Objective:To construct a phage display antibody library of special antigen responding to serum starved U251 cell,from which the serum responding gene and protein would be gotten.Methods:U251 cells were cultured in serum-absent midium for 48 h.Its protein was extracted and used to immunize BALB/c mice.Total RNA of the spleenocytes of immunized mice was extracted.VH and VL were amplified by RT-PCR and were linked into ScFv(Single chain fragment of variation) with a linker.ScFv was recombined to pCANTAB5E vector and was transformed to TG1 strain.Results:The library capacity was up to 3?106 cfu/L.A positive clone was identified from 8 random clones of this library.Conclusion:The special ScFv phage library is constructed successfully.It is the basis for screening special antibodies which can recognize serum responding protein.

Objective To explore the changes of PH values of N,O-CMC/β-TCP compositive materials with different mass fractions in simulated body fluids (SBF)and their influence in the growth of MG63 cells, and to illustrate their mechanisms, and to provide reference for the further research on the bone repair materials. Methods The N,O-CMC/β-TCP with mass fractions of 2/1,1/1 and 1/2 were used as experimental groups,and the collagen nano calcium phosphate bone repair material as control group. The materials with different mass fractions were immersed in SBF and the pH values were measured by pH meter after soaking for 7,14,21 and 28d,respectively.The MG63 cells with the concentration of 1 × 105 mL-1 were inoculated and co-cultured in experimental and control groups,the adhesion and morphological changes of MG63 cells in each group were observed by scanning electron microscope and the cell proliferation was detected by MTT method after co-culturing for 2,4 and 6 d.Results The pH values were 6.70-7.25 in N,O-CMC/β-TCP (1/2)group and N,O-CMC/β-TCP (2/1)groups and the pH value in N,O-CMC/β-TCP (1/1)group was basically 7.15. The cells in N,O-CMC/β-TCP (2/1)group formed owe full,spreading face small and less secretion,but the cells in N,O-CMC/β-TCP 1/2 and 1/1 groups formed in full, pseudopodia interconnection, widely spreading and more secretions under electron microscope. The proliferation rate of the cells in N,O-CMC/β-TCP with (1/1 ) and N,O-CMC/β-TCP (2/1)groups had no statistical difference compared with control group (P>0.05),but there was significant difference between control group and N,O-CMC/β-TCP (1/2)group (P<0.05).Conclusion The changes of pH values of N,O-CMC/β-TCP materials with different mass fractions in SBF are small and the pH values are neutral;the order of the mass fraction of N,O-CMC/β-TCP to promote the growth of MG63 cells is 1/1,2/1,and 1/2.

Objective To evaluate the clinical efficacy and safety of bevacizumab combined with chemotherapy in the treatment of recurrence ovarian cancer.Methods Thirty seven ovarian cancer patients with complete data who treated with bevacizumab combined with chemotherapy were reviewed from the data bases of our hospital and included in this retrospective study.All included patients received more than 2 cycles of bevacizumab based combination therapy with eligible for the efficacy and toxicity assessments.The objective response rate was recorded and evaluated according to the response evaluation criteria in solid tumors (RECIST) and cancer antigen 125 (CA125) criteria.The data of survival time and adverse drug reactions were analysised . Results After treatment, the objective response rate were 32.4%and 43.2%respective-ly by the RECIST criteria and CA125 criteria.The median progression free survival for the 37 cases was 5.4 months.The two complete response patients had longest DFS interval with one for 10.0 months and another for 11.2 months.The main hematologic toxicity and non-hematologic toxicity were gradeⅠ-Ⅱneutropenia, thrombocytopenia, nausea and vomiting et al.Conclusion With acceptable toxicity and relative well clinical activity, the bevacizumab combined chemotherapy can be a potential treatment modality for the recurrent or platinum-refractory ovarian cancer.

