results: In 2019, the current smoking rate was 0.6% in junior middle school students in Shanghai, and the smoking attempt rate was 2.9%, both were lower than the levels in 2013 (0.7% and 6.9%). The current use rate of e-cigarettes was 0.6% in 2019,with no significant change compared with 2013 (0.6%). The proportion of the students who had heard of e-cigarettes in 2019 (78.4%) was higher than that in 2013 (47.2%). In 2019, the second-hand smoke (SHS) exposure rate at home, in both indoor and outdoor public places and on public transportations was 72.5%, which was slightly lower than the level in 2013 (73.0%), the differences were all significant (P<0.05). In 2019, the students seeing close friend smoking (OR=27.381, 95%CI: 12.037-62.287), seeing someone smoking in school (OR=2.477, 95%CI: 1.155-5.312), believing that SHS may not be harmful (OR=8.471, 95%CI: 1.464-49.005) had higher possibility of smoking. Being aged ≥15 years (compared with being aged ≤12 years, OR=8.688, 95%CI: 1.922-39.266), exposure to SHS in outdoor public place (OR=8.608, 95%CI: 1.048-70.692), close friend smoking (OR=8.115, 95%CI: 1.754-37.545) were positively associated with e-cigarettes use, and believing that smoking results in uncomfortable social contact [compared with believing that smoking results in comfortable social contact (OR=0.105,95%CI: 0.018-0.615)] were negatively associated with e-cigarettes use, the difference was significant (P<0.05). Conclusion: The prevalence of tobacco and e-cigarette uses in junior middle school students in Shanghai remained at a low level in recent years. The SHS exposure rate in junior middle school students is high. Smoking behavior of junior middle school students is closely related to personal attitude and awareness of tobacco, exposure to SHS, peer smoking and the situation of tobacco control in schools. Prevention and intervention should be carried out from multi-dimensions to effectively protect teenagers from tobacco hazards.

[The predictive value of ureteral wall area for impacted ureteral stones].

Objective: To investigate the clinical indicators for preoperative prediction of impacted ureteral stones and analyze the predictive value of ureteral wall area(UWA). Methods: A total of 197 patients who underwent ureteroscopic lithotripsy due to ureteral stones at our institution from January to December 2020 were retrospectively analyzed. Preoperative patient age, gender, body mass index (BMI), history of hypertension, diabetes mellitus, side of stone, location of stone, maximum diameter of stone, CT value of stone, C-reactive protein (CRP), creatinine, renal pelvis diameter, ureteral wall thickness and UWA were collected. Patients were divided into impacted and non-impacted groups according to whether the stones were impacted intraoperatively. Univariate analysis was used to compare the differences in each clinical indicator between the two groups, and multivariate logistic regression was performed to analyze the independent predictors of impacted stones for those with differences. The receiver operating characteristic (ROC) curve was used to analyze the predictive power of each independent predictor, and the Delong test was used to analyze whether the difference in the area under the curve (AUC) of each independent predictor was statistically significant. Results: All 197 patients successfully completed the operation, aged 51 (36, 56) years; 137 males and 60 females. According to the results of ureteroscopy, they were divided into 82 cases of impacted ureteral stones and 115 cases of non-impacted ureteral stones. Univariate analysis showed that there were significant differences in maximum stone diameter, stone CT value, renal pelvis diameter, ureteral wall thickness and ureteral wall area between the two groups (P<0.05); There was no significant difference in age, gender, BMI, history of hypertension, diabetes, stone side, location of stone, CRP and creatinine (P>0.05). Multivariate logistic regression analysis showed that stone CT value (P<0.01), ureteral wall thickness (P<0.001) and ureteral wall area were independent predictors of impacted ureteral stones (P<0.001). The ROC curve was used to compare the predictive efficacy of independent predictors of stone CT value, ureteral wall thickness and ureteral wall area. The area under the ureteral wall area curve was the largest (AUC = 0.901, 95%CI: 0.859-0.943, P<0.001), followed by ureteral wall thickness (AUC = 0.799, 95%CI: 0.736-0.862, P<0.001) and stone CT value (AUC = 0.700, 95%CI: 0.626-0.775, P<0.001). By Delong test, there were significant differences in AUC between ureteral wall area and stone CT value (Z=4.527, P<0.001) and ureteral wall thickness (Z=3.407, P<0.001). The best predictive value of ureteral wall area was 79.6 mm2. The sensitivity and specificity of this critical value for predicting ureteral incarcerated calculi were 80.1% and 89.5%. Conclusions: The UWA, ureteral wall thickness as well as the CT value of stones were all independent predictors of impacted ureteral stones, and UWA had a better predictive value.

