BACKGROUND:Circular RNAs(circRNAs)play important roles in a variety of diseases or tumors,and recent findings have revealed that circRNAs are abnormally expressed in knee osteoarthritis and are involved in disease progression through microRNA/mRNA regulation. OBJECTIVE:To investigate the effect of circRNA WD repeat containing protein 1(circ-BRWD1)/miR-488-3p/DNA methyltransferase 3A(DNMT3A)on the proliferation and apoptosis of chondrocytes in knee osteoarthritis. METHODS:Real-time fluorescence quantitative PCR was performed to detect the expression of circ-BRWD1,miR-488-3p,DNMT3A in knee osteoarthritis chondrocytes.Cells were divided into si-NC group,si-circ-BRWD1 group,vector group,circ-BRWD1 group,si-circ-BRWD1+anti-miR-NC group,si-circ-BRWD1+anti-miR-488-3p group,miR-NC group,miR-488-3p group,anti-miR-NC group,anti-miR-488-3p group,miR-488-3p+vector group,miR 488-3p+DNMT3A group.Real-time fluorescence quantitative PCR was used to detect circ-BRWD1,miR-488-3p,DNMT3A expression,MTT and flow cytometry assay were used to detect cell proliferation and apoptosis.Western blot assay was used to detect DNMT3A and proliferation/apoptosis-related protein expression.Dual luciferase reporter assay was used to Dual luciferase reporter assay to detect the targeting relationship of circ-BRWD1 with miR-488-3p and miR-488-3p with DNMT3A. RESULTS AND CONCLUSION:circ-BRWD1 and DNMT3A were highly expressed and miR-488-3p was lowly expressed in knee osteoarthritis chondrocytes compared with normal chondrocytes.Knockdown of circ-BRWD1 or overexpression of miR-488-3p inhibited proliferation and induced apoptosis in knee osteoarthritis chondrocytes.circ-BRWD1 targeted negative regulation of miR-488-3p and inhibition of miR-488-3p reversed the effect of circ-BRWD1 knockdown on chondrocyte proliferation and apoptosis in knee osteoarthritis.miR-488-3p targeted negative regulation of DNMT3A and upregulation of DNMT3A reversed the effect of miR-488-3p overexpression on chondrocyte proliferation and apoptosis in knee osteoarthritis.circ-BRWD1 could regulate the expression of DNMT3A by regulating miR-488-3p.To conclude,knockdown of circ-BRWD1 inhibits chondrocyte proliferation and induces apoptosis in knee osteoarthritis,and the mechanism of action may be related to the regulation of miR-488-3p/DNMT3A axis.

Circular RNAs (circRNAs) are a new class of non-coding RNAs, which have been confirmed to regulate insect gene expression and immune response through multiple manners such as competing endogenous RNA (ceRNA) regulatory network. Currently, function of circRNA in honey bee immune response remains unclear. In this study, PCR and Sanger sequencing were performed to validate the back splicing (BS) site of ame_circ_000115 (in short ac115). RT-qPCR was used to detect the expression profile of ac115 in larval guts of Apis mellifera ligustica stressed by Ascosphaera apis. Dual-luciferase reporter gene assay was conducted to verify the binding relationship between ac115 and ame-miR-13b. Interference of ac115 in larval guts was carried out by feeding specific siRNA, followed by determination of the effect of ac115 interference on expression of six genes relevant to host immune response. The results confirmed the existence of BS site within ac115. Compared with the un-inoculated group, the expression of ac115 in 4-day-old larval gut of the A. apis-inoculated group was up-regulated with extreme significance (P < 0.000 1), while that in 5- and 6-day-old larval guts were significantly up-regulated (P < 0.05). The brightness of specific band for ac115 in 4-, 5- and 6-day-old larval guts of the siRNA-circ_000115-fed group gradually became weak, whereas that of the siRNA-scrambl-fed group was pretty high without obvious variation. Compared with that of the siRNA-scramble-fed group, the expression of ac115 in 4-day-old larval gut of the siRNA-circ_000115-fed group was significantly down-regulated (P < 0.05), whereas that of the 5- and 6-day-old larval guts were down-regulated with extreme significance (P < 0.001). Ame-miR-13b was truly existed and expressed in A. m. ligustica larval guts, and there was true binding relationship between ac115 and ame-miR-13b. Compared with that of the siRNA-scramble-fed group, the expression of antimicrobial peptide genes hymenoptaecin and abaecin in 6-day-old larval gut of the siRNA-circ_000115-fed group was significantly up-regulated (P < 0.05), while that of ecdysone receptor (Ecr) was down-regulated with extreme significance (P < 0.01). These results indicate that ac115 is truly expressed in A. m. ligustica larval guts, BS site truly exists within ac115, and effective interference of ac115 in A. m. ligustica larval guts can be achieved via feeding siRNA. Moreover, ac115 potentially regulates Ecr expression through adsorption of ame-miR-13b and expression of hymenoptaecin and abaecin using a non-ceRNA manner, further participating in host stress-response.
Bees/genetics* ; Animals ; Larva/metabolism* ; RNA, Circular/genetics* ; RNA, Small Interfering/genetics* ; MicroRNAs/genetics*

