Objective To observe the NTBC dependence of Fah-knockout mice and study the biological characteristics in order to use the model more effectively.Methods Examine the progressive changes in body weight, survival time, liver pathology and serological markers after the NTBC withdrawal.Results After removing of NTBC, Fah-knockout mice lost their body weight gradually, and finally died in 5 to 7 weeks, along with increased serum ALT, AST levels and deformation of the hepatocytes.Conclusions Fah-knockout mice have a strong drug dependence of NTBC and could be the ideal model to hereditary tyrosinemia type I and other liver injury.

Objective To investigate the effect of the Kolb experience learning theory applied in Surgical Nursing teaching.Methods 120 students were selected randomly and were divided into 2 groups,the observation group operated the teaching mode based on the Kolb experience learning theory,the control group operated traditional teaching.After the experiment,using the theory test,the skill test,the questionnaire survey to evaluate the teaching effectiveness.Results The two groups showed statistical difference in theory exam results,learning effect of each dimension and evaluation of teaching effect.Conclusions The teaching mode based on the Kolb experience learning theory can improve the students' interest in learning and study effect,and push forward the reform of teaching implementatiou.

Aim To construct human phage single-chain antibody library associated with esophageal cancer and to screen the specific scFv against Eca109 cells from the liberary. Methods Metastatic periesophageal lymph nodes of esophageal cancer patients were used as the B cells source, the total RNA of these B cells was extracted and prepared as the template of RT-PCR. First, we screened graticulely two pairs of primers of the heavy and light regions separately, then the V_H and V_L fragments were first amplified from the cDNA by the polymerase chain reaction (PCR). Second, the V_H-linker and V_L-linker were amplified from the V_H and V_L fragments. Last, the V_H-linker and V_L-linker were assembled into scFv gene fragments by SOE-PCR,and then Sfi I and Not I restriction site were inlet in it. ScFv gene was cloned into the pCANTAB-5E phagemid. Phagemids were introduced into E.coli TG1 by electrotransformation, followed by rescue of antibody-expressing phage using M13K07 helper-phage superinfection. Recombinant scFv phage library was constracted and PCR was used to identify the insert ratio of scFv antibodies library. Results of SfiI/Not I double digestion reaction positive insert clone were identified by 1.5% agarose gel electrophoresis. The phage library was panned with NHEEC and Eca109 cancer cells in suspension for four rounds. Strongly positive recombinant phage clones were used to infect E.coli HB2151. Expression of soluble scFv was induced by IPTG. Soluble scFv from periplasm were purified by affinity chromatography and identified by SDS-PAGE and Western blot. Cell ELISA , immunohistochemical staining and immunocytochemical staining were used to identify the activity of the soluble scFv. Results The result of agarose gel electrophoresis showed that total RNA of these B cells had two bands of 28 S and 18 S. The size of V_H fragment is about 450 bp,V_L fragment is about 350 bp and scFv is about 850 bp. The competence is 108 cfu??g-1 pUC18 DNA. Randomly digestive reac-tion showed that the positive insert ratio was 91.7% (22/24). After four rounds of panning, the fourth phage yield is 141 times as much as that of the first one. SDS-PAGE and Western blot showed that the MW of the soluble scFv was about 30 ku and the brand of 30 ku was stained. Immunohistochemical staining showed strong stainning of the tissue of esophageal cancer, but not the liver and gastric cancer tissue. Immunocytochemical staining showed significant staining of the esophageal cancer line Eca109. The result of cell ELISA assay revealed that soluble scFv had highly specific and could combined with Eca109 cells, but not with BGC-823 and NHEEC. Conclusion A human scFv phage display library associated with esophageal cancer has been constructed successfully and the specific scFv antibody against Eca109 has been identified from the liberary.

Objective To screen human anti-hypoxia-inducible factor (HIF)-1? scFv of lung adenocarcinoma from large phage antibody library and identify the positive clones. Methods Panning of large phage antibody library against lung adenocarcinoma cell line A549 and HIF-1? was conducted respectively to select specific antibodies. E. coli HB2151 was infected to induce the expression of soluble scFv. The binding activity and specificity were tested by ELISA and immunocytochemical assay. The expression and relative molecular weight of the soluble scFv was detected by SDS-PAGE and Western blot analysis. Results After panning,the target scFv was enriched,and ELISA results showed that positive reactions to HIF-1? were detected in 5 of 10 random clones with a positive ratio of 50%. Immunocytochemical analysis showed the specific affinity of the antibodies to A549 cells. The soluble human anti-HIF-1? scFv fragments of lung adenocarcinoma were expressed in E. coli HB2151 and then confirmed by SDS-PAGE. The result of Western blotting showed that the relative molecular weight of the soluble scFv was about 30?103. The binding activity and specificity were confirmed by ELISA. Conclusion Human anti-HIF-1? scFv of lung adenocarcinoma is successfully obtained with large phage antibody library technique.

