[Abstact] Circular RNAs(circRNAs) were once mistakenly considered as the transcription splicing intermediates , affiliate by-products or splicing errors , and the understanding of circRNAs has been significantly revolutionized with the development of high-throughput sequencing technology of RNA and the improvement of the corresponding bioinformatics analysis .CircRNAs with their unique closed-loop structure and undenible important biological functions have become new favorites in both molecular biology studies and clinical trials.To our knowledge , circRNAs usually arise after transcription , stably express in a variety of biological cells and regulate or interfere genetic expression , thus affecting occurrence and progression of diseases.Recently, circRNAs′influences upon various cancers have begun to edge up and we elaborates them in detail here based on the latest researches .

Objective To investigate the inhibitory effect of maspin gene on metastasis and invasion of gastric cancer and its mechanism.Methods Maspin gene was ligated to the expression vector PCR2.1 with T4 DNA ligase after the PCR2.1 was digested by HindⅢ/XbaⅠ.The recombinant vector maspin/PCR2.1 was transfected into human gastric cancer cell lines MKN28 and SGC7901,then the mRNA and protein expression changes of maspin gene,uPA and uPAR were detected respectively by PT-PCR and Western blotting.Results After identified by digestion and sequencing,the reconstructed plasmid was confirmed to contain the correct and full nucleotide sequence of maspin gene.The expressions of maspin gene were up-regulated in MKN28 and SGC7901 cell lines,while the expressions of uPA and uPAR were down-regulated.Conclusion The expression vector maspin/PCR2.1 is constructed successfully and can be expressed in eukaryotic cells.Expressions of uPA and uPAR can be inhibited by maspin gene.

Objective To construct a recombinant eukaryotic vector expressing maspin cDNA,and to explore the effect of maspin overexprssion on the proliferation of human gastric carcinoma cell line SGC7901.Methods The fragment of maspin gene was amplified by polymerase chain reaction(PCR).An eukaryotic vector expressing maspin(maspin/PCR2.1)was constructed with PCR2.1,and transfected into SGC7901 cells.The expression of maspin at mRNA and protein level was detected by RT-PCR and Western blotting.The proliferation of SGC7901 cells was observed by MTT.Cell cycle was analyzed by flow cytometry.Results Recombinant plasmid maspin/PCR2.1 was constructed and transfected into SGC7901 successfully.The mRNA and protein levels of maspin were significantly higher in the maspin/PCR2.1 group(33.6?1.2,23.4?1.6)than that in the blank PCR2.1(15.0?1.5,12.3?1.5)and the untreated group(13.7?2.0,12.0?1.3)(P

Objective To develop an HPLC-MS/MS method for simultaneous determination of xylazine and 2,6-xylidine in body fluid samples.Methods The samples were extracted by HLB SPE,separated by Waters Atlantis dC18 chromatographiccolumn,and then detected in the MRM scan ion mode under positive ionization condition.Results The recoveryrates of this method were between 70.5％ and 79.8％ with the range of RSD rfrom8.2％ to 10.5％.The limits of detection ofxylazine and 2,6-xylidine in blood and urine samples were 0.4 and 0.3 ng/mL,and the limits of quantitation were 1.2 and 1.0 ng/mL,respectively.Conclusion This method is of high sensitivity,good specificity and good reproducibility,and thus could besuitable for accurate quantification of xylazine in blood and urine samples.

Objective To investigate the effect of calcipotriol on melanin synthesis by human melanocytes and its possible action mechanism.Methods Primary melanocytes were cultured with various concentrations(10~(-5),10~(-6),10~(-7),10~(-8),10~(-9),10~(-10) mol/L)of calcipotriol for 24 or 48 hours.Subsequently,MTT assay,NaoH assay.Dopa-oxidase assay,Western blot and semiquantitative RT-PCR were used to measure the cell proliferation of,melanin synthesis by.tyrosinase activity,protein and mRNA expression levels in the melanocytes.respectively.Those untreated melanocytes served as the control.Results The calcipotriol between 10~(-9) and 10~(-5) mol/L had no significant effect on the proliferation of cultured melanocytes(P＞0.05).while that of 10~(-9) and 10~(-8) mol/L increased tyrosinase activity by 137%and 123%,and enhanced melanin synthesis by 40.63%and 18.75%,respectively,compamd with untreated melanocytes(both P＜0.05).Moreover,the tyrosinase protein level increased by 270.4%(P＜0.05)in melanocytes treated with calcipotriol at 10~(-9) mol/L for 24 hours.The strongest tyrosinase activity and melanin synthesis was observed in melanocytes treated with calcipotriol of 10~(-9) moI/L.Conclusions The proliferation of melanocytes is unaffected by calcipotriol at 10~(-9) to 10~(-5) mol/L,but it can elevate the expression of tyrosinase protein,enhance tyrosinase activity,and promote melanin synthesis in melanocytes.

Objective To detect the change of brain activity under different depth of anesthesia (DOA)noninvasively. Methods The Lempel-Ziv complexity C(n) was used to analyze EEG and its four components (delta,theta, alpha, beta), which was recorded from SD rats under different DOA. The relationship between C(n) and DOA was studied. Results The C(n) of EEG will decrease while the depth of anesthesia increasing and vice versa. It can be used to detect the change of DOA sensitively. Compared with power spectrum, the change of C(n) is opposite to that of power spectru,. Only the C(n) of delta rhythm has obvious variations induced by the change of DOA, and the variations of delta is as similar as the EEG's. Conclusion The study shows that the desynchronized EEG is replaced by the synchronized EEG when rat goes into anesthesia state from awake, that is just the reason why complexity and power spectrum appear corresponding changes under different DOA. C(n) of delta rhythm dynamic change leads to the change of EEG, and the delta rhythm is the dominant rhythm during anesthesia for rats.