Objective:To investigate the clinical manifestations, auxiliary examination results, prognosis and treatment of children with typical hemolytic uremic syndrome (D + HUS). Methods:The clinical data of 36 patients diagnosed as D + HUS in the Department of Pediatrics of Nanjing Jinling Hospital from January 2001 to January 2019 were collected, and the laboratory results including blood routine, liver and kidney function, coagulation function, humoral immunity and urine were compared before and after treatment. Results:The white blood cell count[ (9.28±6.77)×10 9/L vs.(11.20±5.93) ×10 9/ L ], C-reactive protein [7.15(3.34, 29.33) mg/L vs.31.83(25.03, 39.75) mg/L], reticulocyte count [(112.49±76.25)×10 9/L vs. (206.49±147.99)×10 9/L], erythrocyte sedimentation[15.02(11.79, 22.83) mm/1 h vs.28.06(24.13, 40.52) mm/1 h] , aspartate aminotransferase[50.04(41.92, 60.11) U/L vs.62.61(54.58, 83.52) U/L], alanine aminotransferase [16.72(11.80, 24.74) U/L vs.24.54(20.30, 34.36) U/L], uric acid [(532.84±309.06) μmol/L vs.(606.64±327.23) μmol/L], serum creatinine[160.07(124.87, 221.18) μmol/L vs.200.56(160.62, 283.01)μmol/L ], blood urea nitrogen [20.74(15.77, 28.40) mmol/L vs.33.67(25.91, 45.84) mmol/L], lactate dehydrogenase [488.21(337.59, 692.82) U/L vs.1 520.68(734.24, 2 272.10) U/L ], prothrombin time [(12.14±5.89) s vs. (17.91±6.12) s ], activated partial thrombin time [(25.05±6.26) s vs.(32.38±5.49) s], fibrinogen [ (3.79±2.17) g/L vs.(5.17±3.88) g/L], D-dimer [0.92(0.30, 1.13) mg/L vs. 1.27(1.01, 1.90) mg/L ], 24-hour urinary proteinuria [ (84.05±44.19) mg/(kg·24 h) vs.(112.18±78.26) mg/(kg·24 h) ], urinary sediment [175.73(79.72, 258.66)×10 7/L vs. 160.38(118.68, 361.83)×10 7/L], N-acetyl-β-D-glucosaminidase [25.10(18.84, 33.02) U/(g·cr) vs. 41.57(29.49, 58.61) U/(g·cr)], urinary retinol binding protein [0.35(0.18, 1.33) mg/L vs 1.05(0.66, 1.68) mg/L.] in patients after treatment were significantly lower than those before treatment, and the differences were all statistically significant(all P<0.05); patients had higher levels of red blood cell count [ (4.51±1.73)×10 9/L vs.(2.43±1.40) ×10 9/L], platelet[(126.82±78.35)×10 9/L vs. (85.21±69.38)×10 9/L], hemoglobin[(118.46±18.27) g/L vs. (62.36±16.11) g/L], and complement C 3levels [(0.74±0.39) g/L vs.(0.58±0.27) g/L ] after treatment, and the differences were all all statistically significant(all P<0.05). Children with D + HUS showed multiple system injuries.Among 36 cases, 17 cases (47.22%) had fever, 31 cases (86.11%) had abdominal pain and diarrhea, 29 cases (80.56%) had nausea and vomiting, 8 cases (22.22%) had headache and dizziness, 36 cases (100.00%) had proteinuria and hematuria, 34 cases (94.44%) had renal insufficiency, and 21 cases (58.33%) had yellow staining of skin and sclera.The auxiliary examination for abnormal results mainly included renal pathology (100.00%) (mesangial proliferation endothelial cell proliferation and swelling, and shedding of renal tubular brush borders), bone marrow pathology (100.00%) (active bone marrow hyperplasia), and renal B-ultrasound (86.67 %) (kidney injury-like sound image). Conclusions:D + HUS in children shows multiple system damage.Digestive system abnormalities are the main causative factor of D + HUS in children, and the disease is dangerous.Therefore, early diagnosis and active treatment can improve the prognosis.

