Objective To observe the effects of microRNA-21 (miR-21) inhibitor on apoptosis of type Ⅱalveolar epithelial cells (AEC Ⅱ) in rats with hyperoxia-induced acute lung injury (HALI).Methods Eighty Sprague-Dawley (SD) rats were divided into air-control group,hyperoxia injury group,empty-virus control group (200 μL solution with lentivirus was dropped into the nasal) and miR-21 inhibitor pretreatment group (200 μL solution with lentivirus contained miR-21 inhibitor was dropped through the nasal) by random number table.After treatment,the rats in all groups were fed in the hyperoxia incubator with oxygen concentration exceeding 90％ for production of HALI model,and the rats in air-control group were fed normally without any treatment.Ten rats were selected at 0,24,48 and 72 hours after exposure in hyperoxia environment respectively,and the general changes of lung tissues were observed in light microscope.The right lung tissues were harvested to observe the pathological changes under light microscopy.The left lung tissues of other 10 rats in each group were harvested at 48 hours after execution,the miR-21 expression was determined by real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR),the protein expression of cysteinyl aspartate-specific proteinase-3 (caspase-3) was determined by Western Bolt,and apoptosis of AEC Ⅱ was detected by TdT-mediated dUTP nick end labeling (TUNEL).Results ① No abnormal appearance in lung tissues was observed at all time points in the air-control group.In hyperoxia injury group,the lung injury would be more severe if the exposure time was longer,and lung tissues turned dark red after exposure for 72 hours,with patchy hemorrhage in several places;the structure of lung tissues was disordered,the alveolar wall was broken,the alveolar septum was significantly edematous and broadened,and there was plenty of inflammatory cell infiltration and edema fluid appeared inside the alveolar space.In miR-21 inhibitor pretreatment group,the degree of lung tissue injury was more severe than that of the hyperoxia injury group,and there was no significant change in empty-virus control group.(②) Compared with air-control group,miR-21 expression of the hyperoxia injury group was significantly decreased (2-△△Ct:0.021 ± 0.005 vs.0.037 ± 0.006),and the protein expression of caspase-3 was significantly increased (A value:0.423±0.081 vs.0.123±0.023,both P ＜ 0.05).After pretreatment with miR-21 inhibitor,the expression of miR-21 was further decreased (2-△△Ct:0.014±0.003 vs.0.021 ±0.005),while the protein expression of caspase-3was further increased (A value:0.691 ±0.085 vs.0.423 ±0.081,both P ＜ 0.05).There were no statistically significant differences in the expression of miR-21 (2-△ △ct:0.025 ± 0.007 vs.0.021 ± 0.005) and caspase-3 (A value:0.475 ± 0.062vs.0.423 ±0.081) between empty-virus control group and hyperoxia injury group (both P ＞ 0.05).(③) Compared with air-control group,the apoptosis cells in hyperoxia injury group were increased,which was further increased after pretreatment of miR-21 inhibitor,but no changes were found in empty-virus control group.Conclusion Inhibition of miR-21 expression in vivo could aggravate the injury of lung tissue in HALI rats,and increase the apoptosis of AEC Ⅱ.

