Objective To clone and express the hexon protein of three prevalent human adenovi -rus strains causing respiratory disease and analyze the antigenic characteristics of the recombinant proteins . Methods The full length genes encoding hexon protein of human adenovirus serotype 3(HAdV3), serotype 4(HAdV4) and serotype 7(HAdV7) were cloned by PCR and sequenced , respectively.The alignment anal-ysis was performed by using hexon gene sequences from GenBank .The major antigenic regions of hexon pro-tein of the three serotypes were expressed in E.coli and purified.The antigenicity, immunogenicity and cross reactivity of the recombinant proteins were determined by ELISA and Western blot assay .Results The full length gene sequence encoding hexon protein of human adenovirus serotype 4 was firstly reported in China , which showed more than 99%homology in both nucleotide and amino acid sequences with the human adeno-virus type 4 NHRC3 strain.The partial hexon protein sequence of HAdV 3, HAdV4 and HAdV7 containing all of the 7 hyperviriable regions ( HVRs) were expressed in E.coli, respectively .The purified recombinant proteins could be recognized by antiserum of the three serotypes of adenovirus .The antiserum samples against the three recombinant proteins could cross-react with particles of the three serotypes of adenovirus . The possible type-and species-specific epitopes were predicted .Conclusion The major antigenic regions of hexon protein of the three serotypes were successfully expressed .The purified recombinant proteins contai-ning both intertypes and type-specific epitopes showed a strong immunogenicity .

Objective To study the changes of T-C of apolipoprotein CⅢ (ApoCⅢ)-455C gene and to de-tect the relationship between essential hypertension (EH) and the quantity of ApoCⅢ-455C gene. Methods 606 subjects were divided into hypertension group(n=306) and non-hypertension group(n=300). And the two groups 606 were divided into subgroup A (n=90) who had hypertension companied with hypertriglyceridem, subgroup B (n=216)who had hypertension only, subgroup C (n=51) who had hypertriglyceridem only and subgroup D (n= 249) who had none of the two diseases, by the concentration of triglycerides (TG) (TG≥1.69 mmol/L). The quan-tity of ApoCⅢ-455 C was detected by Real Time PCR. TC, TG, HDL-C, LDL-C, ApeA, ApoB 100, ApoCⅢ, ApoE, blood glucose,plasma insulin and waist circumference were measured in all cases. Results There was no difference in the quantity of ApoCⅢ-455C gene between group B and the group D [(45.16±5.97) vs (46.21±6.13)] (P>0.05). The concentration of ApoCⅢ, ApoB100 and the quantity of ApoCⅢ-455C gene in group A were higher than those in group B [0.14±0.03]g/L, (95.00±15.69)g/L, (28.13±4.11) vs (0.09±0.03)g/L,(81.22± 11.87)g/L,(45.16±5.97)(P<0.01)]. It was similar in ApoE [(0.05±0.01) g/L vs (0.04±0.01)g/L] and LDL-C [(2.54±0.61)mmol/L vs (2.15±0.41)mmol/L] (P<0.05). Conclusions The quantity of ApoC Ⅲ-455C gene is not related to EH.

The background,features,dimension and contents of 22 international appraisal tools for clinical guidelines were analyzed,and their history,evolution,rules and problems were described,in order to provide the evidence for developing appraisal tools for domestic clinical guidelines. In the study,it was found that the guideline evaluation tools have been classified and there is a constant strive to achieve the balance between comprehensiveness and practicability. But the existing evaluation tools are insufficient for clinical practical contents of guidelines. The scoring system should be improved. In addition,conflicts of interest,value choice and patients' involvement should be considered increasingly, and should become an important part of the guideline evaluation tools.

[Objective] To determine the sun protection factor (SPF) and protection factor of ultraviolet A (PFA) of a self-made sunscreen cream.[Methods] The sample of a self-made sunscreen cream and a commercial sunscreen cream were painted to an artificial skin (3M film) at a dose of 2 μl/cm2,and a tester was used to evaluate their photoprotective effect.A total of 48 albinism guinea pigs were classified into 8 groups to remain unprotected or be protected by the self-made or commercial sunscreen cream.A solar ultraviolet light simulator (SUV-1000) was used to simulate the ultraviolet rays in sunlight to irradiate the guinea pigs,and the photoprotecfive effect of these sunscreen creams was determined.[Results] The in vitro evaluation revealed that the SPF value of the self-made sun-screen cream and commercial cream was 32.26 ± 2.42 and 30.87 ± 2.57respectively (n =5,t =0.94,P ＞ 0.05),and the PFA value was 24.28 ± 2.44 and 17.53 ± 2.28 respectively (n =5,t =4.52,P＜ 0.01 ).As the in vivo experiment showed,the SPF value of the self-made sun-screen cream and commercial cream was 30.39 ± 6.65 and 28.79 ± 7.36,respectively (n =12,t =0.38,P＞ 0.05),and the PFA value was ＞ 8.91 and ＞ 8.93 respectively.[Conclusion] The photoprotective effect of the self-made sunscreen cream is similar to that of the commercial cream.

