Objective To select an optimal non-specific antigen blocking method by using immuno-infiltration assay so as to suit protein chip preparation. Methods Human papillomavirus type 16 L1 protein expressed by insect-baculovirus espressin system was incubated with skimmed milk powder, calf serum, bovine serum albumin (BSA) combinations of five kinds of methods to block the non-specific antigen. PBS was used as control. The effect of eliminating non-specific stain was detected by immuno-infiltration assay. Results After repeated tests, the results showed that the stability and repeatability of blocking effects were poor for the fixing up antigen first and then blocking method, and the blank control was prone to false positive. The infiltration rate of NC membrane would be affected by using skimmed milk powder as a blocking agent because the pore of NC membrane was easily plugged by milk powder particles. The use of calf serum as a blocking agent made it very difficult to determine the result because the calf serum absorbed by NC membrane produced the background; however, when 20g/L BSA was used to blocking before fixing up antibody, the results became satisfactory. Conclusion Fixing up antibody after blocking in immuno-infiltration assay showed that the blocking effect against non-specific antigen was satisfactory, stable and repeatable, indicating this method is a novel optimal blocking method compared with others.

Objective To construct eukaryotic expression vector of HPV18 L1- E6, E7 chimeric gene and examine the humoral and cellular immune responses induced by this DNA vaccines in mice. Methods The C-terminal of major capsid protein L1 gene and mutant zinc finger domains of early E6/7 oncogenes in HPV18 were integrated and inserted into eukaryotic expression vector pVAX1 to generate vaccines pVAX1-L1E6Mxx, E7Mxx. CHO cells were transiently transfected with the individual construct. Target protein expressions in the lysate of the transfected cells were measured by ELISA and immunocytochemistry. After BALB/c mice were vaccinated with various recombinant plasmids(pVAX1-L1-E6M3 or pVAX1-L1-E7M3) and immunie adjuvants (pLXHDmB7-2 or LTB) through different administration routes (intramuscular or intranasal) , the great cellular immune responses were produced as revealed by delayed-type hypersensitivity (DTH) and lymphocyte proliferation, and the expression of IL-4 and IFN- γ cells in CD4+ and CD8+subpopulations. Results The highly efficient expression of pVAX1-L1E6Mxx, E7Mxx vector in host eukaryotic cells were demonstrated both by ELISA and immunocytochemistry. The level of specific serum IgG against HPV in experiment groups mice was much higher than that of control group, and intranuscular immunization group had the highest antibody level. Intramuscular immunization groups were superior to intranasal immunization groups in DTH response, splenocyte proliferation and CD8+ IFN-γ + cells number, but CD4+ IL4+ cell number was higher in intranasal immunization groups. The immunization groups using pLXHDmB7-2 as adjuvant were superior to other groups in immunoresponse. Conclusion These DNA vaccines produce remarkable cellular and humoral immuneresponses in the mouse and may provide as prophylatic and therapeutic candidates for HPV induced cancer treatment.

Objective To explore the effect of the cystoscope indwelling double J tube for the treatment of pregnancy with ureteral calculi,and to observe and analyze the prognosis,and provide the basis for clinical therapy. Methods 46 pregnant women with ureteral calculi were selected.All patients were treated by cystoscope indwelling double J tube.Observation indexes were the follow -up complications after a week of operation time,operation situa-tion,Visual analogue scale (VAS)was used to evaluate the degree of pain operation before and after treatment 48 h. To observe the pregnant women with double J tube placement and delivery status.Results 46 patients were success-fully catheter,the average operation time was (29.14 ±3.27)min.37 patients with lumbar cramps and fever and other symptoms were relieved after treatment.6 patients with replacement of double J tube 2 months later.After treatment, VAS score was (1.16 ±0.83)points,which was lower than (6.93 ±1.02)points before treatment,the difference was statistically significant (t =9.64,P <0.05).After treatment,the infection rates of hydronephrosis and urinary tract were 4.35% (2 /46)and 0% (0 /46),which were lower than before treatment [(15 /46,32.61%)and 19.57%(9 /46)],the differences were statistically significant (χ2 =8.33,9.05,all P <0.05);46 cases of pregnant women were smooth production,no abortion complications.The average indwelling double J tube time was (5.08 ± 1.16)months.Conclusion Cystoscope indwelling double J tube for the treatment of ureteral calculi in pregnancy has less invasion,high stone clearance rate,less postoperative complications and high safety,the prognosis was good,and it is a safe and effective method.

