The directed molecular evolution of enzyme in vitro can not only improve the efficiency of evolution, but also evolve enzyme according to the investigator's desire. This review summed up the feasible methods of this novel technique.

Objective To evaluate the serum Krebs von den Lungen (KL)-6 for the diagnosis of interstitial lung disease (ILD) associated with connective tissue diseases (CTD) and its lung-CT subtypes.Methods Seventy-five CTD patients were employed for this study,44 CTD with ILD and 31 ILD without ILD.The 44 CTD patients with ILD were further divided into different subgroups based on lung-CT imaging and clinical indexes.The enzyme-linked immune sorbent assay (ELISA) was used to measure the serum KL-6 level.For those data that was abnormally distributed,the differences between groups was compared with independent samples nonparametric tests.Results The level of serum KL-6 in the CTD with ILD was significantly higher than that without ILD [(1 118±877) U/ml vs (253±144) U/ml] (Z=-6.047,P＜0.01).By using a criterion of 500 U/ml,our data suggested that the serum KL-6 level was useful for the ILD-CTD diagnosis;the sensitivity,specificity,positive and negative predictive values were 72.7％,87.1％,88.9％ and 69.2％,respectively.The serum KL-6 level,however,showed no statistical differences between ILD subtypes,i.e.,usual interstitial pneumonia (UIP),nonspecific interstitial pneumonia (NSIP) and indeterminate [(1 104±843) U/ml,(1 242±1 039) U/ml,(815±400) U/ml,respectively] (x2=0.35,P=0.84).Our data further showed that the KL-6 level was significantly higher in CTD-ILD patients with intensive lung lesions than those with limited lung lesions [(1 910±918) U/ml vs (459±268) U/ml] (Z=-4.364,P＜0.01).In addition,the KL-6 level was significantly higher in active ILDs than in inactive ILDs[(1 478±917) U/ml vs (598±475) U/ml] (Z=-3.915,P＜0.01).Conclusion The serum KL-6 is a valuable biomarker for CTD-ILD diagnosis and even for the assessment of the extent and activity of lung damage.

Objective To study the changes of learning-memory ability and synapse of hippocampus after radiation injuries. Methods Sprague-Dawley rats were grouped according to radiation dose to 20Gy group,30Gy group radiated by linear accelerator and control group were used before radiation and 120 days after radiation. Morris maze test were taken to study the learning and memory ability of rats in each group. Average escape latency and search strategy were scaled and analyzed in each group. The parameters of synapse in CA3 area of hippocampus were studied by using electron microscope and image analyzer. Results AEL of 20Gy group was (41. 17 ±10.76 ) s and score of SS was 27.13 ± 2.34 after 120 days' radiation but AEL of 30 Gy group was (78.49 ± 9.32)s and the score of SS was (23.19 ± 7.65 ) nm. There were significant statistic differences Compared with control group and before radiation (P ＜ 0.05 ). The thickness of PSD of 20 Gy group was ( 22.03 ± 6.84 ) nm after 120days' radiation and (23.19 ± 7.65 )nm in 30 Gy group. There were significant statistic differences compared with control group and before radiation. It was observed that both in 20 Gy and 30 Gy group' s the length of synaptic activity area was shorter,the curvature of synaptic interface was smaller,the width of synaptic cleft and the thickness of PSD was narrower than that of control group. Conclusion There was close relation between the changes of learning-memory ability and synapse of hippocampus after radiation injuries.

