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.

Low back pain (LBP) is one of the most common types of disability affecting individuals in Western countries, and the as-sessment of LBP-related disabilities represents a significant challenge. The disability assessment scales in LBP are commonly used in clini-cal diagnosis and treatment as well as evaluation system for clinical research, such as Oswestry Disability Index (ODI), Roland Morris Dis-ability Questionnaire (RMDQ), and Quebec Back Pain Disability Scale (QBPDS). This article focused on the clinical applications, reliability and validity, strength and limitations of these scales, to help the clinical application and scientific research for the LBP disability scale.

Objective To compare volumetric size, conformity index (CI), degree of inclusion (DI) of internal gross target volumes (IGTV) delineated on 4D-CT-MIP and PET-CT images for primary thoracic esophageal cancer. Methods Fifteen patients with thoracic esophageal cancer sequentially underwent enhanced 3D-CT, 4D-CT and PET-CT simulation scans. IGTVMIP was obtained by contouring on 4D-CT maximum intensity projection ( MIP). The PET contours were determined with nine different threshold methods (SUV≥2?0, 2?5, 3?0, 3?5), the percentages of the SUVmax(≥20%, 25%, 30%, 35%, 40%) and manual contours. The differences in size, conformity index (CI), degree of inclusion ( DI) of different volumes were compared. Results The volume ratios ( VRs) of IGTVPET2. 5 to IGTVMIP , IGTVPET20% to IGTVMIP, IGTVPETMAN to IGTVMIP were 0?86, 0?88, 1?06, respectively, which approached closest to 1. The CIs of IGTVPET2?0,IGTVPET2.5,IGTVPET20%,IGTVPETMAN and IGTVMIP which were 0?55, 0?56, 0?56, 0?54,0?55, respectively, were significantly larger than other CIs of IGTVPET and IGTVMIP (Z= -3?408-2?215,P <0?05). There were no statistical significance in the DIs of IGTVMIP and IGTVPET2.5,IGTVMIP and IGTVPET20%, IGTVMIP and IGTVPETMAN(0?77,0?82,0?71,0?67, 0?68,0?82,P>0?05). Conclusions The targets delineated based on SUV threshold setting of≥2?5, 20% of the SUVmax and manual contours on PET images correspond better with the target delineated on maximum intensity projection of 4D-CT images than other SUV thresholding methods.

Objective To study bacterial conditions in amniotic cavity of the third trimester and the delivery. Methods Patients underwent cesarean section due to various reasons were randomly enrolled in the study. Ten ml aqua amnii taken from using aspesis were cultured and the bacteria were identified and tested for drug sensitivity. All patients were not treated with antibiotics before and during operation,conventional antibiotics treatnent was used after operation. The patients were divided into 3 groups based on their status: 34 cases of premature rupture of membrane ( PROM ), thirty-four cases of labor without PROM, and 27 cases of pregnancy without PROM. The positive rate, species , distribution and drug sensitivity of bacteria in the 3 groups were analyzed. Results The positive rate was 61.8% (21/34) in PROM ,52. 9% ( 18/34 ) in labor without PROM,11.1% (3/27)in pregnancy without PROM. The positive rates were significantly different among the 3 groups (x2 = 17.29 ,P =0.000). However, there was no significant difference between the labor without PROM group and the pregnancy without PROM group ( x2 = 0.541 , P = 0.462 ). Staphylococcus epidermidis was the most common isolated bacteria, following by staphylococcus aureas, streptococcus, colon bacillus, and pseudomonas. There were no significant differences of bacterial species between labor without PROM and pregnancy without PROM( x2 = 11.9,P =0.535 ). The relative higher positive rate in the labor without PROM and PROM indicated that they were important inducement of bacteria infection. There were no significant difference on positive rate and bacteria species distribution between the PROM and labor without PROM group,which suggested that up-bound along vagina to amniotic cavity was the most common route of infection. Sensitive antibiotics should be given to the patients of PROM and labor without PROM targeted at staphylococcus,streptococcus, colon bacillus and monad. Antibiotics such as Cephems, Penicillins and Clindamycin were sensitive to various bacteria and safe to both mother and baby, thus were recommended. Quinupristin, Vancomycin,Furantoin, Fusidic ,Teicoplanin, Amikacin and Meropenem were also sensitive to various bacteria, but due to their side effects and the principle of ladder antibiotics using, they were recommended as second line antibiotics. Conclutions Up-bound along vagina to amniotic cavity is the most common route of infection. For rupture of membrane and labor without PROM patients, antibiotics should be given, and strict aseptic technique,washing cavity and incision, using antibiotics after operation are necessary to prevent infection.