Impact of age and menopausal stage on serum anti-Müllerian hormone levels in middle-aged women.

OBJECTIVE: This study aimed to evaluate the association between age, menopausal stage and serum anti-Müllerian hormone (AMH) levels in middle-aged women. METHODS: In this cross-sectional study, the serum AMH levels of 288 healthy women aged 40-55 years (divided into age groups: 40-44, 45-49 and 50-55 years) were evaluated. Stages of Reproductive Aging Workshop + 10 criteria were used to categorize these women into menopausal stages: late reproductive, menopausal transition and early postmenopausal stages. The impact of age, menopausal stage and hormone replacement therapy on serum AMH levels was analyzed using multi-factor analysis of variance. Effects of body mass index, smoking status and oral contraceptive use were simultaneously considered. RESULTS: The median AMH level was 0.140 ng/ml. Log-AMH levels varied according to age group (variance = 20.113, F = 88.538, p < 0.001) and menopausal stage (variance = 5.543, F = 24.501, p < 0.001). An exponential model defined as AMH = 227,421.757 × e(-0.301 × age) was fit to describe the decline in AMH level with age. The 5th-95th percentiles of the AMH levels ranged from less than 0.020 to 3.150, less than 0.020 to 1.944 and less than 0.020 to 0.030 ng/ml in the aforementioned menopausal stages, respectively. CONCLUSION: Age and menopausal stage were associated with AMH levels; age had a greater impact on AMH than menopausal stage in middle-aged women.

[Analysis of a pedigree with inherited factor V deficiency caused by compound heterozygous mutation].

Objective: To explore the molecular pathogenesis of a family with hereditary factor Ⅴ (FⅤ) deficiency. Methods: All the exons, flanking sequences, 5' and 3' untranslated regions of the F5 of the proband, and the corresponding mutation sites of the family members were analyzed via direct DNA sequencing. The CAT measurement was used to detect the amount of thrombin produced. The ClustalX software was used to analyze the conservation of mutation sites. The online bioinformatics software, Mutation Taster, PolyPhen-2, PROVEAN, LRT, and SIFT were applied to predict the effects of mutation sites on protein function. The Swiss-PdbViewer software was used to analyze the changes in the protein model and intermolecular force before and after amino acid variation. Results: The proband had a heterozygous missense mutation c.1258G>T (p.Gly392Cys) in exon 8 of the F5, and a heterozygous deletion mutation c.4797delG (p.Glu1572Lys fsX19) in exon 14, which results in a frameshift and produces a truncated protein. Her grandfather and father had p.Gly392Cys heterozygous variation, whereas her maternal grandmother, mother, little aunt, and cousin all had p.Glu1572LysfsX19 heterozygous variation. The ratio of proband's thrombin generation delay to peak time was significantly increased. Conservation analysis results showed that p.Gly392 was located in a conserved region among the 10 homologous species. Five online bioinformatics software predicted that p.Gly392Cys was pathogenic, and Mutation Taster also predicted p.Glu1572Lys fsX19 as a pathogenic variant. Protein model analysis showed that the replacement of Gly392 by Cys392 can lead to the extension of the original hydrogen bond and the formation of a new steric hindrance, which affected the stability of the protein structure. Conclusion: The c.1258G>T heterozygous missense mutation in exon 8 and the c.4797delG heterozygous deletion mutation in exon 14 of the F5 may be responsible for the decrease of FⅤ levels in this family.

MiR-204 reduces apoptosis in rats with myocardial infarction by targeting SIRT1/p53 signaling pathway.