Objective To construct a hexon-chimeric human adenovirus type 3 ( HAd3 ) vector expressing two neutralizing epitopes of hepatitis B surface antigen preS 1 (HBsAg-preS1) and to analyze the antigenicity of the chimeric epitopes .Methods Two neutralizing epitopes of HBsAg-preS1 including KR359 and KR127 were inserted into hypervariable region 1 ( HVR1) and hypervariable region 2 ( HVR2) of HAd3 hexon .Chimeric hexon gene encoding the two epitopes was amplified by overlap PCR and then subcloned in -to shuttle plasmid pBR322-L/R containing the homologous recombination regions .The digested shuttle plas-mid containing chimeric hexon gene was co-transfected into Escherichia coli BJ5183 cells together with back-bone plasmid pBRAdΔE3GFP to construct pBRAdΔE3GFP-preS1 vector.Then pBRAdΔE3GFP-preS1 vector was digested with AsiSⅠand transfected into AD293 cells to construct recombinant virus (rAD3E-preS1). CsCl gradient centrifugation was used for purification .Glutathione S-transferase ( GST ) fusion protein KR359KR127 ( GST-KR359KR127 ) was expressed in Escherichia coli BL21 by using expression vector pGEX-4T3.Female BALB/c mice at age 6-8 weeks were intraperitoneally injected with 1010 virus particles or 80 μg GST fusion protein .Serum samples were collected to analyze the antigenicity of two epitopes by ELISA and Western blot .Results ELISA showed that KR 359 and KR127 were successfully exposed on viral sur-faces by using hexon-chimeric HAd3 vector .The induced polyclonal antibodies in serum samples could rec-ognize GST fusion protein and native HBsAg from patients infected with hepatitis B virus .Conclusion The antigen capsid-incorporation strategy could be used to display epitopes on viral surface .Enhanced antigen-specific responses could be achieved through inserting multiple foreign epitopes into hexon HVRs .This study provided evidence for further application of hexon -chimeric human adenovirus type 3 vector in the developmentof vaccine, especially for the development of multivalent hepatitis B vaccine .

Objective To construct a recombinant human adenovirus type 3 ( HAd3 ) vector ex-pressing one major epitope of dengue virus type 1.Methods The gene encoding the envelope protein (304-314 aa) of dengue virus type 1 was inserted into the hypervariable region 1 ( HVR1 ) of HAd3 hexon by using overlap PCR.The recombinant gene was cloned into the shuttle plasmid, then linearized with AsisⅠrestriction enzyme and co-transformed into Escherichia coli BJ5183 strains with the digested backbone plas-mid for homologous recombination.The recombinant plasmid pBRAdΔE3GFP-DENV1 was transfected into AD293 cells to rescue recombinant adenovirus strains (rAdΔE3GFP-DENV1).ELISA and Western blot as-say were performed to evaluate the humoral responses induced in BALB/c mice after the immunization with rAdΔE3GFP-DENV1 strains.Results The recombinant adenovirus strains were successfully rescued. ELISA and Western blot assay showed that the antibodies in serum sample could recognize dengue virus type 1 strains.Conclusion The recombinant adenovirus strains expressing the epitope of dengue virus type 1 were successfully constructed.This study provided evidence for the development of multivalent vaccines against dengue virus.

Since the attenuated live vaccine against measles was developed,the epidemic of measles has been controlled effectively,however,there is a trend of gradual increase of measles cases in recent years.The epidemiological and clinical features of 4430 measles patients in Shenzhen Municipality in last 10 years were reviewed.The data showed that the epidemic season was postponed with the peak of June to September;the prevalent age groups were infants and adults,the number of severe cases increased;and the positive rate of serological antibody in infants with measles was the lowest.

