Aim: To prepare chitosan nanoparticlescarrying gene and study its characteristics in vitro and in vivo.Methods:12-ACSs was dissolved in 0.05mol/L sodium acetate buffer to form a solution of 1mg/ml and a DNA(plasmid-encoding antisense ECE ) solution of 0.1mg/ml was dissolved in 25mmol/L Na2SO4.The charge ratio of components is selected as 2/1 for 12-ACSs /DNA complex.The complex was prepared by mixing 12-ACSs solution with DNA solution prior to observation by using transmission electron microscopy.Using Electrophoretic Retardation of DNA Migration detection,DNA precipitation,Binding Equilibration and DNase Resistivity Assay,the formation of 12-ACSs /DNA complex was determined and its stability was simultaneously evaluated.MTT Cell Proliferation Assays was performed on investigation of the cytotoxicity of 12-ACSs /DNA nanoparticles in bronchial epithelial cells (16HBE).The transfection efficiency of 12-ACSs /DNA nanoparticles in vitro and in vivo was investigated in 16HBE cells and Balb/c mice.Results: 12-ACSs /DNA nanoparticles (100~150nm) were observed by transmission electron microscopy.12-ACSs can protect the plasmid encoding antisence-ECE from degradation by DNase I.12-ACSs can transfer plasmid-encoding antisense ECE into 16HBE cells and into the airway epithelium in mice.As shown by fluorescent microscopy,green fluorescent protein reporter can be observed in the transfected cells as well as in the airway epithelium of the treated mice.And it showed a lower cell cytotocixity in cultured 16HBE cells and in mice treated with12-ACSs /DNA nanoparticles.Conclusion: In summery,the chitosan can be used as an effective protectant for DNA as well as an enhancer for in vitro gene transfection.

Abstract: Chigger mites belong to the phylum Arthropoda, class Arachnida, sub-class Acari, order Parasitiformes, family Trombiculidae and Leeuwenhoekiidae, with a wide range of species. There are more than 3 700 known species of mites globally, of which 531 species in 46 genera from three sub-families have been recorded in China, and 320 species of chigger mites recorded from 3 subfamilies and 32 genera in Yunnan Province. At this stage, chigger mites are the only vectors of scrub typhus, with about 60 species worldwide being potential vectors. Six vectorial mite species have been confirmed in China, including Leptotrombidium deliense, L.scutellare, L.rubellum, L.sialkotense, L.kaohuense, L.insulare. Yunnan Province has reported five vectorial mite species, including the previously-mentioned five species except for L.sialkotense. The zoogeographic study divided Yunnan Province into 5 zoogeographic areas, namely, Central Hengduan Mountains subregion, Southern Hengduan Mountains subregion, Eastern Yunnan Plateau subregion, Western Yunnan Plateau subregion, Southern Yunnan Mountainous subregion. The Southern Hengduan Mountains subregion has the highest number of known genera and species of scrub typhus and the highest number of scrub typhus cases, with 216 species in 22 genera from 3 subfamilies, and all 5 species of vector scrub typhus were distributed. Southern Yunnan Mountain subregion has the least number of mite species, with 91 species in 12 genera from three sub-families, with L. scutellare being the dominant vectorial mite species across the province. Due to suitable natural geographic conditions in Yunnan Province, vectorial mites can propagate rapidly, leading to a large number of scrub typhus cases. According to statistics, the number of scrub typhus cases in Yunnan Province has rapidly increased, with 15.40 times, 20.64 times and 28.91 times more cases in 2016, 2017 and 2018 respectively than in 2006. Scrub typhus remains a serious public health problem in Yunnan Province, posing a threat to the health of the local population. Therefore, prevention and control efforts should continue to focus on the affected areas and the general public.