OBJECTIVE: The aim of this study was to investigate the influence of micro ribonucleic acid (miR)-204 on rats with myocardial infarction by targeting the silent information regulator 1 (SIRT1)/p53 signaling pathway. MATERIALS AND METHODS: A total of 36 Sprague-Dawley rats were randomly divided into three groups, including: sham-operation group (n=12), model group (n=12) and miR-204 mimics group (n=12). The rats in the sham-operation group only underwent thoracotomy, without myocardial infarction injury. Meanwhile, the rats in model group and miR-204 mimics group were utilized to establish the models of myocardial infarction, and then, intervened with normal saline and miR-204 mimics, respectively. The morphology of myocardial tissues was observed via hematoxylin-eosin (HE) staining. Immunofluorescence was performed to detect the expression of Caspase-3. Target genes of miR-204 were analyzed using bioanalysis software. Western blotting (WB) assay was applied to measure the relative protein expression of SIRT1. MiR-204 expression and the messenger RNA (mRNA) expressions of SIRT1 and p53 were measured via quantitative Polymerase Chain Reaction (qPCR). Furthermore, cell apoptosis was determined through terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. RESULTS: HE staining showed that the morphology of myocardial tissues was normal in sham-operation group. Severe myocardial tissue injury was visible in model group, and the injury was relieved in miR-204 mimics group when compared with model group. The results manifested that the positive expression of Caspase-3 in cardiac tissues increased remarkably in the model group and miR-204 mimics group in comparison with sham-operation group (p<0.05). Meanwhile, it was evidently lower in miR-204 mimics group than model group (p<0.05). Based on the analysis via bioanalysis software, SIRT1 was the target gene of miR-204. WB results revealed that the relative protein expression level of SIRT1 was elevated notably in the other two groups compared with the 2sham-operation group (p<0.05). However, it was markedly lowered in miR-204 mimics group in contrast with model group (p<0.05). QRT-PCR results demonstrated that the model group and miR-204 mimics group exhibited distinctly lower expression of miR-204 but higher mRNA expressions of SIRT1 and p53 than sham-operation group (p<0.05). However, miR-204 mimics group exhibited prominently higher expression of miR-204 but lower mRNA expressions of SIRT1 and p53 than model group (p<0.05). Finally, the results of TUNEL assay demonstrated that the apoptosis rate increased remarkably in the model group and miR-204 mimics group when compared with sham-operation group (p<0.05). However, it decreased notably in miR-204 mimics group in comparison with model group (p<0.05). CONCLUSIONS: MiR-204 reduces the apoptosis level in rats with myocardial infarction via targeted inhibition of the SIRT1/p53 signaling pathway.

Orf Virus-Based Therapeutic Vaccine for Treatment of Papillomavirus-Induced Tumors.

Orf virus (ORFV) represents a suitable vector for the generation of efficient, prophylactic antiviral vaccines against different pathogens. The present study investigated for the first time the therapeutic application of ORFV vector-based vaccines against tumors induced by cottontail rabbit papillomavirus (CRPV). ORFV-CRPV recombinants were constructed expressing the early CRPV gene E1, E2, E7, or LE6. In two independent experiments we used in total 23 rabbits which were immunized with a mixture of the four ORFV-CRPV recombinants or empty ORFV vector as a control 5 weeks after the appearance of skin tumors. For the determination of the therapeutic efficacy, the subsequent growth of the tumors was recorded. In the first experiment, we could demonstrate that three immunizations of rabbits with high tumor burden with the combined four ORFV-CRPV recombinants resulted in significant growth retardation of the tumors compared to the control. A second experiment was performed to test the therapeutic effect of 5 doses of the combined vaccine in rabbits with a lower tumor burden than in nonimmunized rabbits. Tumor growth was significantly reduced after immunization, and one vaccinated rabbit even displayed complete tumor regression until the end of the observation period at 26 weeks. Results of delayed-type hypersensitivity (DTH) skin tests suggest the induction of a cellular immune response mediated by the ORFV-CRPV vaccine. The data presented show for the first time a therapeutic potential of the ORFV vector platform and encourage further studies for the development of a therapeutic vaccine against virus-induced tumors.IMPORTANCE Viral vectors are widely used for the development of therapeutic vaccines for the treatment of tumors. In our study we have used Orf virus (ORFV) strain D1701-V for the generation of recombinant vaccines expressing cottontail rabbit papillomavirus (CRPV) early proteins E1, E2, LE6, and E7. The therapeutic efficacy of the ORFV-CRPV vaccines was evaluated in two independent experiments using the outbred CRPV rabbit model. In both experiments the immunization achieved significant suppression of tumor growth. In total, 84.6% of all outbred animals benefited from the ORFV-CRPV vaccination, showing reduction in tumor size and significant tumor growth inhibition, including one animal with complete tumor regression without recurrence.

