AIM:To observe whether modified epitopes from osteosarcoma high-expressing antigen papilloma-virus-binding factor (PBF) have HLA-A2 restricted antitumor ability,and to develop peptide-based immunotherapy for os-teosarcoma. METHODS:RT-PCR and Western blot were used to determine the expression of PBF in the osteosarcoma cell lines U2OS and Saos-2. HLA-A2 epitopes from PBF protein were predicted by NetCTL 1.2, SYFPEITHI and IEDB. The modified peptides from PBF containing HLA-A2 binding anchor motifs were designed by replacing the anchor residues. The peptides were synthesized by standard solid-phase methods,and the binding affinity of the peptides to HLA-A?0201 was evaluated by T2A2 cell binding assay. ELISPOT assay was used to investigate the seretion of interferon-γ(IFN-γ) from the peptide-induced specific cytotoxic T-lymphocytes (CTLs). The ability of inducing T-cell response was analyzed by lactate dehydrogenase (LDH) release assay and carboxyfluorescein succinimidyl ester (CFSE) cytotoxicity assay in vitro. RE-SULTS:The expression of PBF was observed in the U2OS and Saos-2 cells. The candidate peptides P75-1Y2L,P412-1Y, P416-1Y2L9V, P107-1Y and P435-1Y2L showed moderate affinity toward HLA-A2 molecule. The modified peptides showed significantly higher affinity with HLA-A2 than the native peptide. ELISPOT assay showed that P412, P412-1Y, P416,P416-1Y2L9V and P435-1Y2L induced specific CTLs to secrete IFN-γ, and P412-1Y and P416-1Y2L9V induced more secretion of IFN-γ than the native peptide. The CTLs induced by P412, P412-1Y, P416 and P416-1Y2L9V lysed U2OS cells. P412-1Y and P416-1Y2L9V peptide-specific CTLs showed higher cytotoxicity against U2OS cells than the native peptide-specific CTLs. CONCLUSION:Compared with the native peptide,modified epitopes P412-1Y and P416-1Y2L9V have higher binding affinity with HLA-A?0201 and retain immunogenecity. In addition,the anti-tumor immunity effects of modified epitopes P412-1Y and P416-1Y2L9V are stronger than the native peptide. The peptides P412-1Y and P416-1Y2L9V is excellent HLA-A?0201 restricted CTL epitopes from tumor antigen PBF,which could serve as new can-didates towards antitumor peptide vaccines.

Objective To study the anemia status and genotype of thalassemia in preschool children in Shenzhen. Methods 658 preschool with anemia hospitalized in Shenzhen Longgang Maternal and Child Health Hospital from October 2012 to September 2015 were screened by complete blood analysis . The most common mutations of thalassemia genotype (17 β thalassemia genotype mutation,3 α thalassemia genotype mutation and 3α thalassemia genotype absence change) in Chinese population were detected. Results All cases have microcytic hypochromic anemia. 426 cases were identified to be thalassemia (64.7%). 23 genotypes and 13 gene mutation type were detected. The most common genotype type were SEA/αα(46%),β654/βN(15%),β41?42/βN(12.7%). And the most common allele gene mutation type were SEA (49.1%),IVS?Ⅱ?654( C→T)(14.4%),CD41?42(?TTCT)(12.4%) re?spectively. MCV and MCH of thalassemia children was significantly lower than that of children diagnosed as without thalassemia. The differences of RBC,Hb,MCV,MCH,MCHC,RBC?SD between the two groups were statistical?ly significant. Conclusion The proportion of thalassemia among preschool anemia children in Shenzhen area was high,and it is necessary to strengthen the local thalassemia prevention and decrease anemia effect on preschool children′s health.

Objective To evaluate the killing effects of oxymatrine(OM) on human colon cancer cell line SW1116 and the mechanism of its anti-neoplastic effect. Methods Methyl thiazolyl tetrazolium analysis, flow cytometry , polymerase chain reaction(PCR)-enzyme linked immunosorbent assay (ELISA) and RT-PCR were used to detect the killing effects of OM and its influence on cell cycle dis- tribution , telomerase activity, expressions of human telomerase reverse transcriptase (hTERT), c-myc ,p53 and mad1 genes in SW1116 cells. Results OM exhibited dose-dependent killing effects on SW1116 cells and induced the increase of G1/G0-phase cells and decrease of S-phase cells. It was found that OM could supress the telomerase activity of SW1116 cells, and the effects were dose- and time-dependent. After OM administration, the expression of hTERT gene in SW1116 cells was decreased, those of p53 and mad1 genes were increased, and the expression of c-myc gene had no marked changes. Conclusions OM has dose-dependent killing effects on SW1116 cells.The anti-neoplastic activity of OM might be due to the inhibition of telomerase activity by means of its influence on hTERT and the up-stream regulation genes.

