Objective To understand the training status and learning requirement of nursing personnel for the aged from the perspective of nursing targets and their relatives in the city of Nanjing.Methods The questionnaire involved six categories,80 questionnaires were sent out,57 of them were recollected.The effective recovery rate was 71.3％(57/80).All the data were input into Excel to do statistical analysis.Results The nursing personnel did not fully grasp the skills of transfusion nursing and muscle injection [45.6％(26/57) and 33.3％(19/57)] in the basic theory of basic care,knowledge and skills from the perspective of nursing targets and their relatives.The Chinese medicine nursing skills such as scrapping [needing very much 40.4％ (23/57)] and burying seeds [needing very much 40.4％ (23/57)] in the ear acupuncture point were also needed to be enhanced.A total of 75.4％ (43/57) of respondents chose online examination as the way of nursing personnel's evaluation.Conclusions Nursing personnel are in great need of on-job professional training.

Objective To analyze the efficacy and safety of transarterial chemoembolization (TACE) with chemotherapy regimen containing platinum in the treatment of primary liver cancer. Methods 86 patients with advanced hepatocellular carcinoma treated in our hospital from December 2015 to December 2016 were selected and randomly divided into two groups with 43 cases each group. Lipiodol chemotherapy was performed with Lobaplatin, epirubicin hydrochloride and mitomycin in the study group, control group while Lipiodol chemotherapy was performed with oxaliplatin, epirubicin hydrochloride and mitomycin in the control group. The gelatin sponge was used to block tumor blood vessels and the treatment lasted for 2 courses. According to the curative effect evaluation standard of solid tumor (RECIST), the curative effect was evaluated and the adverse reactions were compared between the two groups. Results The total effective rates of the two groups were not significantly different , but the effective rate of the study group (20.93%) was significantly higher than that of the control group (6.98%), and the difference was statistically significant (P<0.05). The incidence of hematologic toxicity in the study group (30.23%) was slightly higher than that in the control group (20.93%), the incidence of gastrointestinal reactions (44.19%) and liver and kidney damage (13.95%) were lower than that of the control group respectively by 60.47% and 25.58%, but there was no significant difference . The incidence of neurotoxicity in the study group (2.33%) was significantly lower than that in the control group (27.91%), and the difference was statistically significant (P<0.05). Conclusion For patients with advanced hepatocellular carcinoma, there was no significant difference between the efficacy and safety of transarterial chemoembolization with chemotherapy regimen containing platinum and oxaliplatin chemotherapy regimen, and adverse reactions are similar, but the incidence of neurotoxicity is low.

AIM and METHODS: To analysis the factor that involved in renal carcin ogenesis, we used the bait gene AK001518 to screen GenBank. To understand the re lationship between cell cycle related gene(CCRG) and p15, we did RT-PCR and Nort hern Blot experiments. Th en we examined CCRG expression level in renal carcinogenesis. RESULTS: Gained a function unknown gene CCRG that was 67% a mino acid identical with the gene AK00 1 518 that was regulated by p15. It was shown that the CCRG mRNA was dramatically d ecreased when p15 gene was over-expressed. CCRG expression level was much higher i n tumor tissues and cells than normal tissues and cells. CONCLUSION: The novel g ene CCRG expressed highly in the renal carcinoma, which might play a significant role in the renal carcinogenesis.