Objective To compare the clinical therapeutic effects of continuous veno-venous hemofiltration (CVVH) and continuous veno-venous hemodiafiltration (CVVHDF) for treatment of patients with multiple organ dysfunction syndrome (MODS).Methods A prospective observation was conducted, seventy patients with MODS admitted to the Department of Critical Care Medicine of the Affiliated Hospital of Zunyi Medical College from September 2013 to December 2016 were enrolled, and they were divided into a CVVH group and a CVVHDF group according to different treatment, 35 cases in each group. In the CVVH group, the ultrafiltration fluid flow rate was set at 1800 mL/h, while in the CVVHDF group, the flow rate was set at 2000 mL/h for both substitution fluid and dialysate, and the blood flow of the two groups was 150-180 mL/min. The changes of creatinine (SCr), urea nitrogen (BUN), pH value, HCO3-, helper T cell (Th1, Th2) and Th1/Th2 ratio were compared between the two groups before and after treatment for 24, 48 and 72 hours.Results With the extension of time, SCr, BUN were reduced in both groups after treatment compared to those before treatment, 72 hoursafter treatment they reached the lowest value, and the degree of change in CVVHDF group was more significant than that in CVVH group [SCr (μmol/L): 150.62±32.09 vs. 180.41±30.52, BUN (mmol/L): 7.31±2.19 vs. 9.06±2.36. allP < 0.05], after treatment; the pH value, HCO3- level in the two groups had a tendency at first elevated and then lowered, 48 hours after treatment they reached the peak values, compared with those before treatment, the differences were statistically significant [CVVH group: the pH value 7.42±0.08 vs. 7.25±0.10, HCO3- (mmol/L) was 22.49±5.11 vs. 15.65±4.16; CVVHDF group: pH value 7.41±0.04 vs. 7.24±0.11, HCO3- (mmol/L) 23.24±4.78 vs. 15.65±4.16, allP < 0.05], 72 hours after treatment, they began reduced (CVVH group: pH value, HCO3- were 7.39±0.09, 22.35±4.91 respectively, CVVHDF group: pH value, HCO3- were 7.38±0.06, 23.13±4.61 respectively), but they were still significantly higher than those before treatment, and at each time point compared between the two groups, no statistical significant difference was seen (allP > 0.05). With the extension of therapeutic time, Th1, Th1/Th2 in two groups were gradually elevated after treatment, and 72 hours after treatment they reached the peak values; Th2 showed a trend of gradual decrease and after 72 hours of treatment it reached the lowest value, and the changes in CVVHDF group were more significant than those in CVVH group [Th1 (×10-2): 1.51±0.12 vs. 1.21±0.11, Th2 (×10-2): 1.64±0.65 vs. 1.70±0.18, Th1/Th2: 0.91±0.14 vs. 0.71±0.15, allP < 0.05].Conclusion Both CVVH and CVVHDF can effectively remove inflammatory mediators and metabolic products, adjust the balance of acid and base, regulate immune system in the body of patients with MODS, and the CVVHDF is more effective than CVVH.

Objective To study the clinical effect of personalized intervention on elderly patients with colostomy after Miles operation for rectal carcinoma. Methods 114 cases elderly patients with colonic stoma after Miles operation for rectal cancer from June 2014 to January 2016 were divided into control group and observation group by random number method, 57 cases in each. The control group were treated with routine intervention, while patients in observation group was treated with personalized intervention. the self-care ability score before intervention, the incidence of complications, intervention satisfaction and life quality score were compared between the two groups at the same time. Results The total complication rate in the observation group was significantly lower than that in control group ( 5.25% vs 22.80%) (χ2 = 8.36, P = 0.000); the intervention satisfaction of the observation group was significantly higher than that in control group (91.23% vs 75.44%) (χ2 = 6.60, P = 0.010). After intervention, the self-care ability score and life quality score of observation group were significantly higher than that in control group (P < 0.01). Conclusion The personalized intervention can reduce the complications, significantly improve the patients' life quality score and self-care ability, and effectively alleviate the negative situation, improve the patients intervention satisfaction, with a higher development value, it is worth of clinical promoting.

Objective To establish an ELISA for detecting antibody against Aspergillus fumigatus pectate lyase A(anti-PlyA),and evaluate its diagnostic value for invasive aspergillosis (IA).Methods An indirect ELISA for IgG antibody a-gainst PlyA was established using PlyA as coating antigen.The serum from 97 IA patients,80 non-IA patients and 200 healthy donors were tested,the results were compared with anti-DPPV (antibody against Aspergillus fumigatus dipeptidyl peptidase V fragment)and anti-TR (antibody against Aspergillus fumigatus thioredoxin reductase).Results The intra-as-say coefficients of variation of the ELISA method for detecting anti-PlyA was 5.3%,and inter-assay coefficients of variation was 10.9%.The sensitivity and specificity of diagnosis of IA were 62.9% and 90.4%,respectively.The positive rates of an-ti-PlyA in non-neutropenic and neutropenic IA patients were 43.8% and 72.3%,respectively (χ2 =7.493,P <0.05).There was no significant difference between the positive rates of anti-PlyA (62.9%),anti-TR (60.8%),and anti-DPPV (67.0%) (χ2 =0.562,P > 0.05).When combined anti-PlyA,anti-TR,and anti-DPPV,the diagnostic sensitivity for IA patients in-creased to 92.8%.Conclusion An ELISA for detecting anti-PlyA was successfully established.The diagnostic value of these three kinds of antibody was superior in non-neutropenic IA patientsto that in neutropenic IA patients.The combined detec-tion of three antibodies could provide higher sensitivity.