Sevoflurane improves circulatory function and pulmonary fibrosis in rats with pulmonary arterial hypertension through inhibiting NF-κB signaling pathway.

OBJECTIVE: The aim of this study was to explore the influences of sevoflurane inhalation therapy on circulation function and pulmonary fibrosis in rats with pulmonary arterial hypertension (PAH) and the nuclear factor-κB (NF-κB) signaling pathway. MATERIALS AND METHODS: A total of 30 adult male Sprague-Dawley rats were randomly divided into three groups, including control group (CTL group, n=10), PAH group (n=10), and PAH + sevoflurane group (n=10) using a random number table. Subsequently, the pulmonary artery right ventricular systolic pressure (RVSP) and right ventricular hypertrophy index (RVHI) were measured. Rats in PAH group were subcutaneously injected with 60 mg/kg monocrotaline once to establish the model of PAH. 28 d later, the differences in the morphology of pulmonary tissues and the protein expression levels of phosphorylated inhibitory κB (p-IκB), p-P65, P65, cyclin D1, proliferating cell nuclear antigen (PCNA) and tubulin among the three groups were analyzed via hematoxylin-eosin (HE) staining and Western blotting, respectively. Meanwhile, the messenger ribonucleic acid (mRNA) expression level of P65 was determined via Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR). Additionally, changes in the expression levels of Ki-67 and α-smooth muscle actin (α-SMA) in rat pulmonary tissues of the three groups were evaluated through immunohistochemistry. RESULTS: According to HE staining results, compared with CTL group, rats in PAH group exhibited significant thickening of the pulmonary artery wall, reduction of the vascular lumen, inflammatory cell infiltration, and thrombosis in some small arteries. This indicated that the PAH model was successfully established in rats. Compared with PAH group, PAH + sevoflurane group showed a significantly improved morphology of rat pulmonary tissues. Western blotting demonstrated that the protein expression levels of p-IκB, p-P65, and P65 in rat pulmonary tissues of PAH group were remarkably higher than CTL group (p<0.01). However, they were notably down-regulated in PAH + sevoflurane group when compared with those in PAH group (p<0.05). The above experimental results suggested that the NF-κB signaling pathway in pulmonary tissues of rats in PAH group was activated and was inhibited by sevoflurane. Subsequent RT-qPCR results indicated that no significant (N.S.) differences were observed in the mRNA level of P65 among the three groups. Compared with CTL group, PAH group showed significantly up-regulated levels of Ki-67 and α-SMA in rat pulmonary tissues (p<0.01). However, their expression levels were markedly reduced in PAH + sevoflurane group when compared with PAH group (p<0.05). Finally, the detection of pulmonary circulatory function-related indicators illustrated that RVSP and RVHI increased significantly in PAH group in comparison with CTL group. However, they declined remarkably in PAH + sevoflurane group when compared with those in PAH group (p<0.05). CONCLUSIONS: Sevoflurane down-regulates the levels of p-IκB, p-P65, and P65 to repress the activation of the NF-κB signaling pathway. This may reduce pulmonary fibrosis and ultimately prevent PAH.

lncRNA-RMRP promotes proliferation, migration and invasion of bladder cancer via miR-206.