Objective This study was designed to determine the clinical pharmacokinetics ofcisatracurium in Chinese patients. The purpose of this study was twofold: the determination of ED50 and ED95 of cisatracurium and the intubating conditions and changes in BP and HR produced by 2×ED95 and 3 ×ED95 . Methods 100 patients scheduled for elective surgery, aged 30-65 years, served as the experimental subjects. Anesthesia was induced with propofol 2mg/kg and fentanyl 4μg/kg. 5 min after BP and HR were stabilized the study was started. The study consisted of two parts according to the purpose of the study. In part Ⅰ 70 patients were included and randomized to 7 groups based on the doses of cisatracurium the patients received. The doses were 25, 30, 35, 40, 45, 50 and 55 μg/kg from group Ⅰ to Ⅶ. Neuromuscular transmission was assessed by measuring the responses of adductor pollicis muscle to stimulation of ulnar nerve with TOF pulses.The maximal depression of Tt was measured after administration of cisrtracurium, and ED50 and ED95 were calculated. In part Ⅱ 30 patients were included and divided into two groups to receive 2×ED95 (group Ⅷ ) or 3×ED95 (group Ⅸ ) of cisatracurium. The intubating conditions were evaluated. The onset time and time of recovery from paralysis as well as the changes in BP and HR after administration of cisatracurium were recorded. ResultsED50 and ED95 for cisatracurium were 30.7μg/kg and 53.4μg/kg respectively. The onset times for 2×ED95 and 3×ED95 were (254±84)s and (190±59)s. The durations of action was (38.8±6.1)min and (53.28.8)min respectively. Recovery indexes were (16.4±4.9) min and (15.6±5.2) min. There were little changes in BP and HR after administration of cisatracurium in both group Ⅷ and Ⅸ. The intubating conditions were not significantly different. ConclusionsED50 and ED95 for cisatracurium are 30.7μg/kg and 53.4μg/kg in Chinese patients. The onset time is shorter and duration of action longer with 3×ED95 which does not significantly affect BP and HR.

Objective:To analyze and summarize the commonalities and patterns in acupoint selection for moxibustion treatment of asthma. Methods: Data retrieval was conducted using ‘moxibustion’ and ‘asthma’ as the keywords through China National Knowledge Infrastructure (CNKI), Wanfang Academic Journal Full-text Database (Wanfang) and Chongqing VIP Database (CQVIP). Excel 2010 was used to establish the major acupoint database for moxibustion prescriptions in treatment of asthma; data mining methods including association patterns and clustering were adopted to analyze the characteristics and patterns in acupoint selection for moxibustion treatment of asthma. Results: A total of 161 moxibustion prescriptions were recruited. The most commonly used acupoint was Feishu (BL 13), the most commonly used meridian was the Bladder Meridian of Foot Taiyang, and the most commonly treated region was the back. The association rule analysis showed that Feishu (BL 13)-Dazhui (GV 14)-Fengmen (BL 12) had the most significant correlation, and the clustering analysis discovered 5 effective acupoint clusters. Conclusion: In moxibustion treatment of asthma, topical acupoints Feishu (BL 13), Dazhui (GV 14) and Fengmen (BL 12) can be selected, along with the acupoints from the Bladder Meridian of Foot Taiyang and the back.