Objective To evaluate the effect of replication protein A (RPA) gene suppression on the radiosensitivity of esophageal cancer cells (TE-1R ) and underlying mechanism. Methods A radioresistant human esophageal cancer cell line TE-1 R was screened out by fractionated irradiation to TE-1 cells,then siRPA1 or siRPa2 was transfected to TE-1R cells.The untransfected (Con) group and nonsense siRNA transfected (NC) group were set as control groups.The survival was measured with colony-forming assay and the cell cycle distribution was measured with flow eytometry.Results Compared with the Con and NC groups,the protein expression of RPA1 and RPA2 decreased significantly 48 h after siRPA1 and siRPA2 transfection.The D0,Dq,and SF2 values reduced from 2.09,1.70,0.85 in NC group to 1.67,0.71,0.44 and 1.82,0.89,0.51 in siRPAl and siRPA2 transfection groups,respectively.Accordingly,the sensitization enhancement ratios of Dq were 2.39 and 1.91,respectively.The G2/M arrest in siRPA1 and siRPA2 transfection groups increased from (18.701 3.14)％ of NC group to (26.95 ± 3.96)％ and (25.28 ± 2.74) ％ (t =2.827,2.853,P ＜0.05),respectively.Conclusions Knocking down of RPA1 or RPA2 genes can enhance the raidosensitivity of human esophageal cancer cells TE-1R,where the inhibition of radiation-induced sublethal damage repair may be involved.Accordingly,RPA may become a new target of radiosensitization in esophageal cancer.

Objective To compare the prognosis and life quality of patients with early-stage breast cancer(stage I or II)undergoing breast-conserving surgery or modified radical mastectomy.Methods 180 cases with early-stage breast cancer were divided into 2 groups:breast-conserving surgery group(BCS)and modified radical mastcctomy group(MRM),with 90 cases in each group.The 2 groups were all given radiotherapy,chemotherapy and endocrine therapy according to indications.The prognosis and life quality in the 2 groups were compared.Results The 5-year-postoperative follow-up showed there was no significant difference in duration of disease-free survival(DFS)of the 2 groups.However,life quality of BCS patients was significantly better than that of MRM patients in terms of general health,physical function,pain,vitality,social functioning,emotion and mental health.Conclusion Breast-conserving surgery is highly recommended for patients with early-stage breast cancer.

Objective To study the effect of low-level 84 disinfection solution on reproductive physiology of mice . Methods 72 KM mice were randomly divided into three groups :0.5%84 disinfection solution group ( group A) , 2.0%84 disinfection solution group (group B) and saline solution group (group C).12 males and 12 females in each group. Corresponding medicine was given once a day by intragastric gavage for 9 weeks.The estrous cycle of female mice was observed by exfoliative cytology of vagina .After 9 weeks, The sperm abnormality test was performed , Serum samples were taken for sex hormone test by ELISA, and the histological change of testis (or ovary), heart, liver, lung and kidney were observed.Results Compared with group C , No significant difference was observed in the weight variation and serum sex hormone level in group A and B (P >0.05).The difference of the estrous cycle of female mice was not significant (P >0.05).No histopathological abnormality was observed .Compared with group C, There was no significant difference in the rate of sperm abnormality in group A ( P >0.05 ) , but the rate in group B was significantly increased ( P <0.05 ) . Conclusion 0.5% 84 disinfection solution has no significant effect on reproductive physiology of mice , The concentration of disinfection solution must be strictly controlled when used .

Objective:To assess the vision-related quality of life(VRQoL) of the patients with simple vitreous hemorrhage after vitrectomy and analyze its influencing factors.Methods:A scale of life quality for patients with visual impairment(AQoL-DVI) were administered to 96 patients before and 6 months following vitrectomy.Scale scores were compared with vison preoperatively and postoperatively.Results:The lowest scores were at the item of "social life" and "spiritual and psychological status" before the surgery.The scores of the questionnaires increased significantly after vitrectomy.The change of visual acuity after surgery was a chief independent factor of the changes of questionnaire scores due to vitrectomy.Conclusion:The quality of life of patients with simple vitreous hemorrhage decreases sharply.VRQoL improves obviously after vitrectomy.