BACKGROUND: There are differences in physical and biological activity between the antibody from mammals and egg yolk antibody (IgY) from chicken. IgY is acid- and heat-resistant, and can prevent and cure the infectious diseases in animals and human being, which is also benefit to develop routine diagnostic immunoassays. Conventional ELISA assay for IgY takes much more time than dot-immunobinding assay.OBJECTIVE: To detect the IgY stability byusing dot-immunobinding assay.DESIGN: Open trail.SETTING: Department of Transfusion, Kunming General Hospital of Chinese PLA.MATERTALS: The experiment was completed in the Kunming General Hospital of Chengdu Military Area Command of Chinese PLA from January to June 2006. Two White Leghorn hens (30 weeks old) were selected. HLA-A*0201 α chain served as the antigen. The total protein concentration of the purified antigen was 0.04 g/L with the molecular mass of 32 000(self-prepared); nitrocellulose filter (NC, import and divided); nonfat dry milk (Anyi Corp. No. 20051220); DAB (Boshide Corp.);caprylic acid (made by Shanghai Xinghuo Chemical Factory); ammonium sulfate (Shantou Guanghua Chemical product).METHODS: ①HLA-A*0201 α chain with the total protein concentration of 0.04 g/L was purified with egg yolk antibody,and identified by SDS-PAGE. ②1 μL antigen was spotted into the center of NC membrane and dried in the incubator at 37 ℃. Then the NC membrane was blocked in 1 mL PBST and put in the incubator at 37℃ with shaking in 90 r per minute for 15 minutes. Then the liquid was exchanged with 1 mL PBST and added the primary antibody at a final concentration of 10 mg/L. After 30 minutes shaking in the incubator at 37 ℃, the NC membrane was washed in PBST for three times. The second antibody, mouse anti-chicken IgY conjugated to horseradish peroxidase (HRP) was added and after 30 minutes incubation, the NC membrane was washed three times in PBST. Binding was revealed by incubation with a DAB reagent. A positive reaction was represented by adeep brown spot,Irdlcating that IgY had better activity; if the spot became lighter IgY lost part activity, and when the spot disappeared, the IgY lost a the activty.According to intensity (gray degree)of the dot compared tothe standard, the remained percent of activity of the IgY was calculated. ③IgY was adjusted to three different protein concentrations with PBS: 1, 0.1, 0.01 g/L and stayed at room temperature for four months. 10 μg lgY was taken out from each concentration sample every month to detect the activity by dot-immunobinding assay. ④IgY was put into seven EP tubes with 100 μL per tube and numbered 1-7. Number 1 to 3 was adjusted pH to 5, 3 and 2, respectively with 1 mol/L HCI; Number 4 to 6 was to 9, 11 and 12, respectively with 1 mol/L NaOH. The pH of number 7 was neutral without adding acid or base. The samples were stayed in incubator at 37 ℃ for 3 hours. 10 μg IgY from each tube was taken every hour to detect the stability at different pH by dot-immunobinding assay. ⑤IgY was added to six EP tubes (10 μL per tube) and numbered 1-6. Number 1-6 was put into waterbath at 30, 40, 50, 60, 70 and 80 ℃ for 15 minutes. After cooled in refrigerator at 4 ℃, 10 mg samples from each tube and standard sample (untreated sample) taken to check the thermal stability by dot-immunobinding assay.MAIN OUTCOME MEASURES: ①SDS-PAGE of IgY. ②IgY stability at room temperature. ③IgY stability at different pH. ④ Detection of IgY thermal stability.RESULTS: ①Purified IgY after SDS-PAGE had two major binds, the molecular mass of the heavy chain was 66.000,and the light chain was 25 000. ②1, 0.1, 0.01 g/L IgY still had partial activity after staying at room temperature for four months. ③When pH ranged from 5 to 9, IgY still had partial activity after staying in 37 ℃ for 3 hours. If pH was lower than 5 or higher than 9, it lost the whole activity in above condition. ④Purified IgY was added to six EP tubes, the number 1-4 still had partial activity, but number 5 and 6 showed some white precipitate, which was caused by protein denaturation at higher temperature.CONCLUSION: IgY stability is higher than others. The dot-immunobinding assay described a rapid and simple method for the demonstration and characterization of functional activity of egg yolk antibody. With only small volume antigen and antibody, and specific dot, the dot-immunobinding assay method could process many samples at the same time.

