Teaching evaluation from the students is the necessary key point in the management process of teaching quality in colleges. This essay introduces the design and development of the system of teaching evaluation on net in colleges about the technology of data base of ASP NET and JAVA. This system will overcome the deficiency of the traditional evaluation model in teaching,which remains the short comings of too large a number of data,long time in data processing,as well as the low working efficiency. It also probes on the application of the students’ evaluation system.

microRNAs is a group of small non-coding RNAs that play a negative regulation role in expression of target genes at post-transcriptional level by inhibition or degradation of target mRNAs after combination of the seed sequence (SS) in microRNAs with the SS-binding sequences usually located at 5'ends of target mRNAs.microRNAs was firstly found in Caenorhabditis elegans.Subsequently,many different microRNAs in eukaryocytes were revealed.In eukaryocytes,microRNA precursors are transcribed at first and then become functional microRNAs with 21-23 nt in size after splice.Most of eukaryocytic microRNAs combime with the sequences at 3'end of target mRNAs that cause the translation inhibition or degradation of the mRNAs.In the recent years,many different prokaryocytes,such as bacteria,have been confirmed to possess microRNAs.However,the microRNAs in prokaryotes such as bacteria are 50-400 nt in size and have the biological activity without splice.Moreover,the characteristics,action sites and mechanisms of the prokaryotic microRNAs have some certain diversity compared to the eukaryotic microRNAs.Our review briefly introduce the major regulation mechanisms of gene expression as well as the general characteristics of microRNAs and their regulation mechanisms of gene expression in prokaryocytes and eukaryocytes,which will provide a basis for further and profound study on the gene expression regulation and pathogenic mechanisms of prokaryotic microbial pathogens.

Objective To investigate the correlation between Streptococcus pneumoniae (S.pneumoniae) StkP kinase and drug resistance and to analyze the binding ability of StkP extracellular region (EC-StkP) to β-lactam antibiotics.Methods A stkP gene knockout (ΔstkP) mutant was constructed from S.pneumoniae strain ATCC6306 by insertional inactivation method.E-test was performed to detect the minimum inhibitory concentrations (MIC) of penicillin (PCN) and cefotaxime (CTX) against ΔstkP mutant and its wild-type strain.Bioinformatic softwares were used to predict the EC-StkP of S.pneumonia strain ATCC6306,to generate the three-dimensional structure model of EC-StkP and to analyze the correlation between the structure and functions of EC-StkP.PCR was performed to amplify the extracellular segment of stkP (EC-stkP) gene and the product of it was sequenced after T-A cloning.A prokaryotic expression system of EC-stkP gene was constructed.SDS-PAGE in combination with a gel image analysis system was used to detect the expression of the recombinant EC-StkP (EC-rStkP).The expressed EC-rStkP was extracted by Ni-NTA affinity chromatography.The binding abilities of EC-rStkP to PCN and CTX were detected by isothermal titration calorimetry (VT-ITC) and surface plasmon resonance (Biacore).Results S.pneumonia strain ATCC6306 was sensitive to PCN (MIC=0.06 μg/ml) and CTX (MIC=0.12 μg/ml),but its ΔstkP mutant was resistant to the two antibiotics (PCN MIC=16 μg/ml,CTX MIC=32 μg/ml).The 295 aa segment was predicted as the extracellular region at C-end of StkP of S.pneumoniae strain ATCC6306,containing four penicillin-binding proteins and Ser/Thr kinase-associated (PASTA) domains.The cloned EC-stkP segment and the EC-stkP segment in GenBank shared 99.6% similarity in nucleotide sequence and 100% in amino acid sequence.The constructed prokaryotic expression system for EC-stkP gene expressed EC-rStkP in soluble form.Both PCN and CTX could bind to EC-rStkP and CTX was better than PCN in term of binding ability.Conclusion The stkP gene of S.pneumonia is closely related to drug resistance and the encoded protein,Ser/Thr kinase StkP,can recognize and bind to β-lactam antibiotics.