Background and purpose:Nowadays, about therapy of laryngeal carcinoma, people are paying more and more widely attention to ifnding out how to improve quality of patients’ life besides radical surgery. For glottic laryngeal carcinoma which invading anterior commissure or bilateral vocal cord, we performed modiifed frontolateral partial laryngectomy or modiifed cricohyoidoepiglottopexy, and contrastive analyzed the therapeutic efifcacy of the two ways. Methods:Sixty cases patients of glottic laryngeal carcinoma who treated in Hunan Provincial Tumor Hospital during 2005 to 2010, which invaded the anterior commissure or bilateral vocal cord, were randomly attributed to two groups as A and B;30 patients of group A were underwent modiifed frontolateral partial laryngectomy and repaired with bilateral sternohyoid muscle lfap, 30 patients of group B were treated by modiifed cricohyoidoepiglottopexy. Follow-up time of each patient was 5 years postoperation and clinical data were retrospectively analyzed. Results:The 5-year survival rate was 86.7%in group A as well as 83.3%in group B, and there was no statistical difference between two groups (P=0.718). Pronunciation function:22 cases in group A and 21 cases in group B whose pronunciation function can be competent in the noisy environment, and can pronounce“a”and“i”vowel;8 cases in group A and 9 cases in group B can pronounce only a“ha”,“hi”sound, that couldn’t communicate with others in a noisy environment. There was no statistical difference in pronunciation function between the two groups (P=0.774). Incidence of deglutition disorder 4 weeks postoperation:group A was 0 (0/30), group B was 16.7%(5/30), and the difference between the two groups was statistically signiifcant (P=0.026);average time of extubation postoperation:group A was (10±2.3) d, group B was (20±4.6)d, and the difference between the two groups was statistically signiifcant (P=0.0000);recurrence rate of dyspnea after extubation:group A was 16.7%(5/30), group B was 0 (0/30), and there was statistically signiifcant difference between the two groups (P=0.026). Conclusion: For the glottic laryngeal carcinoma which invading anterior commissure or bilateral vocal cord, there was no statistical difference in 5-years survival rate and function of pronunciation between modified frontolateral partial laryngenctomy and modified cricohyoidoepiglottopexy postoperation. The former had less postoperative deglution disorder, earlier extubation time, and to some extent, alleviated the suffering of the patients, but part of these patients needed secondary surgery due to dyspnea which resulted by radioactive tissue adhesion after extubation. The latter had more serious deglution disorder postoperation, longer recovery time, and relatively longer time to extubating, showed no again dyspnea after extubation, and had more extensive adaptation disease. In a word, each way of operation has its advantage respectively.

Objective To evaluate the effectiveness of discharge planning and home care for patients with peritoneal dialysis (PD).Methods A total of 60 PD patients were randomly divided into the intervention group and control group,30 cases in each group.Discharge planning and home care were carried out in the intervention group, while routine nursing and telephone follow -up service after discharge were implemented in the control group.The patientsˊscores of Zung Self -Rating Anxiety Scale(SAS),Self -Rating Depression Scale(SDS),Self management ability of patients undergoing peritoneal dialysis,complication rate and readmission rate were compared between the two groups at the next day admission,discharge,lth month and 3th after discharge.Results The scores of SAS and SDS in the intervention group were significantly lower than that in the control group at 3th month after discharge,the difference was statistically significant(tSDS =5.263,tSAS =3.812,P <0.05).The Self management ability of patients undergoing peritoneal dialysis was significantly higher than that of the control group at lth and 3th month after discharge,the difference was statistically significant (t =6.845,t =7.231,P <0.05).No significant difference in the re -admission rate (16.67%,6.67%)was found between the two groups (χ2 =0.387,P >0.05).The complication rate in the intervention group was significantly lower than in the control group at 3th month after discharge(χ2 =5.124,P <0.05).Conclusion Discharge planning and home care can effectively relieve depression and anxiety in patients with peritoneal dialysis and improve the abilities of daily life.It is worth promoting and applying in clinic.

Objective To realize the actuality and analyze the problem of the hospital medicine preparation in Beijing,then to discuss its developing methods.Methods By summarizing questionnaire of the hospital medicine preparation and combining with the problem showed in daily supervision,we analyzed the main existing problems and reasons for hospital medicine preparation and put forward the developing methods.Results The main existing problems of hospital preparation are the decreasing variety of medicine preparation,insufficient infrastructure and software.Conclusion Therefore we should enhance the input on the infrastructure,set up the software,improve the quality of the staff,and consummate the regulation means and system.