Objective To evaluate the efficacy and safety of Sheng-qi-zhuang-yang formula combined with specific immunotherapy (SIT) with standardized house dust mite vaccine on allergic asthmatic children. Methods Participants were 100 children with mild to moderate allergic asthma , who were receiving SIT at Guangdong Provincial Hospital of Chinese Medicine from Jan 2011 to Dec 2013 , who were divided into treatment group and control group. Patients in the treatment group received Sheng-qi-zhuang-yang Decoction combined with SIT while patients in the control group received SIT alone. Asthma symptom scores, respiratory function and related adverse events were compared before and after treatment, between groups. Results The desensitization treatment functions was ahead of time than expected in both groups. There is no significant difference between groups in terms of respiratory function and adverse effects. Conclusion Sheng-qi-zhuang-yang decoction combined with SIT for allergic asthmatic children seems to advance clinical effect without increasing adverse events. Further large scale clinical trial is required to confirm this.

AIM:To investigate the role of damaged mitochondria in dendritic cell ( DC) apoptosis induced by Vibrio vulnificus (Vv) and its possible mechanism.METHODS: DC2.4 cells were co-cultured with Vv 1.1758 strain. Fluorescent probes DCFH-DA and Fluo-8-AM were used to detect reactive oxygen species ( ROS) and intracellular Ca 2+concentration in the invaded cells , respectively .The cellular apoptotic rates and mitochondrial membrane potential (Δψm ) were measured by flow cytometry.The expression of nuclear factor-kappa B p65 (NF-κB p65) and tumor necrosis factor-al-pha (TNF-α) was detected by Western blotting.RESULTS:Vv 1.1758 induced DC2.4 cell apoptosis.Vv 1.1758 bacte-ria invaded into the DC2.4 cells by binding with cellular membrane though the end of the body .In the invaded DC2.4 cells, the visible mitochondrial damage, elevated ROS and intracellular Ca2+levels, and declinedΔψm were presented.Af-ter 1 h of co-culture, NF-κB p65 began to rise and reached the peak at 5 h, and then slightly decreased at 6 h.The TNF-αlevel increased after 2 h of co-culture and reached the peak at 6 h.CONCLUSION:The damaged mitochondria play an important role in DC apoptosis induced by Vv , and its possible mechanism may associate with the elevation of ROS and in-tracellular Ca2+level, and the declined Δψm.Meanwhile, NF-κB p65 and TNF-αare potential critical signaling molecules in the process of apoptosis .

Objective To evaluate the effects of different target effect-site concentrations (Ces) of sufentanil on the minimum alveolar concentration of desflurane inhibiting stress responses to skin incision (MACsAR) in 50％ of patients undergoing abdominal surgery.Methods Eighty-three patients,aged 20-60 yr,scheduled for elective abdominal surgery with an expected incision longer than 10 cm,were enrolled in the study.All the patients were randomly allocated to one of three groups using a random number table:control group (group C,n =28),sufentanil with target Ce of 0.1 ng/ml group (group S1,n =27),and sufentanil with target Ce of 0.3 ng/ml group (group S2,n =28).After tracheal intubation,desflurane inhalation was started,and sufentanil was infused at the preset target Ces in S1 and S2 groups.The initial end-tidal concentration of desflurane was 9.0％ in group C,and 6.0％ in S1 and S2 groups.The target concentration was maintained at least for 15 min.The modified up-and-down method was used to perform the test.The end-tidal concentration of desflurane was adjusted in the next patients according to the response to skin incision.The MACBAR and 95％ confidence interval of desflurane was calculated according to the up-and-down method.Mean arterial pressure and heart rate were monitored and recorded and rate-pressure product was calculated.Results The MACBAR (95％ confidence interval) ofdesflurane was 11.2％ (11.1％-11.3％),7.8％ (7.7％-7.9％),and 4.2％ (4.1％-4.3％) in C,S1 and S2 groups,respectively.MACBAR and rate-pressure product of desflurane were significantly lower in S1 and S2 groups than in C group,and in S2 group than in S1 group (P ＜ 0.05 or 0.01).Conclusion The optimum target Ce of sufentanil is 0.3 ng/ml when combined with desflurane in the patients undergoing abdominal surgery.

Objective To clone, express and purify the extracellular carbohydrate recognition do-main(CRD) of Dectin-1 in mouse peritoneal macrophages and to further investigate its ability to recognize and bind to β-glucans. Methods The Dectin-1 CRD gone was amplified by RT-PCR from RNA of mouse peritoneal macrophages and cloned into prokaryotic expression vector pET28a (+), the constructed pET-CRD recombinant plasmid was transformed into Escherichia coli BL21 (DE3) and the fusion protein was in-duced to express. After affinity purification and renaturation, the fusion protein was incubated with Candida albicans yeast and its ability to recognize and bind to β-glucans in the cell wall of fungi. Results The fu-sion protein could recognize β-glucans in the fungal cell wall. Conclusion The recombinant expression plasmid pET28a-CRD was successfully constructed and the fusion protein was induced. The fusion protein is able to recognize and bind to β-glucans in the fungal cell wall, thus laying a good foundation for fungal de-tection and the exploration of the biological role of β-glucans.