Objective To explore the influence and clinical significance of chemotherapy on the expressions of micro-vessel density (MVD) and vascular endothelial growth factor (VEGF) in pediatric difficult removal solid tumor.Methods From Jan.2002 to Jan.2012,totally 80 cases of the hospitalized pediatric patients with solid tumor (Ⅲ b-Ⅳb stage) were enrolled in this study in Department of Pediatric Surgery of the Second Hospital of Hebei Medical University.All of the tumors were identified as difficult removal malignant tumor by imageology and physical examinations.After biopsy of the tumors,these patients were divided into neuroblastoma group (25 cases),hepatoblastoma group (25cases) and Wilms' tumor group (30 cases).All patients in these 3 groups underwent chemotherapy respectively according to their different pathologic findings,thereafter,the tumor resections were performed.The resection rates of tumor in each groups were recorded and the expressions of MVD and VEGF between preoperative biopsy and postoperative specimens were detected and compared by immunohistochemistry in different groups.Results With chemotherapy,the resection rate of neuroblastoma was 80.00％ (20/25 cases),hepatoblastoma was 92.00％ (23/25 cases) and Wilms' tumor was 96.67％ (29/30 cases).There was no significant difference among them (x2 =4.286,P =0.117).Before chemotherapy,the positive expressions of VEGF in neuroblastoma,hepatoblastoma and Wilms' tumor were 80.00％ (20/25 cases),90.00％ (18/20 cases) and 83.33％ (25/30 cases) respectively,at the same time,the values of MVD were 48.39 ± 15.87,52.16 ± 16.12 and 46.82 ± 19.07.After chemotherapy,the expressions of VEGF and MVD in neuroblastoma,hepatoblastoma and Wilms' tumor were 52.00％ (13/25 cases),45.00％ (9/20 cases),30.00％ (9/30 cases) and 31.31 ± 14.21,25.19 ± 15.31 and 29.06 ± 13.05.Compared with the expressions of them in each group before chemotherapy,significant differences were found (all P ＜ 0.05).Conclusions Chemotherapy can efficiently promote the resection rate for pediatric difficult removal solid tumor.MVD and VEGF can be considered as the markers in evaluating the outcomes of chemotherapy and promoting the resection rate of tumors.

Objective To assess the pituitary function in patients with pituitary adenoma after transsphenoidal ectomy of hypophysoma.Methods Data of 106 patients with pituitary adenoma who were admitted in endocrine department and underwent the operation in PLA General Hospital from January 1993 to January 2010 were collected.Assessments of pituitary function were made before and after surgery.Results Total 23.6％ and 16.0％ of 106 patients underwent pituitary function evaluation by 1 week and 3 months after surgery,respectively.23.5％ and 5.9％ of patients with hyopituitarism before surgery underwent pituitary function evaluation by 1 week and 3 months after surgery respectively,and the respective figures in those without hypopituitarism were 23.6％ and 20.8％.The incidences of new onset of hypopituitarism among 106 patients that underwent surgical procedure were 48.0％ within 1 week after surgery and 35.3％ by 3 months after surgery.Conclusion The rate of re-evaluation of pituitary function by 1 week and 3 months after pituitary surgery was low.Probably,many patients were missed the diagnosis of hypopituitarism after pituitary surgery.

Objective To evaluate the effect of domestic wire-reinforced epidural catheter on the occurrence of adverse events during epidural block.Methods Three hundred ASA Ⅰ or Ⅱ patients of both sexes,aged 18-64 yr,weighing 41-78 kg,scheduled for elective operations under combined spinal-epidural anesthesia,were randomly divided into 3 groups (n =100 each):polyvinyl chloride epidural catheter group (group A),imported wire-reinforced epidural catheter group (group B) and domestic wire-reinforced epidural catheter group (group C).Combined spinal-epidural anesthesia was performed routinely.The corresponding epidural catheter was inserted in each group.The catheterization without difficulty,paresthesia during catheterization,the number of patients in whom blood or cerebrospinal fluid was withdrawn from the epidural catheter,intravascular catheter insertion,injection obstruction,easiness during removal of the catheter,bleeding after removal,postoperative paresthesia and epidural hematoma within 1 week after operation were recorded.Results Compared with A group,the incidences of paresthesia during catheterization,the number of patients in whom blood or cerebrospinal fluid was withdrawn from the epidural catheter,injection obstruction and postoperative paresthesia were significantly decreased (P ＜ 0.05),and no significant change was found in the other parameters in B and C groups (P ＞ 0.05).There was no significant difference in all the parameters between B group and C group (P ＞ 0.05).Conclusion Domestic wire-reinforced epidural catheter can decrease the occurrence of catheterization-induced damage to the nerve and blood vessels and the efficacy is comparable with that of imported wire-reinforced epidural catheter.