AIM:To explore the protective effect of ischemic preconditioning on myocardial capillary bed. METHODS: Anesthetized open-chest dogs in ischemic preconditioning (IP) group ( n =6) were subjected to 5 min ischemia and 5 min reperfusion for 4 times in left descending coronary artery (LDA) while the dogs in sham-operated (SO) group ( n =6) were observed for 40 min without any stimulating. Myocardial contrast echocardiography (MCE) was performed before the surgery, at the end of the first and the fourth ischemia and reperfusion period, respectively. Myocardial samples in ischemic area were obtained for capillaries ultrastructure examination and their quantitative analysis was conducted by corresponding imagine analyzing system, then compared within group and between control group. RESULTS: (1) Comparing fourth 5 min ischemia with first one, the percents of area defect in minimum (AD min%) by MCE decreased from 32.6%?5.7% to 21.8%?5.2%( P

Objective To investigate the role of heparin binding epidermal growth factor-like growth factor(HB-EGF)in active psoriasis vulgaris.Methods HB-EGF mRNA and protein were detected by hy-bridization in situ and immunohistochemistry in normal skin tissues,lesional and non-lesional psoriatic skin of progressive stage.Results In normal skin tissues,the stain of HB-EGF mRNA and protein was located in the basal layer of epidermis(100.00%)and there was only a little expression in the suprabasal layers(16.67%).Focal overexpression was found in the suprabasal layers of non-lesional and peri-lesional psoriat-ic skin(88.00%,80.00%respectively);however,there was no HB-EGF mRNA protein expression in the superabasal layers of the central part of psoriatic lesions(0),and nearly no expression in the basal layer(4.00%).Conclusion HB-EGF may play an important role in the pathogenesis of early psoriasis.

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 investigate the infection status and genotype distribution of cervical human papillomavirus (HPV) in women of different ethnic groups and different ages in Yili, Xinjiang Uygur Autonomous Region (Xinjiang). Methods: By using the convenient sampling method, 54 760 women from November 2015 to May 2017 seeking for service in gynecological clinics in a general hospital in Yili, Xinjiang, were selected as the research subjects, and 3 445 samples of cervical mucous exfoliative cells were collected, and the social information of their ethnic and age was collected at the same time. The inclusion criteria were those with sexual life, cervical integrity, and ethnic groups for Han or Uygur or Kazak. PCR-reverse dot blot hybridization was used to detect HPV genotyping in exfoliated cells, and chi-square test was used to compare the difference of HPV positive rate among different ethnic groups. Then, according to ethnicity and age, the differences in positive rates of different ages and ethnic groups were compared in each layer. Results: The positive rate of HPV was 25.6% (882 cases), of which the Han, Uygur and Kazakh were 27.9% (564 cases), 22.9% (196 cases) and 21.6% (122 cases), and the difference was statistically significant (χ²=13.80, P=0.001). The most prevalent high-risk genotypes of Han women were HPV16/52/58, accounting for 24.8% (140 cases), 17.7% (100 cases) and 9.8% (55 cases), respectively. The most prevalent high-risk genotypes of Uygur women were HPV16/52/53, accounting for 34.2% (67 cases), 12.8% (25 cases), 9.2% (18 cases), respectively. The most prevalent high-risk genotypes of Kazak were HPV16/52/53, accounting for 37.7% (46 cases), 17.2% (21 cases), 12.3% (15 cases), respectively. The highest rate of HPV in Uygur patients aged ≥61 years was 41.5% (22 cases), and the lowest in group 36-40 years old, 15.9% (21 cases), the difference between different age groups was statistically significant (χ²=35.01, P<0.001). Conclusion The positive rate of HPV infection among Han, Uygur and Kazak in Yili Prefecture of Xinjiang was different, and the HPV positive genotype differs among different ethnic groups.