Objective:To explore whether microRNA-21-5p (miR-21-5p) has the effect of anti-apoptosis of human alveolar typeⅡ epithelial cells (ATⅡ).Methods:ATⅡ cells derived from the human were cultured in vitro and used for experiments when the cells were grown until the presence of lamellar bodies and microvilli were observed by light microscope. The cells were divided into blank control group (direct culture), hydrogen peroxide (H 2O 2) injury group (cultured with 0.5 mmol/L H 2O 2), and miR-21-5p overexpression group (using miR-21-5p with a multiplicity of infection (MOI) of 100 lentiviral overexpression vector with 0.5 mmol/L H 2O 2) and miR-21-5p empty virus control group (miR-21-5p lentiviral blank vector was co-cultured with 0.5 mmol/L H 2O 2). In each group, cell proliferation was detected by cell counting kit-8 (CCK-8) at 0, 12, 24, 36, and 48 hours of cell culture; cell apoptosis was detected by flow cytometry at 24 hours of culture. Results:① Cell proliferation activity test results: with the extension of cell culture time, the cell proliferation activity of the blank control group gradually increased, while the cell proliferation activity gradually decreased after the addition of 0.5 mmol/L H 2O 2. However, the cells proliferation activity in the miR-21-5p overexpression group decreased more slowly than that in the H 2O 2 injury group and the miR-21-5p empty virus control group, and the cell proliferation activity at 48 hours was significantly higher than the H 2O 2 injury group and the miR-21-5pempty virus control group ( A value: 0.295±0.005 vs. 0.184±0.005, 0.169±0.002, both P < 0.05). It showed that both H 2O 2 and lentivirus accelerated cell damage, while miR-21-5p could reduce cell apoptosis. ② Apoptosis rate test results: compared with the blank control group, the apoptosis rate increased significantly after adding 0.5 mmol/L H 2O 2; while the apoptosis rate of the miR-21-5p overexpression group was lower than that of the H 2O 2 injury group and miR-21-5p empty virus control group [early apoptosis rate: (14.31±0.12)% vs. (24.50±0.12)%, (23.41±0.13)%; late apoptosis rate: (8.12±0.13)% vs. (9.71±0.11)%, (10.41±0.15)%; overall apoptosis rate: (22.33±0.12)% vs. (34.21±0.10)%, (33.82±0.14)%; all P < 0.05], which further proved that miR-21-5p had anti-apoptotic effects. Conclusion:miR-21-5p has an anti-apoptotic effect on human ATⅡ.

?AIM:To compare the changes of the contrast sensitivity after LASIK with femtosecond laser and microkeratome and to explore the influence of different methods making corneal flap on visual quality.? METHODS: There were 212 eyes in 106 myopes underwent excimer operation . According to the different methods of operation, they were divided into two groups:microkeratome group ( SBK group ) and femtosecond laser group ( FS group) . FS group: a total of 112 eyes in 56 patients received LASIK with femtosecond laser. SBK group: a total of 100 eyes in 50 patients received LASIK with microkeratome. Contrast sensitivity was detected preoperatively, and 1wk, 3mo postoperatively and compared between the two groups.? RESULTS: At 1wk after operation, the contrast sensitivity under photopic environment decreased in the two groups, compared with those before operation ( P<0. 05). The differences of contrast sensitivity before and 3mo after operation were not significant (P>0. 05). No statistical significant difference was found in contrast sensitivity under photopic environment at 1wk, 3mo between the two groups ( P>0. 05 ). At 1wk after the operation, the contrast sensitivity under scotopic environment decreased in both groups compared with those before operation ( P< 0. 05 ). In SBK group, it decreased more than in FS group (P<0. 05). After 3mo, the decline of 14. 2c/d spatial frequency contrast sensitivity under scotopic environment in the SBK group was more than other frequency. No statistical significant difference was found in the rest frequency contrast sensitivity under scotopic environment before and after operation (P>0. 05). After 1wk, contrast sensitivity with glare stimulation in both groups decreased, compared with those before operation ( P< 0. 05 ), while in SBK group, it decreased more than in FS group (P<0. 05). After 3mo, except that the decline of 14. 2c/d spatial frequency contrast sensitivity with glare stimulation in the SBK group was significant compared with those before operation, the contrast sensitivity under glare stimulation in both groups had no significant differences compared with before operation(P>0. 05).?CONCLUSION:LASIK with femtosecond laser can get a better visual quality than LASIK with microkeratome.

OBJECTIVETo investigate the optimal dosage and timing for 5-bromodeoxyuridine (BrdU) labeling of endothelial progenitor cells (EPCs) from rat circulating blood.

METHODSThe animal model for rat tooth movement was established. EPCs were obtained by density gradient centrifugation. The expressions of specific antigens on cell surface were analysed by immunocytochemistry and fluorescenceochemistry. EPCs were incubated with BrdU at different concentrations (5, 10, 15 micromol/L) for different incubating time (12, 24, 48, 72, 96 h) to identify the optimal BrdU concentration and incubating time for cell labeling. Immunohistochemistry was performed to calculate the labeling index (LI).

RESULTSThe culture cell positively expressed CD34, CD133 and could be shown to endocytose DiI-ac-LDL, FITC-UEA-1. Incubation of the EPCs with BrdU at 10 micromol/L and for an optimal length of 72 h appeared to achieve the highest LI (66.8+/-2.9)%, which was significantly higher than group of 5 micromol/L (P<0.05), while there was no significant difference between the group of 15 micromol/L and 10 micromol/L (P>0.05).