OBJECTIVE: The incidence of bladder cancer (BC) is common in the world, but its detail mechanisms for occurrence and development remain unclear. Recently, long non-coding RNAs (lncRNAs) have been observed to play an important role in many different diseases. In this research, we mainly explored the role of the RNA component of mitochondrial RNA processing endoribonuclease (lncRNA-RMRP) in bladder cancer. MATERIALS AND METHODS: We used qRT-PCR to detect the expression of lncRNA-RMRP in bladder cancer patients and tumor cells, and the clinical significance was also analyzed. The methyl thiazolyl tetrazolium (MTT) assay was used to detect the cell proliferation, and we used transwell to detect the migration and invasion, after the lncRNA RMRP was inhibited. Western-blot was used to measure the relative protein expression level in bladder cancer cells after transfection with siRNA-NC or siRNA-RMRP. RESULTS: We found that the lncRNA RMRP was highly expressed in bladder cancer tissue, compared with adjacent tissue. We also found that the expression of RMRP was closely related with the size, lymph node metastasis and survival time of patients. What's more, RMRP could promote the proliferation, migration and invasion of BC cell lines via regulating miR-206 as a sponge. CONCLUSIONS: According to the results, we found that lncRNA RMRP was closely related to the progression of bladder cancer, which could be a potential target for treating BC patients.

[Treatment principle and surgical technique of pulmonary ground glass nodules].

Pulmonary ground glass nodule (GGN) is a term of radiological manifestation, which may be malignant or benign. GGN's imaging performance is diverse, and the management for pulmonary GGN remains controversial. Numerous clinical studies have clarified the safety of GGN follow-up and longer follow-up intervals, stricter surgical or biopsy indications are recommended. In clinical practice, the size of GGN, the size of consolidation, dynamic change during follow-up are the factors that help surgeons to decide the timing of surgery. There are some misunderstandings for the management of GGN, such as the administration of antibiotics, the use of PET-CT, pure GGN adjacent to visceral pleura, and GGN with penetrating vessel. Segmentectomy for ground glass nodules is being accepted by more and more surgeons. Through theoretical study and clinical practice, surgeons can master anatomical segmentectomy.

Development of a peptide ELISA to discriminate vaccine-induced immunity from natural infection of hepatitis A virus in a phase IV study.

Hepatitis A virus (HAV) is a highly infectious agent that causes acute liver disease. The infection can trigger the production of antibodies against the structural and non-structural proteins of HAV. Nonetheless, vaccination with an HAV vaccine leads to the production of a primary antibody against the structural proteins. Because the non-structural proteins are only produced during active virus replication, there is no or very little antibody production against the non-structural proteins. However, the current commercial immunoassay cannot distinguish between antibodies produced during natural infection and those from vaccination against HAV. In our study, six immune-dominant epitopes from the non-structural proteins were designed, synthesized, linked together and cloned into pGEX-5X-1 plasmid. The recombinant protein was expressed in E. coli and purified by Ni2+-coated magnetic agarose beads. Then the purified recombinant protein was used as an ELISA antigen to detect antibodies for HAV non-structural proteins in serum samples. Seventy-seven attenuated and 89 inactivated vaccinated samples collected from our previous phase IV study of HAV vaccines were detected by peptide ELISA developed in this study. The mean OD450 value for the vaccination samples and acute infection samples were 0.529 (0.486 for the attenuated group and 0.567 for the inactivated group) and 1.187, respectively. According to the receiver operating characteristic (ROC) curve, the sensitivity and specificity of the peptide ELISA were 93.80% and 91.00%, respectively. This peptide ELISA was confirmed to discriminate vaccine-induced immunity from natural infection of HAV in a phase IV study with high sensitivity and specificity.

Transcription factor aryl hydrocarbon receptor/aryl hydrocarbon receptor nuclear translocator is involved in regulation of the xenobiotic tolerance-related cytochrome P450 CYP6DA2 in Aphis gossypii Glover.