Background and purpose:Inhibitor of apoptosis-stimulating protein of p53 (iASPP) is one of the ASPP family. It binds to p53 to inhibit the transcriptional activity of p53-target genes and cell apoptosis, which is asso-ciated with tumor formation. Previously, we found a new subtype of iASPP, iASPP splice variant (iASPP-SV), which is a nuclear protein containing 407 amino acid residues and can bind to p53, inhibiting p53 transcriptional activity. However, the relationship of iASPP-SV and breast cancer is still obscure. Therefore, the purpose of this research was to study the role of iASPP-SV on breast cancer tumorigenesis and progression.Methods:5’-rapid ampliifcation of cDNA ends (RACE) was used to identify the 5’-end of iASPP-SV mRNA in MCF-7 cells. HEK 293 cells were transfected with pFLAG-iASPP-SV and pFLAG-iASPP (828). Then Western blot was used to identify whether endogenous iASPP-SV was expressed in HEK 293 cells and 8 types of human tumor cell lines. This study established the stable clones of NIH 3T3 expressing FLAG-iASPP-SV and FLAG-iASPP (828). Cell proliferation assay, colony formation and soft agar colony formation assay were used to identify whether iASPP-SV and iASPP (828) can promote cell proliferation and iASPP-SV is an oncogene. Real-time lfuorescent quantitative polymerase chain reactive (RTFQ-PCR) was used to de-tect the levels of iASPP-SV and iASPP (828) mRNA in primary breast cancers. Luciferase assays were used to identify the relationships between iASPP-SV, iASPP (828), p53 and NF-κB p65.Results:The study identiifed that iASPP-SV was encoded by previously reported NF-κB p65 subunit (RelA)-associated inhibitor (RAI), and endogenously expressed in many human cancer cell lines. Analysis of cell proliferation, colony formation assay and soft agar assay for colony formation identiifed that similarly to iASPP (828), iASPP-SV promoted tumor cell proliferation and acted as an onco-gene. RTFQ-PCR result showed that the median values of iASPP-SV and iASPP (828) in breast cancers with wild-type p53 were more signiifcantly over-expressed than those of mutant p53. Luciferase assays showed that iASPP-SV and iASPP (828) could suppress NF-κB p65 transcriptional activity. Thus iASPP family may participate in the regulation of p53 and NF-κB activity, which imply that iASPP perhaps shows pro- or anti-survival activities when it interacts with different proteins.Conclusion:These ifndings indicate that iASPP-SV may be a potential target for breast cancer thera-py.

Objective:To compare the efficacy of different concentrations of ropivacaine for interscalene brachial plexus block in patients undergoing arthroscopic shoulder surgery under general anesthesia.Methods:Ninety American Society of Anesthesiologists physical statusⅠor Ⅱ patients (NYHA classⅠorⅡ) of both sexes, aged 18-64 yr, with body mass index of 18.0-26.9 kg/m 2, undergoing elective arthroscopic shoulder surgery were selected, and were divided into 3 groups ( n=30 each) using a random number table method: 0.25% ropivacaine group (group A), 0.375% ropivacaine group (group B) and 0.5% ropivacaine group (group C). Interscalene brachial plexus block was performed with 0.25%, 0.375% and 0.5% ropivacaine 20 ml in A, B and C groups, respectively.Before operation (T 0) and at 30 min (T 1), 4 h (T 2), 6 h (T 3), 8 h (T 4), 10 h (T 5) and 12 h (T 6) after administration, the diaphragmatic mobility was measured and recorded using M-mode ultrasound and forced expiratory volume in the first second (FEV 1) and forced vital capacity (FVC) were measured using portable spirometer.The occurrence of phrenic paralysis was recorded at T 1-6.The duration of sensory and motor block was recorded.When visual analogue scale score>3 within 24 h after operation, flurbiprofen axetil 50 mg was injected intravenously for analgesia and the consumption was recorded.The adverse reactions such as cardiovascular events, local anesthetic intoxication, Horner syndrome, pneumothorax, and nausea and vomiting within 24 h after administration were recorded. Results:Compared with group A, the diaphragmatic mobility was significantly decreased during quiet breathing at T 1-3 and was decreased during deep breathing at T 2-5, and the diaphragmatic paralysis rate was increased during quiet and deep breathing at T 2-3 in group B, diaphragmatic mobility was decreased during quiet and deep breathing at T 1-6, diaphragmatic paralysis rate was increased during quiet and deep breathing at T 1-4, FEV 1% and FVC% were decreased at T 1 and FVC% was decreased at T 2 in group C, and the duration of sensory and motor block was prolonged in B and C groups ( P<0.05 or 0.01). Compared with group B, the diaphragmatic mobility was significantly decreased during quiet breathing at T 4-6 and was decreased during deep breathing at T 1-6, the diaphragmatic paralysis rate during quiet breathing was increased at T 2-4 ( P<0.05) was increased during deep breathing at T 3-4, and FEV 1 % and FVC % at T 1 were decreased in group C ( P<0.05). There was no significant difference in the postoperative requirement for flurbiprofen axetil and the incidence of adverse reactions within 24 h after administration among the 3 groups ( P>0.05). Conclusion:0.25% ropivacaine 20ml provides better efficacy when used for interscalene brachial plexus block in the patients undergoing arthroscopic shoulder surgery.