Objective To compare the changes of diopter and axial length(AL) m 2 years after wearing orthokeratology lens,rigid gas permeable contact lens (RGPCL) and spectacles in adolescent myopia,and evaluate the control effects of different methods on adolescent myopia.Methods Prospective clinical study.The date of 99 eyes of 50 adolescent myopia cases wearing orthokeratology,95 eyes of 48 adolescent myopia cases wearing RGPCL,and 100 eyes of 50 adolescent myopia cases wearing full correction single vision spectacles were collected from June 2012 to June 2014.The follow-up time was 2 years.The status of diopter and AL before and wore mirror after 2 years and the uncorrected visual acuity (UCVA) in group of orthokeratology were recorded.The examinations were repeated after 2 years without wearing orthokeratology and RGPCL at least 30 days.Results There was no statistical difference in age,refractive status and AL among three groups before wearing the mirror.After wearing for 2 years,AL and spherical equivalent in three groups were increased (F =278.43,140.12;all P ＜ 0.01).The increase of AL and SE was as follows:orthokeratology group (0.30 ± 0.39) mm,(0.83 ±0.97) D;RGPCL group (0.55 ± 0.41) mm,(1.55 ± 0.63) D;spectacles group (0.59 ± 0.34)mm,(1.73 ± 0.62)D.The increase of orthokeratology group was the least,and there was no statistical difference between the other two groups.UCVA was significantly improved at each time point after wearing orthokeratology(F =486.38,P ＜0.01).Conclusion In the three optical correction methods of orthokeratology,RGP and spectacles,the ortho-keratology is the best on controlling adolescent myopia,and no difference between the other two.At the same time,wearing orthokeratology can also significantly improve UCVA in myooia cases.

Objective: To construct the public health performance evaluation index system for the county medical community, so as to provide reference for the assessment of the public health work in the county medical community. Methods: According to the 2019 Public Health Tasks of Zhejiang Province and the current status of the county medical community's public health work, a framework was developed. Twenty experts from universities, CDCs, and county medical community were invited to construct the index system after two rounds of Delphi expert consultation, and the index weight was determined by the analytic hierarchy process. Results : The experts aged ( 46.10±6.41 ) years and worked for ( 23.85±7.37 ) years, all of whom had a bachelor degree or above, and 18 had associate senior titles or above. The response rates of two rounds of consultation was both 100.00%; the authority coefficient was 0.811; the values of Kendall's W all had statistical significance ( P<0.05 ) , which in the second round were higher than those in the first round. The public health performance evaluation index system for county medical community finally included 10 first-level indexes, among which priority diseases surveillance and management weighed most ( 0.165 7 ) ; 32 second-level indexes, among which food-borne diseases surveillance, infectious diseases/public health emergencies reporting and infectious diseases/public health emergencies response weighed the top three ( 0.071 5, 0.064 7, 0.063 8 ); 120 third-level indexes, among which the timely reporting rate of food safety incidents, the reporting rate of infectious diseases and the information response rate of infectious diseases/public health emergencies weighed the top three ( 0.039 4, 0.022 1, 0.021 7 ). Conclusion The public health performance evaluation index system of the county medical community has been established, which can provide reference for the health administrative departments.

BACKGROUND: The immature dendritic cell (imDC) can induce immunological tolerance and has widely application in the field of organ transplant. At present, the methods of inducing imDC are insufficient, so the new induction method is demanding.OBJECTIVE: To investigate the effect of sodium butyrate (SB) on the maturation and immunological function of human peripheral blood-derived imDC.DESIGN: Controlled observation and in vitro cytological trial.SETTING: Department of Hepatobiliary Surgery in the Second Affiliated Hospital of Sun Yat-sen University.MATERIALS: Five samples of human peripheral blood were obtained from the healthy volunteers (aged 20-23 years) of Sun Yat-sen University, totally 500 mL. Then peripheral blood mononuclear cells (PBMCs) and lymphocytes were isolated within 2 hours.METHODS: The experiment was carried out in the Medical Research Center of the Second Hospital Affiliated to Sun (1 mmol/L) was added for induction, while those supplemented with maturation promoting factor lipopolysaccharide (LPS)the beginning of induction, while LPS was added on the sixth day for second stimulation.MAIN OUTCOME MEASURES: Cell morphological change, flow cytometry was used to detect DC phenotype,FITC-labeled Dextran was used to detect the endocytosis of DC, the production of IL-12 was determined by means of enzyme-linked immunosorbent assay, and the proliferation of lymphocyte induced by DC was assayed with mixed lymphocyte reaction.expressions of CD80, CD83 and HLA-DR were significantly lower in the imDC of routine induction group following SB maturity promoting, compared with LPS group (P＜0.01). On the sixth day, LPS was added into the SB-induced imDC,Endocytosis of DC: The imDC of routine induction group possessed a significantly lower endocytic activity after induced by LPS, and there were extremely significant differences compared with blank control group and SB maturation Production of IL-12: The production of IL-12 in the mDC induced by LPS was significantly higher than that in control group, SB maturation promoting group and SB induction group, the mDC induced by LPS in routine induction group stimulated significantly stronger proliferation of lymphocyte (P＜0.01).