A rare case of adult granulosa cell tumors of testis with metastasis of pulmonary and retroperitoneal lymph nodes was reported. The patient was 26 years old, who underwent resection of the affected testis by orchiectomy and postoperative radiotherapy, and pulmonary and retroperitoneal lymph node metastases were detected 5 years after surgery by regular follow-up. Adult granulosa cell tumors of testis has the potential of malignancy, and the prognosis is poor if the distant metastasis occurs.

Objective:To observe the clinical effect of Nie-pinching spine coupled with point application and injection for cerebral palsy (CP) in children with spleen deficiency.
Methods:A total of 70 CP children with spleen deficiency were randomly allocated into a treatment group (n=35) and a control group (n=35). Children in the treatment group were treated with comprehensive rehabilitation training, Nie-pinching spine, point application and point injection, whereas children in the control group were treated with comprehensive rehabilitation training and oral administration of multi-enzyme tablets. One course of treatment made up of 20 d. There was a 20-day interval between two courses. The cases were treated for a total of 3 courses. Then the therapeutic efficacies were evaluated using scores of spleen-deficiency symptoms and gross motor function measure (GMFM).
Results:After 3 courses of treatment, the total effective rates for spleen-deficiency symptoms and rehabilitation were 85.7% and 77.1% respectively in the treatment group, versus 51.4% and 57.1% respectively in the control group, showing statistical differences (bothP<0.05).
Conclusion:In addition to comprehensive rehabilitation training, Nie-pinching spine combined with point application and injection obtained better effects than oral administration of multi-enzyme tablets.

Objective To identify distinct subgroups of military personnel from Paracel Islands based on mental status for providing basis to intervention measures.Methods 174 enlisted military personnel were interviewed by symptom checklist-90 (SCL-90).The factor scores of SCL-90 were compared with army man norm and control group,then the cluster analysis was conducted.Results Interpersonal sensitivity,phobic and psychoticism of military personnel on Paracel Islands((1.65±0.56),(1.24±0.33),(1.44±0.46)) scored lower than those of army man norm (P＜0.05).Interpersonal sensitivity of military personnel on Paracel Islands scored lower than those of control (P＜0.05),depression and anxiety of them had no difference to control (P＞0.05),while other factors of them scored higher than control.According to cluster analysis,174 military personnel were divided into three subgroups.The first subgroup with high scores for all nine SCL-90-R dimensions,the second cluster showed moderate scores and the third cluster had lowest scores.Statistically significant sociodemographic differences could be found between the cluster groups (P＜0.05).Conclusion The mental health status of military personnel stationed in Paracel Islands is good on the whole,and can be divided into three clusters with different demographic and service characteristics.