The cotton aphid, Aphis gossypii, is one of the most economically important agricultural pests worldwide as it is polyphagous and resistant to many classes of insecticides. Overexpression of the cytochrome P450 monooxygenase (P450) CYP6DA2 has previously been found to be associated with gossypol and spirotetramat tolerance in the cotton aphid. In the present study, the elements located in the promoter region (-357:-343; -250:-241; -113:-104) of CYP6DA2 were shown to control promoter activity, and gossypol induction was observed. We hypothesized that the expression of CYP6DA2 is subject to transcriptional regulation. To investigate the underlying mechanism, we assessed two transcription factors, aryl hydrocarbon receptor (AhR) and aryl hydrocarbon receptor nuclear translocator (ARNT), and found that the abundance of AhR was highly correlated with CYP6DA2 abundance. RNA interference of AhR or ARNT significantly decreased the levels of the target gene as well as those of its counterpart, and both dramatically repressed CYP6DA2 expression. Cotransfection of the ARNT, AhR, or AhR plus ARNT and CYP6DA2 promoter constructs elevated CYP6DA2 promoter activity, with the AhR plus ARNT cotransfection being the most effective. Thus, these elements located in the promoter were responsible for CYP6DA2 transcription, and CYP6DA2 expression was regulated by the transcription factors AhR and ARNT.

A rice jacalin-related mannose-binding lectin gene, OsJRL, enhances Escherichia coli viability under high salinity stress and improves salinity tolerance of rice.

Salinity, which is one of the most common abiotic stresses, may severely affect plant productivity and quality. Although plant lectins are thought to play important roles in plant defense signaling during pathogen attack, little is known about the contribution of plant lectins to stress resistance. We cloned and functionally characterized a rice jacalin-related mannose-binding lectin gene, OsJRL, from rice 'Nipponbare'. We analyzed the expression patterns of OsJRL under various stress conditions in rice. Furthermore, we overexpressed OsJRL in Escherichia coli and rice. The cDNA of OsJRL contained a 438 bp open reading frame, which encodes a polypeptide of 145 amino acids. OsJRL was localized in the nucleus and cytoplasm. Real time PCR analyses revealed that OsJRL expression showed tissue specificity in rice and was upregulated under diverse stresses, namely salt, drought, cold, heat and abscisic acid treatments. Overexpression of OsJRL in E. coli enhanced cell viability and dramatically improved tolerance of high salinity. Overexpression of OsJRL in rice also enhanced salinity tolerance and increased the expression levels of a number of stress-related genes, including three LEA (late embryogenesis abundant proteins) genes (OsLEA19a, OsLEA23 and OsLEA24), three Na+ transporter genes (OsHKT1;3, OsHKT1;4 and OsHKT1;5) and two DREB genes (OsDREB1A and OsDREB2B). Based on these results, we suggest that OsJRL plays an important role in cell protection and stress signal transduction.

Landscape of expression profiles in esophageal carcinoma by The Cancer Genome Atlas data.

In this study, we explored the gene and microRNA (miRNA) expressions profile of esophageal carcinoma. The expression data for messenger RNAs and miRNAs in normal and cancerous esophageal tissues were obtained from the Cancer Genome Atlas database and then the differentially expressed genes and miRNAs were identified. As a result, we identified 2962 genes and 45 miRNAs differentially expressed in esophageal carcinoma compared with normal esophageal tissues. Subsequently, the altered gene functions and signaling pathways were investigated using gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, and these differentially expressed genes were significantly enriched in the cell cycle, cell migration, mitogen-activated protein kinase (MAPK) and toll-like receptor signaling pathway, and so on. Then the regulatory relationships between the differentially expressed miRNAs and genes were examined with Targetscan and Miranda, and the potential target sites of transcription factors (TFs) in the promoter regions of these miRNAs and genes were identified using the TRANSFAC database. Finally the TF-miRNA-gene network in esophageal cancer was established, summarizing the regulatory links among the TFs, differentially expressed miRNAs and differentially expressed genes. Factors such as core promoter-binding protein (CPBP), nuclear factor of activated T-cells 1 (NFAT-1), miR-30c-5p, were located in the central hub of this network, highlighting their vital roles in esophageal tumorigenesis. These findings may extend our understanding of the molecular mechanisms underlying esophageal carcinoma and promote new perspectives for prevention, diagnosis and treatment.