AIM: To investigate the immunological function of sodium butyrate-induced immature dendritic cells in vitro.METHODS: The human monocyte-derived dendritic cells were induced in the presence of human granulocyte macrophage-colony stimulating factor(GM-CSF) and interleukin-4 (IL-4), combined with sodium butyrate. The immunological function of sodium butyrate-induced dendritic cells was detected by the FCM, endocytic activity, T cells stimulatory proliferation capacity, and interleukin-12 (IL-12) production.RESULTS: Sodium butyrate could down-regulate the major histocompatibility complex(MHC) class II and costimulatory molecules of dendritic cells, increase the endocytic activity, induce a stage of T-cell anergey, and inhibit the T helper cell type 1-skewing factor IL-12 production. CONCLUSION: Sodium butyrate inhibits the maturation of dendritic cells and induces production of immature dendritic cells, which may help to explore the machenism of its epigenitic modification.

OBJECTIVETo investigate the biological characteristics of xenogenic acellular dermal matrix( Xeno-ADM) incorporated with silver, and to observe its effect in grafting.

METHODSXeno-ADM was prepared with 0. 25% trypsin and 0. 5% triton X-100 , and then it was immersed in 2 g/L silver nitrate solution to prepare xeno-ADM incorporated with silver. The bacterial inhibitory effect of two kinds of xeno-ADM on burn wound was determined, and the histological characteristic of the wounds was observed with optical microscope, transmission electron microscope and scanning electron microscope. The Ag+ content in the xeno-ADM incorporated with silver was measured with atomic absorption spectrometer. Twenty-seven rabbits with full-thickness skin defects on the back were randomly divided into three groups, i. e. split-thickness skin autograft only( group A, n = 9) , xeno-ADM with overlying split-thickness skin autograft( group B, n = 9) , xeno-ADM incorporated with silver with overlying split-thickness skin autograft ( group C, n = 9). The skin specimens from grafted area in each group were harvested at 2,4 and 6 post-operation weeks( POW) and examined under light microscope and transmission electron microscope. The condition of the graft, the contraction degree of the grafts and the historical changes in grafting area were observed at 2,4,6 POW. The survival rate of the grafts was calculated and the proliferative activity of the lymphocyte in each group was determined at 2 POW.

RESULTS1. Compared with xeno-ADM, the anti-bacterial effect of xeno-ADM incorporated with silver was much better ( P < 0. 05). No epidermis was seen in both types of xenografts ,and the collagen fibers were even in size and arranged regularly, with no obvious degeneration, and the dermis was also devoid of cells and cellular components. The Ag + content in xeno-ADM incorporated with silver measured (2. 7+/-0. 7) mg/g. 2. The grafts in B and C groups presented similar color to that of normal skin at 6 POW, and it was smooth, with fine texture and no scarring. The collagen fibers was arranged regularly, and conjunction between epidermis and dermis, the structure of basal cell desmosome and semi-desmosome were well reconstructed. The grafts in A group was in dark red color, with obvious contraction, and easily broken. The contraction rate in A group at 2,4 and 6 POW were obviously higher than those in B and C groups( P < 0. 05), while no obvious difference was observed between B and C groups. ( P >0. 05). The overall survival rate of the grafts in C group at 2 POW was 91.7% , which was evidently higher than that in A (77.8%) and B (80. 6% ) groups. The lymphocyte proliferative activity exhibited no difference among A, B and C groups( P >0.05).