Objective To detect the consistency of EGFR mutations in pleural effusion in terminal non-small cell lung cancer(NSCLC)patients by amplification refractory mutation system- polymeric chain reaction (ARMS-PCR).Methods Pleural effusion from 44 cases with advanced NSCLC were collected.DNA was extract-ed from a part of pleural effusion,and EGFR gene18,19,20,and 2 l exons mutations were detected by ARMS-PCR amplification. The left part of pleural effusion was used for the pathological microscopic examination of the embedding and to evaluate the feasibility of pleural effusion direct detection.Part of the case results were compared with tissue or pleural effusion at the same time. Results The mutation rate of EGFR detection in pleural effusion was 54.5%,including 19-del(50.0%)and 21-L858R(50.0%);75.0% of pleural effusion sediment was morpho-logically diagnosed with adenocarcinoma and EGFR mutation rate was 27.3% in pleural effusion with no tumor cell. The consistent rate was 72.7% in 22 cases of pleural effusion and tissue and that was 50.0% in 16 cases of pleural effusion and plasma.The comparison between pleural effusion and organizations and plasma test results was statisti-cally significant(P < 0.05).(P < 0.05).Conclusions EGFR gene mutation rate in pleural effusion is lower than that in tumor tissue specimens in terminal NSCLC patients and tumor tissue is still the best test specimens.

Objective To explore the regulation mechanism for miR - 125a - 5P in epidermal growth factor receptor(EGFR)signaling pathway in medulloblastoma. Methods The potential targets of miR - 125a - 5P in the EGFR signaling pathway were predicted by TargetScan and Sanger software,there were 3 groups:control group,non -sense group and miR - 125a - 5P group. Their relationship,between miR - 125a - 5P and cyclin - dependent kinase in-hibitor 2B( CDKN2B),E2F transcription factor 3( E2F3),mitogen - activated protein kinase 14( MAPK14)and growth factor receptor - bound protein 10(GRB10),were tested by luciferase experiments. After miR - 125a - 5P oligo-nucleotide was transfected to D341 cells,miR - 125a - 5P level was detected by reverse transcription polymerase chain reaction. Then the thiazolyl blue tetrazolium bromide assay was used to draw the cell growth curves,and Transwell assay was used to detect cell migration ability. The expression levels of GRB10,EGFR,phosphatidylinositol 3 - kinase(PI3K) and Ras were tested by Western blot method. Results The results of luciferase experimental results showed that GRB10 was the only target gene of miR - 125a - 5P. After miR - 125a - 5P being transfected,the D341 cell prolifera-tion obviously declined markedly. Compared with control group[(38. 16 ± 7. 47)% ]and the non - sense group [(36. 79 ± 8. 94)% ],cell migration rate in the miR - 125a - 5P group was lowest[(13. 59 ± 4. 41)% ],and there was a significant difference among 3 groups(χ2 = 11. 495,P < 0. 05);in the miR - 125a - 5P group,the expression level of EGFR increased 1. 67 times,GRB10,PI3K and Ras levels were reduced to 23% ,61% and 42% . Conclusion miR - 125a - 5P can inhibit tumor growth by silenced GRB10 expression targeting EGFR downstream signaling pathways in medulloblastoma.