CONCLUSIONEPCs can be isolated from tooth movement rat circulating blood and cultured. Incubation of the EPCs with BrdU at 10 micromol/L and for an optimal length of 72 h appeared to achieve the optimal LI. This provides a foundation for us to investigate the mechanism of chemiotaxis and differentiation for EPCs.

Animals ; Bromodeoxyuridine ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Endothelial Cells ; Endothelial Progenitor Cells ; Rats ; Stem Cells ; Tooth Movement Techniques

S100A8, an important member of the S100 protein family, is a low-molecular-weight (10.8 kDa) calcium-binding protein containing conserved EF-hand structural motifs. Previous studies have shown that the biological function of S100A8 protein is associated with a variety of inflammatory diseases, for example asthma. S100A8 protein plays important roles in the regulation of inflammation. It can activate inflammatory cells and cytokines via chemotactic activity for neutrophils, and bind to the receptor for advanced glycation end products (RAGE) and Toll-like receptor 4 (TLR4), thus mediating intracellular inflammatory signaling transduction. Additionally, recent studies have reported the anti-inflammation activity of S100A8 protein, which indicates that S100A8 may have a more complex function of biological regulation in the different pathophysiological conditions. In this review, we summarized the studies on the functions and molecular mechanisms of S100A8 protein in inflammation, which would propose a novel strategy for the prophylaxis and treatment of asthma and other inflammatory diseases.
Animals ; Asthma ; physiopathology ; Calgranulin A ; physiology ; Humans ; Inflammation ; physiopathology

OBJECTIVETo explore the role of Xuebijing Injection (XBJI) in inhibiting inflammatory factors associated with anoxia/reoxygenation myocardial inflammatory response of rats.

METHODSTotally 36 healthy male Sprague-Dawley rats, 280 ± 30 g were randomly divided into six groups, i.e., the normal control group (N group), the balanced perfusion group (BP group),the model group (M group),the low dose XBJI group (XBJI(L) group), the middle dose XBJI group (XBJI(M) group),and the high dose XBJI group (XBJI(H) group), 6 in each group. The myocardial anoxia/reoxygenation rat model was established by Langendorff isolated heart perfusion. The concentration of TNF-α in the myocardial tissue was detected by ELISA. The expression of nuclear factor kappa B p65 (NF-κB p65) protein and Toll like receptor 4 (TLR4) protein were detected using Western blot. The expression of NF-κB p65 mRNA and TLR4 mRNA was detected by RT-PCR. Ultrastructural changes of anoxia-reoxygenation rats' heart muscle were observed under transmission electron microscope.

RESULTSCompared with the M group,the TNF-α concentration, expression levels of NF-κB p65 protein and mRNA, TLR4 protein and mRNA decreased to various degrees in the XBJI(L) group, the XBJI(M) group, and the XBJI(H) group. The TNF-α expression level decreased most significantly in the XBJI(L), group (P < 0.01), while other indices decreased most obviously in the XBJI(M) group (P < 0.01, P < 0.05). Expression levels of NF-κB p65 and TLR4 protein were obviously lower in the XBJI(M) group than in the XBJI(L) group (P < 0.05). There was no statistical difference in other indices among the three XBJI groups (P > 0.05). Myocardial fibers were loose and broken with disappearance of transverse striation, and mitochondrial cristae was dissolved and severely damaged in the M group. The aforesaid condition was improved after treated by XBJI, with the most obvious effect obtained in the XBJI(M) group.

CONCLUSIONSDifferent doses of XBJI could attenuate inflammatory reactions after myocardial anoxia/reoxygenation rats' heart muscle through inhibiting TLR4-NF-κB-TNF-α signal transduction pathway. The best effect could be obtained by 4 mL/100 mL XBJI.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Hypoxia ; Male ; Myocardium ; metabolism ; Myocytes, Cardiac ; NF-kappa B ; metabolism ; Oxygen ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Toll-Like Receptor 4 ; metabolism ; Transcription Factor RelA ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo systematically review the effectiveness and safety of open and laparoscopic surgeries for treatment of adrenal tumors.

METHODSThe online databases including CNKI, PUBMED, SinoMed, EBSCO, Springerlink, WanFang Data, and VIP were searched for clinical trials published from 1999 to 2016. A meta-analysis was performed using RevMan 5.2 software.