OBJECTIVETo investigate which of the two immunoglobulin (Ig)-like domains, the immunoglobulin variable region homologous domain IgV (hB7.2 IgV) and the immunoglobulin constant region homologous domain IgC (hB7.2 IgC) on the human B7.2 molecule contains receptor binding sites, and to evaluate whether the B7.2 protein expressed in bacteria has biological activity in vitro.

METHODSThree fragments of hB7.2 IgV,hB7.2 IgC and the complete extracellular region of human B7.2 containing both the IgV and IgC domains,hB7.2 Ig (V+C), were amplified by PCR and subcloned into pGEM-Teasy. Three recombinants,pGEX-4T-3-hB7.2 IgV,pGEX-4T-3-hB7.2 IgC and pGEX-4T-3-hB7.2 Ig (V+C), were generated by cloning the fragments into a prokaryote expression plasmid (pGEX-4T-3) and transformed into the host strain E. coli DH5alpha. The relevant target fusion proteins consisting of GST and hB7.2 IgV,hB7.2 IgC and hB7.2 Ig (V+C), were identified by SDS-PAGE and Western blotting. With the presence of the first signal imitated by anti-CD3 antibody, T cell activation was observed by exposing purified T lymphocytes to each soluble form of the three bacterially-produced human B7.2 fusion proteins by [(3)H]-TdR incorporation.

RESULTSThree recombinant fusion proteins of human B7.2, GST-hB7.2 IgV, GST-hB7.2 IgC and GST-hB7.2 Ig (V+C) were produced and detected in inclusion body form from engineered bacteria. With the first signal present,T lymphocytes proliferated when co-stimulated by bacterially-produced either GST-hB7.2 Ig (V+C) or GST-hB7.2 IgV fusion proteins, but not by GST-hB7.2 IgC.

CONCLUSIONSFunctional human B7.2 fusion protein can be produced in bacteria. The IgV-like domain of human B7.2 is sufficient for B7.2 to interact with its counter-receptors and co-stimulate T lymphocytes.

Antigens, CD ; pharmacology ; B7-2 Antigen ; Escherichia coli ; genetics ; Humans ; Immunoglobulin Constant Regions ; pharmacology ; Immunoglobulin Variable Region ; pharmacology ; Lymphocyte Activation ; Membrane Glycoproteins ; pharmacology ; Plasmids ; Recombinant Fusion Proteins ; pharmacology ; T-Lymphocytes ; immunology

Objective To evaluate the function of auditory nerve and brainstem in children with Bell's palsy by using brainstem auditory evoked potential ( BAEP) . Methods A retrospective study was car-ried in total of 69 patients with Bell's palsy admitted in the pediatric department of Shengjing Hospital of China Medical University from January 2017 to January 2018. There were 33 males and 36 females,aged 1 to 12 years old. Sixty healthy children served as control group. BAEP were determined in 69 patients and 60 healthy subjects. EEG and CT or MRI of brain were performed to exclud tumor,intracranial infection and so on. The patients in the case group were divided into 2 groups:1-3 years old group (38 cases) and 4-12 years old group (31 cases). Sixty healthy children were also divided into 2 groups:1-3 years old group (30 cases) and 4-12 years old group (30 cases). Results The hearing threshold was normal in the children of the control group. There was no significant difference not only in potential latency but also internal potential latency of BAEP between right and left ear. The abnormal rate of BAEP was 39. 1％ ( 27/69 ) in case groups,19 cases of them recovered(70. 4％,19/27) after 2 weeks of clinical treatment. Another 8 patients re-covered after 2 months. The abnormality rate of EEG was 5. 8％ (4/69),and the abnormality rate of CT or MRI in head was 14. 5％ (10/69). Both were nonspecific changes. In the two groups,Ⅰ-Ⅲinternal potential latency was significantly shortened ( P <0. 05 ) , and Ⅲ-Ⅴinternal potential latency was significantly pro-longed (P<0. 05)respectively,between the normal side and abnormal side. There were significant differences in the Ⅲ potential latency(P<0. 05),Ⅰ-Ⅲinternal potential latency (P<0. 01),and Ⅲ-Ⅴ internal poten-tial latency ( P <0. 01 ) respectively, between pretherapy and post-treatment. Conclusion Bell's palsy of children could cause abnormal of auditory nerve conduction,and BAEP could find the electro physiologic ab-normality in nervous system,and guide clinical intervention.