Objective To establish osteochondral scaffolds with remained calcified cartilage zone for finding an ideal scaffold for tissue engineered repair of osteochondral defect.Methods Cartilage zone was harvested from fresh adult porcine knee to fabricate type Ⅱ collagen hydrogel.Bone blocks measuring 8 mm in diameter with calcified cartilage zone were prepared by trephine and acellular treatment was performed.Histological staining was used to identify complete removal of cells.To fabricate the osteochondral models containing calcified cartilage zone,type Ⅱ collagen was seeded onto the acellular bone blocks with calcified cartilage zone,lyophilized,and cross-linked with 10 g/L of genipin ethanol solution.Then cell seeding and scanning electron microscope were performed after the models were established.Results HE staining,toluidine blue staining,solid green staining,safranin O staining,and DAPI staining showed cells were completely removed from bone blocks by decellularized process.Porosity of type Ⅱ collagen sponge was (91.1 ±3.8) ％ and pore size was (79.7 ± 17.1) μm.Porosity of acellular bone blocks was (73.5 ±2.6)％ and pore size was (470.2 ± 158.8) μm.Cells seeded onto osteochondral scaffolds grew well by scanning electron microscope.Conclusion Osteochondral scaffolds with calcified cartilage zone provide good biocompatibility and suitable pore size and porosity and may be an ideal material for repairing osteochondral defect in tissue engineering.

Objective To compare the effects of thoracoscopic 2 hole and 3 hole for congenital pulmonary bulla resection.Methods 38 cases of congenital pulmonary bulla patients,both in the VATS downlink congenital pulmonary bulla resection.According to the number of holes,thoracoscopic operation were divided into the two groups. To observe the use of group 19 cases of 2 holes,19 cases 3 holes were adopted in the control group.Pull the chest tube operation time,operation time of the two groups were compared after.and the average hospitalization time after opera-tion,postoperative analgesia drug application.Results The observation group operation time,operation time,pulling the chest tube after operation the average hospitalization time, analgesic drug application rate respectively were (46.89 ±9.11)min,(3.95 ±0.85) d,(7.37 ±1.34) d,21.1%,The control group were (66.05 ±12.09) min, (4.37 ±0.98)d,(7.32 ±1.57)d,52.6%.There were statistically significant differences in rate of the two groups in operation time,analgesic drug application.(t=-5.516,χ2 =4.071,P<0.05);38 cases were cured,followed up for 3-24 months,no recurrence occurred in 1 cases.Conclusion Video assisted thoracic descending congenital pulmona-ry bulla resection,the 2 hole 3 hole more than minimally invasive,short operation time,postoperative analgesic use rate is low.