Construction of a natural phage antibody library of human anaplastic thyroid carcinoma.

The objective of this study was to identify and construct a human natural phage single-chain antibody (scFv) library of human anaplastic thyroid carcinoma (ATC) using phage display technology. Total RNA was extracted from lymphatic tissue near an ATC and used to amplify variable heavy chain (VH) and variable light chain (VL) fragments with added linker sequences using reverse transcription-polymerase chain reaction (RT-PCR). After purification, the VH and VL amplicons were used to produce scFv fragments with added SfiI and NotI restriction enzyme recognition sites using splicing-overlap-extension PCR. Following digestion, the scFv gene was cloned in the pCANTAB-5E plasmid, and the recombinant phagemids were transformed into the susceptible Escherichia coli TG1 strain. After infection by the helper phage M13K07, a human ATC phage antibody library was successfully constructed. Clear 28 S and 18 S bands could be seen in the total RNA from the library, and the sizes of the VH, VL, and scFv genes contained therein were approximately 370, 350, and 750 bp, respectively. In addition, the conversion efficiency as measured by the pUC19 standard plasmid was 10(8) CFU/µg, and the positive insert ratio was 86.4% (19/22). These results demonstrated the successful construction of a human ATC scFv antibody gene library, and might provide the experimental basis for the further screening and identification of a phage single-chain antibody with ATC cell-specificity.

Circulating miR-125b as a biomarker of Ewing's sarcoma in Chinese children.

Previous studies indicated that microRNA-125b (miR-125b) has an important role in the progression of Ewing's sarcoma (ES). The purpose of the current study was to examine expression changes of miR-125b in the serum of ES patients and evaluate if the expression level of miR-125b could serve as a new biomarker for ES. This study was performed on patients who underwent surgical resection at our hospital between 2005 and 2013 after an initial diagnosis of ES. We measured serum miR-125b levels in 63 patients with ES and 126 healthy control patients using a real-time quantitative reverse transcriptase-PCR (qRT-PCR) method. Expression levels of serum miR-125b were distinctly decreased in ES patients when compared with healthy controls (P < 0.001). ES cases that had a poor response to chemotherapy presented a significant down-regulation of miR-125b (P = 0.001). The ROC curve showed that the serum miR-125b could serve as a valuable biomarker for differentiating ES patients from healthy controls with an AUC of 0.879 (95%CI = 0.817-0.924; P < 0.001). At a cut-off value of 2.203 for miR-125b, the sensitivity was 72.8% and the specificity was 87.2% in discriminating ES from the controls. Our results indicate that serum miR- 125b may serve as a useful noninvasive biomarker for ES.

Ropivacaine via trans-cricothyroid membrane injection inhibits the extubation response in patients undergoing surgery for maxillary and mandibular fractures.

Extubation response can lead to cardiovascular and respiratory complications. Here, we aimed to evaluate the effect of ropivacaine injected via the trans-cricothyroid membrane on the extubation response. This prospective, double-blind, randomized study included 70 patients classified as American Society of Anesthesiologists status I-II, who required general anesthesia with nasotracheal intubation for maxillary and mandibular fracture surgery; patients were divided into the ropivacaine (20 mg) and dicaine (20 mg) groups. Both groups were injected via the trans-cricothyroid membrane. Mean arterial pressure (MAP), heart rate (HR), and incidence and severity of cough were recorded during intubation and extubation. During intubation, there was no significant intergroup difference in MAP or HR and no occurrence of coughing (P > 0.05). During extubation, MAP and HR were significantly lower in the ropivacaine group than the dicaine group (P < 0.05). The proportion of patients with no reports of cough was significantly higher in the ropivacaine group than in dicaine group (P < 0.05). The number of patients with grade 1 or 2 cough was significantly higher in the dicaine group than that in the ropivacaine group (P < 0.05). There was no significant intergroup difference in the rate of postoperative complications (P > 0.05). These results suggest that the administration of ropivacaine via trans-cricothyroid membrane injection can effectively inhibit the extubation response.