Objective Attenuated strains of Shigella are attractive live vaccine candidates for eliciting mucosal immune responses which is a suitable carrier for the prophylactic human papillomaviruses (HPV) vaccine development, To examine the potential of a live Shigella based prophylactic HPV vaccine, HPV16L1should be expressed in attenuated shigella strain. Methods A Shigella large invasive plasmid (icsA/virG) based prokaryotic expression plasmid pHS3199 was constructed. HPV16L1 gene was inserted into plasmid pHS3199 to form pHS3199-HPV16 L1 construct, and pHS3199-hpv16L1 was electroporated into a live attenuated shigella strain sh42. The expression of HPV16L1 protein was demonstrated by Western blotting with monoclonal antibody to HPV16L1, The genetic stability of recombinant strain sh42-HPV16 L1 was monitored by consecutive passage culture. Invasive ability of sh42-HPV16L1 was evaluated by Hela cell infection assay. Results HPV16 L1 protein can be expressed in recombinant strain sh42-HPV16 L1, and the protein stably expressed over 140 generations. The invasive ability of sh42-HPV16L1 was diminished dramatically compared to its parent strain, but not abolished completely. Conclusion HPV16L1 protein was constitutively expressed in the attenuated strain of shigella flexneri sh42, and maintained partial invasive ability. Our strategy may represent a promising vaccine candidate against genital HPV16 infection.

Objective: To observe the clinical effect of acupuncture plus occupation therapy for fine motor functions in children with spastic cerebral palsy. Methods:A total of 80 cerebral palsy kids with fine motor dysfunction were allocated into two groups by envelop, 40 cases in each group. Cases in the control group were treated with occupation therapy. Based on the therapy given to the control group, cases in the treatment group were supplemented with acupuncture at the points of the three yang meridians of hand. The treatment was done every other day and 10 times for a course. There was a 15-day interval between two courses, and 3 courses in total. The therapeutic efficacies were evaluated using Peabody developmental motor scale 2 (PDMS-2) and modified Ashworth scale (MAS) before and after treatment. Results:After treatment, the standard score for grasping (Grs), standard score for visual-motor integration (Vis), fine motor quotient (FMQ) and modified Ashworth scale (MAS) were significantly improved in both groups (P<0.05). The improvement in the treatment group was more obvious than that in the control group (P<0.01). Conclusion:Acupuncture plus occupation therapy can achieve better effect than occupation therapy alone in improving fine motor functions of upper limbs in cerebral palsy kids.

Objective To isolate bacteriophages against extensively-drug resistant Acinetobacter baumannii from hospital sewage and analyze their biological characteristics.Methods Extensively-drug resistant Acinetobacter baumannii isolated from several hospitals in Shanghai during December, 2013 to July, 2014 were used as host bacteria, adopting double-layer agar method to isolate bacteriophages from raw sewage of these hospitals.The phage with broad host range was selected for further study, including observation of electron microscopic morphology, examination of thermal stability, pH stability and the optimal MOI, drawing of the adsorption, one-step-growth and infection curves, as well as sequencing of the phage genome DNA. Results An extensively-drug resistant Acinetobacter baumannii bacteriophage vB_AbaP_PD-AB9 ( PD-AB9 for short) with broad host range was isolated, and electron microscopy revealed it belonged to Podoviridae family.The optimal MOI of PD-AB9 was 0.001.PD-AB9 remained stable among 4 ℃to 50 ℃and pH 4 to 11.In the adsorption experiment, the adsorption rate of PD-AB9 reached above 95%within 5 min.PD-AB9 had a latent period of 4 min and a burst size of 213.PD-AB9 could obviously restrain the host growth, with faster effect at the higher MOIs (MOI=1, 0.1, 0.01) than at the lower ones (MOI=0.001, 0.000 1).Furthermore, genome of PD-AB9 proved to be a double-stranded linear DNA with size of 40 938 bp and GC content of 39.34%.Conclusions PD-AB9 exhibits good thermal stability, wide pH tolerance range, very fast adsorption, a short latent period, a large burst size and it could quickly cause effective host lysis after infection.Therefore, PD-AB9 is promised to act as a new antimicrobial agent to control drug-resistant Acinetobacter baumannii infections and its bio information remains to be further studied.

Anastomosing hemangioma is a newly reported variant subtype of capillary hemangioma, and its biological behavior is mostly benign. We reported a case of anastomosing hemangioma originating from the kidney. After nephrectomy, the patient was followed up for 13 months, and no recurrence or distant metastasis was observed.