RESULTSA total of 2340 patients in 25 trials were included. The results of meta-analysis showed that laparoscopic surgery was better than open surgery in terms of intestinal function recovery time (OR=-0.96, 95%CI [-1.22, -0.70] P<0.000 01), hospitalization time (OR=-3.48, 95%CI [-4.13, -2.78], P<0.000 01), complications (OR=0.22, 95%CI [0.14, 0.35], P<0.0001), and volume of blood loss (OR=-104.77, 95%CI [-138.95, -70.60], P<0.000 01). There was no significant difference in the surgery cost between open and laparoscopic surgeries.

CONCLUSIONLaparoscopic surgery is superior to open surgery for treatment of adrenal tumors for shorter intestinal function recovery time, surgery duration, and hospitalization time and less complications and blood loss.

OBJECTIVETo study the methods of decalcification for making united slices of tooth and affiliated periodontic tissues.

METHODSTwenty-one samples containing dog molars and affiliated periodontic tissues were divided into seven mean groups. The pH value of solution, time of decalcification, weight and volume of samples, and content of decalcified calcium were detected. The slices were observed by HE, specific, and immunohistochemical stain.

RESULTSThe velocity of decalcification increased with decrease of solution pH. The weight of samples lightened by 37.61%, the volume reduced by 25.97% on average, and calcium decalcified was 174.49 mg per gram humid samples. The EDTA decalcification was slowest, but it was best. Decalcification was fast in Plank-Rycho solution while the section was worst, and faster in the formyl solution containing aluminium chloride than in EDTA, and the section was better.

CONCLUSIONSThe 50% formyl solution containing aluminium chloride is an ideal decalcifying solution.

Animals ; Decalcification Technique ; methods ; Dogs ; Edetic Acid ; Formates ; Microtomy ; Molar ; Periodontium

Objective:To investigate the prognostic factors of patients with esophageal squamous cell carcinoma with pulmonary metastasis.Methods:Clinical characteristics of 135 esophageal squamous cell carcinoma patients presenting with pulmonary metastasis after treatment in Zhejiang Cancer Hospital from 2008 to 2018 were retrospectively analyzed. Thesurvival rate was calculated by Kaplan-Meier method. Univariate analysis was performed by log-rank test. Multivariate prognostic analysis was conducted by Cox models.Results:The median follow-up time of 135 patients with esophageal squamous cell carcinoma was 94.2 months (19.5-258.9 months), and 109 patients died (80.7%). The 1-and 2-year overall survival rates were 47.4% and 25.1%, with the median survival time was 11.1 months (7.3-14.9 months). Univariate prognostic analysis showed that age, number of lung metastases, treatment of lung metastases, lymph node metastasis, distant organ metastasis, and the interval between the first treatment and lung metastasis were the prognostic factors of esophageal squamous cell carcinoma with lung metastasis (all P<0.05). Multivariate analysis demonstrated that age and number of lung metastases were the independent prognostic factors for patients with esophageal squamous cell carcinoma with lung metastases (all P<0.05). Conclusions:Age and number of lung metastases are the independent prognostic factors for patients with esophageal squamous cell carcinoma with lung metastases. Surgery or radiotherapy-based regional therapy can enhance clinical prognosis.

Objective To establish a druggability evaluation method for new targets of anti-tumor drugs by analyzing the mutation genes of common tumors in the digestive system. Methods We collected the mutant gene data of the five common tumors of the digestive system (esophageal cancer, gastric cancer, colorectal cancer, liver cancer and pancreatic cancer) in the Integrative Onco Genomics database, and screened out the genes with higher mutation rates in each tumor. We evaluated the druggability of these genes or their encoded proteins, and discovered the potential targets for the new anti-tumor drugs. Results A total of five tumors, 35 cohorts and 5445 tumor samples were collected in this study. The top 10 mutation genes were selected for further analysis. The canSAR database was used to analyze the druggability of unpublished mutant genes or their encoded proteins, and a total of 17 potential therapeutic drug targets were screened out. Conclusion A method for evaluating druggability of targets based on mutant genes or their encoded protein is established in this study. The application of this method can provide a reference for discovering new anti-tumor therapeutic target, saving the cost and time of target screening in new drug development.