CONCLUSIONThe xeno-ADM incorporated with silver has good anti-bacterial effect. In addition, it preserves the basic tissue structure and integral collagen fiber scaffold, without cells to induce rejection, so that it can be used as an ideal dermal substitute.

Animals ; Dermis ; transplantation ; ultrastructure ; Rabbits ; Silver ; therapeutic use ; Skin Transplantation ; Skin, Artificial ; Swine ; Transplantation, Heterologous ; Wound Healing

OBJECTIVETo study the pathogenic bacteria of lower respiratory tract infection (LRTI), and age and gender distribution and drug resistance of the pathogenic bacteria in children.

METHODSSputum specimens for bacterial cultures were collected in sterile tubes from all of the children with LRTI who had been admitted to the Children's Hospital of Zhejiang University between August 2001 and July 2002. Antibiotic susceptibility tests were performed using the Vitek system, the Kirby-Bauer diffuse method and the Etest method after bacteria were identified.

RESULTSAmong the 4,238 patients with LRTI during the study period, 1,181 patients were bacteria-positive, with a positive rate of 27.9%. Streptococcus pneumoniae (S. pneumoniae) was the most common (222 strains), followed by Haemophilus influenzae (H. influenzae) (216 strains), Klebsiella pneumoniae (K. pneumoniae) (216 strains), Escherichia coil (E. coli) (169 strains) and Staphylococcus aureus (S. aureus) (89 strains). The isolation rate of S. pneumoniae in females was significantly higher than in males (6.2% vs 4.7%; P < 0.05). However, the isolation rates of K. pneumoniae and S. aureus in males were higher than in females (5.1% vs 4.1% and 2.5% vs 1.5%, respectively; P < 0.05). A higher incidence of LRTI due to S. pneumoniae and H. influenzae was found in the 1-3 years group, while the incidence of LRTI due to K. pneumoniae, E. coli, S. aureus and E. cloacae was higher in patients under 1 year of age. Antibiotic susceptibility tests showed that rates of penicillin non-susceptible S. pneumoniae, ampicillin resistant H. influenzae, oxacillin-resistant S. aureus and ESBL-positive K. pneumoniae and E. coli were 55.0%, 16.5%, 41.2%, 42.6% and 4.5%, respectively.

CONCLUSIONSS. pneumoniae, H. influenzae, K. pneumoniae, E. coli and S. aureus were common pathogens of LRTI in children. The infection rate varied with age and gender. Antibiotics for treating LRTI should be selected based on the drug susceptibility test.

Adolescent ; Bacteria ; drug effects ; isolation & purification ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Microbial Sensitivity Tests ; Respiratory Tract Infections ; microbiology ; Seasons

Objective: To analyze the epidemiological characteristics of coronavirus disease 2019 ( COVID-19 ) clusters in Lishui, so as to provide basis for the prevention and control of COVID-19 clusters. Methods: The data of COVID-19 clusters in Lishui from January 23 to March 29, 2020 were collected through China Disease Control and Prevention Information System-Public Health Emergency Information System, and analyzed time, space, scale, source of infection, exposure and transmission route by descriptive epidemiological method. Results: There were 31 cases in 8 clusters ( about 4 cases per cluster ), with no death. The report time was bimodal, peaked first from January 20 to February 10 with 4 clusters imported from domestic and peaked second from March 1 to 29 with 4 clusters imported from overseas. Qingtian County reported 4 clusters, Liandu District, Yunhe County, Qingyuan County and Jingning County each reported 1 cluster. Thirteen cases were restaurant employees, accounting for 41.94%. The cases were mainly occurred in the condition that exposed in the same family ( 6 clusters ), in the same dinner and car ( 1 clusters ), and in the same party ( 1 clusters ). The exposure modes that caused more cases infected were through the same family (9 cases) and through the same dinner and car ( 6 cases ). There were 3 clusters with first-generation cases, 3 clusters with second-generation cases and 2 clusters with third-generation cases. The recurrence rate of the 8 clusters ranged from 1.49% to 7.69%, with a median of 3.47%. Conclusions The COVID-19 clusters in Lishui imported from domestic in the early stage and later from overseas. Most cases were reported from Qingtian County, were engaged in catering business, and exposed by living with families.