Background Krüppel-like factor 6 (KLF6) is related to the physiological or pathological process,such as growth,cell differentiation,proliferation,apoptosis,angiogenesis,tissue repair,and so on.But in ophthalmology,it is less reported about the expressing level of KLF6 protein in lens epithelial cclls (LECs) or the effect of KLF6 on the proliferation of human LECs.Objective This study was to investigate whether KLF6 can inhibit proliferation of human LECs.Methods KLF6 eukaryotic expression plasmid (pEGFP-C2-KLF6) was constructed using reverse transcription PCR(RT-PCR) and identified by double enzyme digestion method and PCR.Human LECs strain (HLE-B3) was cultured and passaged using low glucose DMEM containing 10％ fetal bovine serum and then divided into 4 groups.KLE-B3 transfection reagents were added in the culture medium of all groups.In addition,no agent was used in the blank control group;only insulin-like growth factor-1 (IGF-1) was appended to the medium in only IGF-1 group ;null vector was transfected and IGF-1 was appended in the null plasmid transfection+ IGF-1 group;while pEGFP-C2-KLF6 eukaryotic expression plasmid was transfected into the cells,and simultaneously IGF-1 was added in the pEGFP-C2-KLF6 plasmid transfection+IGF-1 group.After 24 hours of intervene,water soluble tetrazolium salt-1 (WST-1) test was used to detect the growth status of the cells,and Western blot assay was used to assay the relative expressing level of KLF6 protein in the cells.In the other hand,the cells were cultured at the density of 1 ×104/piece,and 0,0.10 and 0.25 μg pEGFP-KLF6 were transfected into each piece of cells respectively,and then IGF-1 was added with a final concentration of 50 μg/L for 24 hours after cell culture.Expressions of Ki-67 protein and mRNA in the cell pieces were detected by immunocytochemistry and fluorescent semiquantitative PCR,respectively.Results The PCR product bands were consistent with KLF-6 gene in length,and the product fragments were corresponding with expectant ones via PCR and double enzyme digestion method,showing a successful construction of pEGFP-C2-KLF6 eukaryotic expression plasmid.After 24 hours of IGF-1 stimulation,the absorbance values of the cells were 0.86±0.00,2.10±0.01,2.24±0.12 and 1.06±0.02 in the blank control group,only IGF-1 group,null plasmid transfection + IGF-1 group and the pEGFP-C2-KLF6 plasmid transfection + IGF-1 group,with a significant difference among the 4 groups (F =38.322,P ＜ 0.05),and that in the pEGFP-C2-KLF6 plasmid transfection +IGF-1 group was significantly lowed in comparison with the only IGF-1 group and the null plasmid transfection+IGF-1 group (q=6.42,7.31,both at P＜0.05).Western blot assay showed that the relative expressing levels of KLF6 protein were statistically different among the four groups (F =591.858,P＜0.05),and those in the pEGFP-C2-KLF6 plasmid transfection+IGF-1 group were 1.47,2.04,3.27 folds higher than those in the blank control group,only IGF-1 group,respectively and null plssmid transfection+IGF-1 group,respectively.Immunocytochemistry revealed that the expressing intensity of Ki-67 protein was gradually weakened with the decrease of pEGFP-C2-KLF6 plasmid dose in the 0,0.10 and 0.25 μg pEGFP-C2-KLF6 plasmid transfection+IGF-1 groups.Fluorescence semiquantitative PCR results showed that the relative expression values of Ki-67 mRNA in the cells were O.15±0.08 and 0.11±0.03 in the 0.10 μg and 0.25 μg pEGFP-C2-KLF6 plasmid transfection+ IGF-1 groups,which were significantly lower than O.77± 0.12 of the 0 μg pEGFP-C2-KLF6 plasmid transfection group,with a statistically significant difference among the three groups (F =54.825,P＜0.05).Conclusions KLF-6 can effectively inhibit IGF-1-induced proliferation of human LECs,and it can be regarded as one of the regulatory factors of the proliferation of HLE-B3 cells.