Aim To screen and identify anti-CCR7 single chain fragments variable(scFv)from lager phage library and to detect the scFv efficiency.Methods The insert ratio of ScFv antibodies library was identified by PCR.The products digested by Sfi I/Not I double enzyme.Panning against breast cancer cell and CCR7 were performed four and three rounds respectively.Positive clones were transformed to E.coli HB2151, and their dissolvability was assayed.The soluble scFv was purified by affinity chromatography, and its relative molecular mass was determined by Western blot.The ELISA assay was used to identify the immunocompetence of the antibody.Immunocytochemical staining and radioimmunoimaging were employed to determine the affinities of scFv binding with CCR7 in cell line and in nude mice.Results The insert ratio of ScFv gene was 90%(18/20), enzyme digest reaction showed the aim products on 1% agarose gel.ScFvs were obviously enriched after 7 round panning.Western blot result showed soluble scFv's molecular mass was about 34 KD.ELISA analysis showed dissolved antibody had high immunocompetence to MDA-MB-435 s cells.Immunocytochemical staining and radioimmunoimaging indicated that ScFvs were bound efficiently to MDA-MB-435 s cells which expressed CCR7.Conclusions ScFvs against CCR7 are successfully acquired by screening the phage antibody library.The soluble ScFvs have high affinity and specifical binding to human breast cancer cells.

Objective To investigate the safety and effect of high intensity focused ultrasound ablation(HIFU) on residual liver cancer in patients after radiotherapy. Methods Twenty liver cancer patients with twenty five residual tumors after three-dimensional conformal radiotherapy or stereotactic radiotherapy received HIFU ablation. Liver function and periphery blood cell counts were performed before HIFU and at 1 week after HIFU in all patients. Enhanced MRI and α-fetoprotein (AFP) level were performed before HIFU and at 2 weeks after HIFU to evaluate the effect of HIFU ablation. The survival of all patients was assessed by the Kaplan-Meier method. Results The mean follow-up time was (12.6 ± 8.0)months. The median survival time and 1-year survival rate were 22 months and 87.5% respectively. No skin burns were observed in all patients. As compared with before HIFU,there were no significant differences in the levels of albumin,alanine transarninase, aspartate transarninase, total bilirubin, direct bilirubin, blood red cell counts,blood white cell counts and blood platelet counts at 1 week after HIFU (paired t test, P =0. 156,0. 356,0. 203,0.659,0. 531,0. 519,0. 310,0. 346, respectively). Significant difference in AFP level of 9 patients with AFP＞20 μg/L was observed before HIFU and 2 weeks after HIFU (paired t test, P =0.030). Among 25 residual liver tumors,sixteen with complete ablation and six with ablation volume of ≥80% were observed by enhanced MRI at 2 weeks after HIFU. Conclusions HIFU may be a safe and effective new method to ablate residual liver cancer after three-dimensional conformal radiotherapy or stereotactic radiotherapy.

Objective: To study the pharmacokinetics and brain/plasma concentration ratio of nortriptyline at multiple doses in mice which were pre-treated with physiological saline, piperine and verapamil. Methods: A total of 216 male Kun Ming mice[(25±3) g] were equally divided into 4 groups randomly. Each group was intragastrically administered physiological saline (B), piperine (170 μg/kg), piperine (5 mg/kg) and verapamil (5 mg/kg) for 8 days. On the 8th day, 1 h atfer giving the above drugs, each mice was intraperitoneally injected nortriptyline (13 mg/kg). The mice were sacriifced by picking off eyeballs at the time intervals of 5, 15, 30 min, and 1, 2, 4, 6, 8 and 12 h, andthe cerebra were collected and weighted. Nortriptyline in mouse plasma and brain was determined by HPLC-MS/MS. The pharmacokinetic properties of the plasma, brain and brain/plasma were calculated. Results: hTe AUC0-12 h of brain/plasma concentration ratio in the 170 μg/kg piperine group was significantly lower than that in the other groups (P<0.05), while the AUC0-12 h of brain/plasma concentration ratios in the 5 mg/kg piperine group and the verapamil group were not signiifcantly different from those of untreated mice. Conclusion: Piperine (170 μg/kg) may induce P-glycoprotein expression in the blood-brain barrier, while piperines at 5 mg/kg has no influence on P-glycoprotein expression in the blood-brain barrier.