OBJECTIVE:To establish the quality standards for Qiqilian capsule. METHODS:TLC was used to identify the As-tragali Radix,Phellodendri chinensis,Coptidis rhizom. HPLC was used to determine the contents of ginsenosides Rg1,ginsenosides Rb1 and notoginsenosides R1. The column was Shim-pack VP-ODS C18 with mobile phase of acetonitrile-water(gradient elution) at flow rate of 1.0 ml/min,detection wavelength was 203 nm,column temperature was 20 ℃. RESULTS:TLC of Astragali Radix,P. chinensis,C. rhizom showed cleer sports and good separation. The linear range was 0.9-9.0 μg for ginsenosides Rg1,0.94-9.4 μg for ginsenosides Rb1 and 0.3-3.0 μg for notoginsenosides R1(r≥0.999 5);RSDs of precision,reproducibility and stability test were lower than 3.0%;recoveries were 96.08%-99.75%(RSD=1.52,n=6),97.03%-99.75%(RSD=1.10,n=6)and 96.38%-98.55%(RSD=0.90,n=6),respectively. CONCLUSIONS:The method is simple,good reproducibility,and can be used for the quality control of Qiqilian capsule.

Objective To study the clinical effect of transpedicular vertebral osteotomy spine shortening in treating spinal kyphosis com-plicated with spinal cord nerve dysfunction. Methods A total of 80 patients with spinal kyphosis complicated with spinal cord nerve dys-function in our hospital from May 2013 to June 2014 were enrolled and randomly divided into observation group(n=40) and control group (n=40). The observation group received transpedicular vertebral osteotomy,and the control group received lamina and facet osteotomy. The situation of surgery,vertebral healing and spinal cord function condition,treatment effect between two groups were compared. Results The operation time and postoperative ambulation time of observation group were shorter than those of control group [(76. 52 ± 9. 1) vs (113. 46 ± 13. 44) min,(3. 28 ± 0. 43) vs (5. 67 ± 0. 68) d]. The postoperative bleeding volume,postoperative drainage volume of observation group were less than those of control group [(36. 14 ± 4. 28) vs (55. 23 ± 7. 15) mL,(17. 92 ± 2. 12) vs (29. 64 ± 4. 28) mL]. The Cobb angle and residual urine volume,initial and strong urinary bladder capacity,maximum urinary output of observation group were significantly less than those of control group [(6. 12 ± 0. 68) vs(9. 78 ± 1. 21) mL,(241. 45 ± 28. 56) vs(335. 54 ± 36. 86) mL,(456. 56 ± 51. 78) vs (586. 35 ± 63. 12) mL,(63. 78 ± 7. 24) vs (96. 32 ± 10. 22) mL]. The intervertebral height of observation group was higher than that of control group [(12. 62 ± 2. 81) vs (8. 41 ± 1. 32) mm]. The excellent rate of observation group was significantly higher than that of control group(97. 50%vs 82. 50%). Conclusion Transpedicular vertebral osteotomy spine shortening is helpful to reduce operation wound, pro-mote postoperative recovery,correct kyphotic deformity and improve neurological functionin,improve therapeutic effect.

Objective To improve the tri-primer allele gene amplification for realizing the single nucleotide polymorphisms (SNP) genotyping of the peripheral blood sample .Methods Aiming at the peripheral blood samples with the clinical usual antico-agulation processing by EDTA ,heparin and citrate ,with the locus rs1165205 as the target site ,the buffer solution(YW) suitable for whole blood was prepared and the PCR amplification system and the amplification condition were optimized for realizing the detec-tion of SNP genotyping .Results The genotyping results of locus rs1165205 by improved tri-primer allele gene amplification method were consistent with the results of the Sanger sequencing method ,and the peripheral blood samples treated by different anticoagu-lant were genotyped by the improved tri-primer ASA .Among 80 samples ,various genotypes of locus rs1165205 had no statistical differences in the distribution between the gout population and non-gout population(P= 0 .335) .Conclusion The improved tri-primer allele gene amplification method can be adopted to conduct the rapid genotyping research on gout SNP locus of the peripheral blood samples with the clinical usual anticoagulation processing .