Objective To evaluate the clinical value of pulmonary embolism severity index (PESI) in non high-risk acute pulmonary thromboembolism (APTE) patients treated with sequential anticoagulation.Methods Non high-risk APTE patients treated with sequential anticoagulation were divided into two groups according to PESI:high-value group and low-value group.Prognosis and treatment response was compared between two groups.Results There were 82 cases in high-value group,and 76 cases in low-value group.The rate of adverse events in high-value group was significantly higher than that in low-value group [23.2％(19/82) vs.7.9％ (6/76)] (x2 =5.0698,P =0.009),and 30 days cumulative hazard was also significantly higher than that in low-value group (P ＜ 0.05).The sensitivity of predicting adverse events by PESI was 76.0％,specificity was 52.6％,positive predicting value was 64.6％,and negative predicting value was 65.9％.The mortality in high-value group was significantly higher than that in low-value group [9.8％(8/82) vs.1.3％ (1/76)] (P =0.022).After 30 days of anticoagulation,the pulmonary artery systolic pressure,internal diameter of right ventricle in high-value group was significantly higher than that in low-value group [(39.4 ± 8.1) mm Hg (1 mm Hg =0.133 kPa) vs.(27.2 ± 5.5) mm Hg,(33.0 ± 7.8) mm vs.(21.7 ± 4.6) mm] (P =0.034,0.021),and arterial oxygen partial pressure was significantly lower than that in low-value group[(75.15 ± 12.41) mm Hg vs.(86.36 ± 9.22) mm Hg](P=0.016).Conclusions PESI can effectively predict short-term prognosis of non high-risk APTE patients treated with sequential anticoagulation.At least some of these patients might need treatment other than sequential anticoagulation.

Objective To evaluate efficacy of routine cleaning and disinfection methods for incubators,and put forward a feasible improvement solution.Methods 30 incubators used in a neonatal intensive care unit of a hospital between Decem-ber 2013 and June 2014 were chosen and randomly divided into baseline,control,and trial groups(10 incubators in each group).Baseline group and control group were disinfected by routing disinfection method (wiping internal and external sur-faces of incubators with water and chlorine-containing disinfectant),trial group adopted intensified disinfection method (wi-ping internal surfaces of incubators with alcohol)on the basis of routine disinfection,disinfectant efficacy of three groups were compared.Results In baseline group,unqualified incubators were initially detected on the fourth day of monitoring, all incubators were contaminated in varying degrees on the seventh day of monitoring,the detection rate of unqualified spec-imens was 31.43% (88/280).The median time for the initial detection of unqualified incubators in control group and trial group were on the fifth day and seventh day respectively,there was significant difference between two groups(χ2 =12.38, P <0.05);The unqualified rate of trial group was significantly lower than control group (15.36%[43/280]vs 32.86%[92/280],χ2 =23.43,P <0.05 ).Conclusion Intensified disinfection with alcohol on the basis of routine disinfection method can effectively improve the disinfectant efficacy of the surface of incubators,it is convenient,inexpensive and safe, and worth to be popularized in primary hospitals.

Objective To observe the clinical curative effect of thoracic dorsal root ganglion pulse radio frequency stimulation combined with pregabalin in treatment of postherpetic neuralgia (PHN).Methods Sixty-two PHN patients were randomly divided into the single drug group and combination group (n=31).The pre-and post-treatment VAS values and pittsburgh sleep quality in-dex (PSQI),decreased dosage of tramadol after treatment,adverse reaction during treatment,and long-term therapeutic effects were observed.Results VAS values,PSQI and tramadol dosage de-creased significantly post-treatment in both groups,especially in the combination group (P < 0.05). In addition,the incidence of dizziness,somnolence and ataxia was less,but puncture pain was more in combination group (P < 0.05 ).Long-term therapeutic effects of patients in the combination group were significantly improved.Conclusion Thoracic dorsal root ganglion pulsed radio frequence com-bined with pregabalin is effective in the treatment of PHN.

Objective To investigate the epidemiological characteristics of the group A rotavirus (RV) among the children with diarrhea in Shenzhen ,in order to provide reference for clinical diagnosis and treatment .Methods Stool samples were collected for RV detection from children with diarrhea .The results were analyzed .Results A total of 3 509 cases of children with RV infection were checked out from 14 511 cases of children with diarrhea ,with the infection rate accounting for 24 .18% .RV infection occurred all year around ,and the infection peak was November ,December and January .RV infected children mainly distributed in the age group of 6 months to 2 years .Conclusion RV infection in children is a universal problem in Shenzhen .Clinic should pay attention to it .

The stability of the intraocular lens ( IOL ) after cataract surgery is composed of decentration, tilt, rotation, and the change of anterior chamber depth. Its stability is an important factor affecting postoperative visual quality. By analyzing the related factors which influence the stability of intraocular lens, improvements can be identified for future cataract operations. The stability of intraocular lens is influenced by many factors: intraocular structure, the size and the symmetry of intraoperative capsulorhexis, the position of the intraocular lens, the material and design of the intraocular lens, etc. In order to improve the patient's vision, cataract surgeries have been experiencing an evolution. IOL material have also been contributing to such innovations.