This paper introduces some information of the multi-function hydraulic pressure diagnostic and therapeutic instrument, which is applied to the diagnoses and treatments of oviduct, matrix, urethra, bladder and bile duct, including its system function, operation principle, software and hardware designs. Being reliable, cheap, convenient and multipurpose, the instrument has a bright future in the market.

This paper introduces some information of the infusion tele-control system based on LonWorks, including its system function, operation principle, software and hardware designs. With high reliability, low cost and convenient operation, this system has a bright future in the market and its application.

Objective:To evaluate the effect of Jianxin-Pinglyu Pills combined with routine western medicine therapy on the patients with diabetes mellitus and ventricular arrhythmia (VA). Methods:A total of 92 patients with type 2 diabetes mellitus (T2DM) and VA who met inclusion criteria and treated in the hospital were enrolled between June 2019 and December 2020. According to random number table method, they were divided into observation group ( Jianxin-Pinglyu Pills on basis of control group) and control group (routine treatment), 46 cases in each group. Both groups were continuously treated for 2 months. Before and after treatment, TCM symptoms (chest pain and palpitation, chest distress and shortness of breath, dry mouth and thirst, fatigue) were scored. The fingertip blood was collected to detect levels of fasting blood glucose and 2 hPG. The 24 h dynamic electrocardiogram examination was conducted by electrocardiograph. The standard deviation of NN intervals (SDNN), standard deviation average of NN intervals (SDANN), the root mean square of successive R-R interval (RMSSD) and the percentage of adjacent NN intervals over 50 ms (PNN50) were statistically analyzed. The creatine kinase (CK) and creatine kinase isoenzymes (CK-MB) were detected by biochemical analyzer. The plasma brain natriuretic peptide (BNP) were detected by chemiluminescence immunoassay. The adverse reactions were recorded, and clinical curative effect was evaluated. Results:The differences in total response rate between observation group and control group were statistically significant [93.5% (43/46) vs. 78.3% (36/46); χ2=4.390, P=0.036]. After treatment, TCM symptoms (chest pain and palpitation, chest distress and shortness of breath, dry mouth and thirst, fatigue) in observation group were significantly lower than those in control group ( t=4.319, 2.714, 3.196, 4.667, P<0.01 or P<0.05), levels of fasting blood glucose and 2 hPG were significantly lower than those in control group ( t=2.025, 6.462, P<0.05 or P<0.01). After treatment, SDNN [(119.72 ± 5.86) ms vs. (115.27 ± 6.10) ms, t=3.568], SDANN [(112.80 ± 13.36) ms vs. (98.62 ± 14.51) ms, t=4.876], RMSSD [(46.87 ± 5.44) ms vs. (42.58 ± 5.73) ms, t=3.683] and PNN50 [(6.84 ± 0.66)% vs. (6.21 ± 0.64)%, t=4.648] in observation group were significantly higher than those in control group ( P<0.01), while the levels of CK [(263.82 ± 52.18) U/L vs. (306.59 ± 48.45) U/L, t=4.074] and CK-MB [(2.02 ± 0.62) g/L vs. (3.25 ± 0.49) g/L, t=10.556] were significantly lower than those in control group ( P<0.01), and level of plasma BNP [(126.99 ± 49.02) ng/L vs. (188.26 ± 54.30) ng/L, t=5.681] was significantly lower than that of control group ( P<0.01). During treatment, difference in incidence of adverse reactions between observation group and control group was not statistically significant [10.9% (5/46) vs. 4.3% (2/46); χ2 =0.619, P=0.432) ]. Conclusion:Jianxin-Pinglyu Pills combined with routine western medicine can effectively relieve clinical symptoms in patients with T2DM and VA, control hyperglycemia, improve heart rate variability, control levels of myocardial enzymes and plasma BNP, and safety.