BACKGROUNDIn March 2013, human cases of infection with a novel A (H7N9) influenza virus emerged in China. The epidemic spread quickly and as of 6 May 2013, there were 129 confirmed cases. The purpose of this study was to analyze the epidemiology of the confirmed cases, determine the impacts of bird migration and temperature changes on the H7N9 epidemic, predict the future trends of the epidemic, explore the response patterns of the government and propose preventive suggestions.

METHODSThe geographic, temporal and population distribution of all cases reported up to 6 May 2013 were described from available records. Risk assessment standard was established by analysing the temperature and relative humidity records during the period of extensive outbreak in three epidemic regions in eastern China, including Shanghai, Zhejiang and Jiangsu provinces. Risk assessment maps were created by combining the bird migration routes in eastern China with the monthly average temperatures from May 1993 to December 2012 nationwide.

RESULTSAmong the confirmed cases, there were more men than women, and 50.4% were elderly adults (age >61 years). The major demographic groups were retirees and farmers. The temperature on the days of disease onset was concentrated in the range of 9°C-19°C; we defined 9°C-19°C as the high-risk temperature range, 0°C-9°C or 19°C-25°C as medium risk and <0°C or >25°C as low risk. The relative humidity on the days of disease onset ranged widely from 25% to 99%, but did not correlate with the incidence of infection. Based on the temperature analysis and the eastern bird migration routes, we predicted that after May, the high-risk region would move to the northeast and inland, while after September, it would move back to north China.

CONCLUSIONSTemperature and bird migration strongly influence the spread of the H7N9 virus. In order to control the H7N9 epidemic effectively, Chinese authorities should strengthen the surveillance of migrating birds, increase poultry and environmental sampling, improve live poultry selling and husbandry patterns and move from a "passive response pattern" to an "active response pattern" in focused preventive measures.

Animals ; Birds ; China ; epidemiology ; Influenza A Virus, H7N9 Subtype ; pathogenicity ; Influenza in Birds ; epidemiology ; Temperature

BACKGROUNDIn 2012, the working group on abdominal problems of the European Society of Intensive Care Medicine (ESICM) proposed a definition and also guidelines for the grading system and treatment of acute gastrointestinal injury (AGI). Until now, clinical reports on this topic have not been available, and the practicality of using the AGI grading system requires further validation in the clinic. Therefore, we conducted this study to evaluate the feasibility of utilizing the current AGI grading system in a clinical environment, and to provide evidence for its usefulness in assessing the severity and prognosis of critically ill patients with gastrointestinal dysfunction.

METHODSA total of 133 patients were examined for the presence or absence of AGI, their scores on the Acute Physiology and Chronic Health Evaluation (APACHE) II and Lausanne Intestinal Failure Estimation (LIFE) test, and 28 days mortality. The presence and severity of AGI was based on guidelines provided by the ESICM. The patients were assigned to a NO-AGI group (n = 50) or an AGI group (n = 83). The AGI group was then further divided into three subgroups, consisting of AGI I (risk group, n = 38), AGI II (gastrointestinal dysfunction group, n = 33) and AGI III+AGI IV (gastrointestinal failure group, n = 12). These subgroups were then compared for differences in AGI indicators.

RESULTSThere were no statistically significant differences between the AGI group and the NO-AGI group in terms of age, gender, APACHE II score or LIFE score (P > 0.05); however, the two groups showed a significant difference in their respective rates of 28 days mortality (32.5% in the AGI group vs. 8.0% in the NO-AGI group (P < 0.05)). Patients in the three AGI subgroups showed significant differences in their 28 d mortality rates, APACHE II, and LIFE scores. AGI grading system showed strong positive correlations with APACHE II and LIFE scores (P < 0.05).

CONCLUSIONSThe current AGI grading system can be used to identify and evaluate gastrointestinal dysfunction in critically ill patients, and also to provide a preliminary assessment regarding the prognosis for patients with different grades of AGI.