Objective To explore the analgesic effect of ropivacaine combined with sufentanil in epidural anesthesia on walk on painless labor.Methods 134 cases of maternal pregnancy were selected and randomly divided into two groups.61 cases were treated with conventional delivery,73 cases in the experiment group were treated with walk on painless labor.The visual analogue scale and improved Bromage classification score were compared at the first stage of labor,the second stage of labor,the third stage of labor.Results Compared with the control group,the rate of cesarean section was higher(P<0.05),The visual analogue scale was higher at the first stage of labor,the second stage of labor,the third stage of labor,the duration of labor were lower at the first stage of labor,the second stage of labor,the third stage of labor(P<0.05),and there was no significant difference between the two groups in the motor nerve block.Conclusion Ambulatory labor analgesia on maternal effect significantly,can significantly reduce the pain degree of maternal birth process and deliverry time,postpartum less complications.

Objective:To study the clinical features and treatment outcome of AIDS associated oral candidiasis.Methods:The clinical data of 31 cases with AIDSassociated oral candidiasis from 201209 to 201303 were studied retrospectively,including general data,clinical features,oral manifestation,CD4 cell count,opportunistic infections,and antifungal therapy outcome,etc.Results:CD4cell count ＜200 cell/μl was found in 30 cases,AIDSrelated multiple opportunistic infection was observed in 29 cases.30 cases hadpseudomembranous candidiasis,1 cases had erythematous candidiasis and 2 cases had pseudomembranous candidiasis with angular candidiasis.After antifungal treatment,the lesion of 8 cases reduced,that of 23 cases disappeared completely,lesion relapse after drugwithdrawal happened in 3 cases.Conclusion:AIDSassociated oral candidiasis was more common in AIDS patients with CD4 ＜200cells/μl,the main clinical form is pseudomembranous type,and with multiple opportunistic infections.The antifungal treatment is effective for the patients.

Objective To investigate the effect of meisoindigo inducing primary K562 cell apoptosis and its influence on the expression of conduction signal STAT5.Methods K562 cell was treated with meisoindigo for 12 h、24 h、48 h respectively.Flow cytometry was used to detect cell apoptosis and cell cycle,and Western blot was employed to examine the alteration of the expression of STAT5.Results ①The rate of K562 cell apoptosis treated with meisoindigo for 12 h、24 h、and 48 h was 12.5 ± 0.69,19.4± 1.07 and 42.5 ± 3.22,showing statistical significance compared with the control group (P＜0.05).②Meisoindigo could block K562 cell at the S stage,the ratio of S stage cell was increased to 70.48±6.34 with the decrease of cells at G0/G1 and G2/M stage.③Gradually reduced expression of STAT5 showed after K562 cells treated with meisoindigo from 12h to 48h.Conclusion Meisoindigo accelerrates the apoptosis of K562 cell and the mechanism may relevant to the decreased expression of STAT5.

Objective To describe the understanding level of disease of children with leukemia and their ways of obtaining disease information in order to help nurses and parents to select appropriate content and manners to communicate with children about disease information.Methods In-depth interviews were conducted with 25 children' parents using a descriptive qualitative research method,and the data were analyzed using content analysis.Results Children during remission stage of leukemia had different understanding levels of their disease.Ways of children with leukemia to obtain disease information was correlated with their mental maturity.Conclusions Disease information should be told according to children's age,disease course and level of thinking,and health professionals and parents could provide appropilate ways of obtaining information for children on basis of their mental maturity.