Objective To construct a ciaR gene-knockout (ΔciaR) mutant of Streptococcus pneu-moniae ( S. pneumoniae) and to investigate the effects of CiaR in CiaH/CiaR, a streptococcal two-component signal-transducing system, on the expression of genes encoding penicillin-binding proteins ( pbps genes) and cia-dependent small RNAs (csRNAs). Methods Electrophoretic mobility shift assay (ESMA) was per-formed to detect the recombinant CiaR (rCiaR)-binding pbps genes. A suicide plasmid pEVP3ciaR for ciaR gene knockout was constructed and then aΔciaR mutant was obtained through homologous recombination and insertion inactivation of the suicide plasmid, and screening with chloromycin. The mutant was identified using PCR and sequencing analysis. E-test was used to detect the minimal inhibitory concentrations ( MIC) of penicillin ( PCN) and cefotaxime ( CTX) against S. pneumoniae strains. Changes and differences in the expression of pbps genes and csRNAs in theΔciaR mutant and its wild-type strain before and after treatment with 1/4 MIC of PCN or CTX were detected using real-time quantitative RT-PCR. Results The rCiaR could bind to the promoter regions in pbp1a, pbp1b and pbp2b genes of S. pneumoniae. The ciaR gene in ΔciaR mutant was inactivated by insertion according to the results of PCR and sequencing analysis. After treatment with 1/4 MIC of PCN or CTX, the expression of pbps genes at mRNA level ( pbps-mRNAs) in theΔciaR mu-tant was significantly increased (P<0. 05), but the levels of csRNAs were significantly decreased (P<0. 05);whereas a significantly decreased pbps-mRNAs (P<0. 05) and increased csRNAs (P<0. 05) were observed in its wild-type strain. The result of E-test showed that the MICs of PCN and CTX against ΔciaR mutant were increased by 250-fold as compared with those against its wild-type strain. Conclusion The CiaR can enhance the drug resistance of S. pneumoniae to PCN and CTX through down-regulating the expres-sion of PBP1a, PBP1b and PBP2b and up-regulating the expression of csRNAs to inhibit the expression of PBPs.

Objective To investigate the clinical feature of perioperative management of non-small cell lung cancer(NSCLC) in aged patients and improve the efficacy of surgical treatment.Methods The clinical data of 35 aged patients with NSCLC were retrospectively analyzed.The risk factors of postoperative complications were analyzed by single factor analysis,the factors had statistical significance were included in Logistic regression analysis.Results Postoperative complications occurred in 10 cases,accounting for 28.6％,and 1 case died,accounting for 2.9％.Logistic regression analysis showed that smoking,chronic bronchitis,coronary heart disease,pulmonary lobectomy were independent risk factors of postoperative complications.ConCLusion Correct staging before operation,strict surgical indication,choose the standard surgical method to reduce surgical trauma,strengthen the perioperative management can still achieve satisfactory therapeutic effect in surgical operation for aged patients with NSCLC.

Objective:Semaphorin 4D (Sema4D) acts as a regulator for axon guidance in central nervous system development. However, new evidence indicates that Sema4D has a previously unrecognized function, namely, compensatory angiogenic factor. This study aimed to investigate the effect of Sema4D on tumor growth and vascularity of colorectal carcinoma (CRC) in nude mice. Meth-ods:Sema4D was knocked down in CRC cells by infecting the cells with lentiviruses coding for Sema4D shRNA. Two groups of cells, namely, those infected with control viruses and those infected with Sema4D shRNA viruses, were subjected to migration assay to test their ability induce endothelial cell migration. The two cell groups were subcutaneously injected into nude mice. Tumor growth was documented, and the tumors harvested from the mice were subjected to immunohistochemistry or immuno fl uorescence analyses. Re-sults:In vitro migration assay results indicated that media conditioned by HCT-116 cells infected with Sema4D shRNA lentiviruses in-duced low endothelial cell migration. The two groups of subcutaneously inoculated cells showed 100%tumorigenicity. However, tumor growth rates were significantly different between the two groups. Xenografts in which Sema4D was downregulated showed marked re-duction in tumor size and vascularity. Conclusion:Cancer cells may highly express Sema4D to trigger net neo-angiogenesis and gener-ate a tumor blood supply system. Thus, Sema4D could potentially be a target in anti-angiogenic therapy of CRC patients.