Abdominal Injuries ; diagnosis ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Critical Care ; Critical Illness ; Female ; Humans ; Intensive Care Units ; Male ; Middle Aged ; Multiple Organ Failure ; diagnosis ; Prognosis ; Thoracic Injuries ; diagnosis ; Young Adult

Objective To explore the expression of HCCR, pERK and pELK1 in gastric cancer tissue and para-carci-noma tissue, and to explove its correlations. Methods In 60 human gastric cancers and their corresponding paraneo-plastic tissuses,the expression of HCCR, pERK and pELK1 were detected by immunohistochemistry. Results The ex-pression of HCCR was 75.0%(45/60), pERK, and pELK1 were 73.3%(44/60) and 71.7%(43/60), respectively, and the expression of proteins in gastric tumor were higher than that in paraneoplastic tissues (P<0.05). Conclusion The HCCR, pERK and pELK1 are higher expressed in tumor tissues, availably as a auxiliary index for clinical immunohis-tochemistry in gastric cancer.

Objective To evaluate the clinical outcomes of limited open reduction via sinus tarsi approach and traditional open reduction internal fixation of the treatment for Sanders type Ⅱ calcaneal fracture.Methods Between February 2010 and February 2011,30 patients were enrolled into our study and were divided into minimal invasive and traditional groups randomly.Each group consisted of 15 patients.When soft tissue swelling subsided,the patients of mininal invasive group were performed a limited ORIF via a sinus tarsi incision,while those traditional groups were performed ORIF via a classical lateral extensile L-shape approach.X-rays were taken in the regular follow-up,B(o)hler and Gissane angle were measured.The final curative effect was comprehensively assessed according to visual analogue scale (VAS),the ankle and hind-foot score of American Orthopaedic Foot and Ankle Society (AOFAS) and SF-36 at the last follow-up,with the complications recorded.Results The average time of the follow-up was 16.9 nonths and 19.9months respectively in two groups.Superficial skin necrosis occurred in 2 cases in traditional group.X-ray demonstrated bone union 3 months after the operation in both groups.And no implant failure occurred.The B(o)hler angle of minimal invasive group was 13.1°±3.8° and the traditional group was 14.9°±4.3°,the Gissane angle of minimal invasive group was 28.1°±7.8° and the traditional group was 26.2°±8.2°.The average AOFAS ankle and hindfoot score of minimal invasive group at final follow-up was 91.2±15.9,and the average VAS score was 1.7±1.3,while the traditional group was 82.4±14.7 and 1.9±2.1 respectively.But SF-36 score in minimal invasive group (79.5±12.1) was higher than that in traditional group (70.2±12.4).Four patients in minimal invasive group and 15 in traditional group suffered from varying degrees of subtalar joint stiffness.Conclusion No significant difference was found between the two groups in the short-term efficacy of the treatment for Sanders type Ⅱ calcaneal fracture.However,minimal invasive technique has the advantages of lower soft tissue complication rate and lower suhtalar joint stiffness rate.

In the process of hemodialysis, the patients' blood pressure and pulse wave are likely to change considerably so that hypotension and rapid heart rate may be dangerous to the hemodialysts' life. Based on the environment of hemodialysis room, we designed the communication system using SPCE061A single-chip computer and STC89C52 single-chip computer to realize wireless communication between upper system and the two single-chip computers, and on the upper computer the pulse wave can display at real-time and save data on the data base. At the same time we designed an interface program to reduce noise by way of frequency domain analysis and wavelet denoising. It can draw the cycle pulse wave, eigenvalue-K, the maximum peak and peak frequency, it can effectively judge a racing heart in the hemodialysis and can also evaluate vascular sclerosis of hemodialysis. In short, the system is able to improve treatment security and reduce the burden of the doctors and nurses in hemodialysis rooms.
Algorithms ; Humans ; Information Systems ; Monitoring, Physiologic ; methods ; Pulse Wave Analysis ; Radial Artery ; Renal Dialysis ; adverse effects ; Software Design