Objective To establish the primary cat intestinal epithelial cells(IECs)culture methods and construct the cD-NA library for the following yeast two-hybrid experiment,so as to screen the virulence interaction factors among the final host. Methods The primary cat IECs were cultured by the tissue cultivation and combined digestion with collagenase XI and dispase I separately. Then the cat IECs cultured was identified with the morphological observation and cyto-keratin detection ,by using goat anti-cyto-keratin monoclonal antibodies. The mRNA of cat IECs was isolated and used as the template to synthesize the first strand cDNA by SMARTTM technology,and then the double-strand cDNAs were acquired by LD-PCR,which were subsequently cloned into the plasmid PGADT7-Rec to construct yeast two-hybrid cDNA library in the yeast strain Y187 by homologous recom-bination. Matchmaker?Insert Check PCR was used to detect the size distribution of cDNA fragments after the capacity calcula-tion of the cDNA library. Results The comparison of the two cultivation methods indicated that the combined digestion of colla-genase XI and dispase I was more effective than the tissue cultivation. The cat IECs system of continuous culture was established and the cat IECs with high purity were harvested for constructing the yeast two-hybrid cDNA library. The library contained 1.1× 106 independent clones. The titer was 2.8 × 109 cfu/ml. The size of inserted fragments was among 0.5-2.0 kb. Conclusion The yeast two-hybrid cDNA library of cat IECs meets the requirements of further screen research,and this study lays the foundation of screening the Toxoplasma gondii virulence interaction factors among the cDNA libraries of its final hosts.

Objective To evaluate the risk factors of septic shock after mini-percutaneous nephrolithotripsy (mPCNL). Methods Clinical data of 1 590 cases who underwent mPCNL from January 2013 to December 2014 were retrospectively analyzed. The x2 test and logistic regression were used to identify the key risk factors for septic shock after mPCNL. Results Of the 1 590 patients, 18 patients suffered septic shock, including 6 male patients and 12 fe﹣male patients. Their mean age was (45.6 ± 13.5) years (28 ~ 69 years). White cell in urine was 100 percent, the stone diameter ranged from 1.5 to 5.0 cm, unichannel for 15 cases while multichannel for 3 cases, the operation du﹣ration ranged from 45 to 200 min, mean (87.0 ± 56.0) min. 2 in 18 cases died in multiple organ failure, the others recovered till discharged. In x2 test, female gender (P = 0.001), (+++ ~ ++++) white cells in urine (P= 0.042), un-preoperative nephrostomy drainage (P=0.041) had significant association with septic shock after mPCNL. While in multivariate analysis, female gender ( O? = 5.471, 95 % CI: 0.756~21.452, P< 0.05) and un-preoperative nephrostomy drainage (O? =3.106, 95%CI:1.283~7.907, P<0.05) were identified as independent risk factors for septic shock after mPCNL. Conclusions Female gender and un-preoperative nephrostomy drainage are the key risk factors for septic shock after mPCNL.

ObjectiveTo evaluate the methods in treatment of upper urinary calculi in a low-head lithotomy with right or left lateral tilt by an ureteroscopy approach. Methods From September 2009 to May 2015, 110 patients with upper ureteral calculi (after failed ESWL) were underwent holmium laser lithotripsy by a ureteroscopy approach in a low-head lithotomy with right or left lateral tilt. Their clinical data and complications were analyzed retrospectively. Results Surgical effect of patients was satisfied with the success of gravel 91 patients, with a total rate of 82.7 %gravel. Conclusion Low-head lithotomy with right or left lateral tilt is a good body position to perform ureteroscopic lithotripsy for upper ureteral calculi. It is safe and effective.