Objective:To purify apolipoprotein H (apoH) from fresh human serum with heparin affinity column self-prepared and to prepare polyclonal antibody anti-ApoH.Methods:Affinity column was made by reaction of indirect reductive amination and operating conditions were optimized and their effects were investigated.The apolipoprotein H antigen was purified from human serum by methods of perchloric acid precipitation,ion exchange and affinity chromatography.The polyclonal antiserum against the antigen was obtained by immuning rabbits.Results:The amount of heparin immobilized on the synthesized gel was 5 mg/ml (medium).Molecular weight of the purified antigen was about 50 kD by SDS-PAGE and no other bands were seen,the percentages of 16 amino acids were consistent with that reported previously.In ELISA and dot-ELISA,the polyclonal antibody displayed a cross reaction with bovine serum albumin,while no such reaction was observed in result of Western blot under reduced and denatured condition.Conclusion:The self-prepared affinity column costs very low,but has excellent affinity performance and stable heparin linking,and high purified apoH can be obtained by using the reagent.Polyclonal antibody needs to be further purified before detecting serum apolipoprotein H.

Objective To observe the effect of Sishen Pill on ICAM-1 mRNA and protein expression of colonic mucosa in rats with ulcerative colitis (UC), and identify its mechanism. Methods Taolly 40 SPF Wistar rats were randomly divided into blank group, model group, Pill group and SASP group. Except the blank group, UC model was prepared with TNBS/ethanol enema. Pill group was given Sishen Pill 5 g/kg, and SASP group was given SASP 0.3 g/kg by gavage, blank group and model group was given the same volume physiological saline for three weeks. Morphological injury of colonic mucosa was observed and scored. ICAM-1 gene and protein expression were detected by RT-PCR and immunohistochemical method. Results Inflammation and ulceration were found on the colonic mucous membrane of rats in the model group. The expression of ICAM-1 gene and protein of colonic tissues of rats in the model group increased compared with that of the blank group (P<0.01). Compared with the model group, the expression of ICAM-1 in Pill group decreased (P<0.05, P<0.01). Conclusion Sishen Pill can decrease the expression of ICAM-1 mRNA and ICAM-1 protein, inhibit the infiltration of inflammation cells, prevent and reduce colon tissue damage, and play a vital role in the treatment of UC.

Ubiquitous chromatin opening element ( UCOE ) , composed of the promoters of human housekeeping genes, prevents transgene from silencing and produces consistent, stable and high-level gene expression irrespec-tive of the chromosomal integration site. The research studied the influence of different UCOE element parts on antibody expression in CHO cells. UCOE 1. 5 kb from chromobox homolog 3 ( CBX3 ) , UCOE 2. 5 kb from the heterogeneous ribonucleoprotein A2/B1 ( HNRPA2 B1 ) and the whole UCOE 4. 0 kb were inserted into the anti-body light and heavy chain vectors, respectively, and transfected into CHO cells using antibiotics-Zeocin and Blasticidin for pressure selection. Four groups of monoclonal cells were harvested and antibody expression of each group was detected. The monoclonal cells with UCOE 1. 5 kb and UCOE 2. 5 kb increased 1. 5 to 2-fold in the level of antibody expression, wheareas, monoclonal cells with UCOE 4. 0 kb increased 3 to 4-fold. The enhance-ment of two housekeeping promoter genes on antibody expression could stack up.

Objective To explore the clinical methodology and effects of correction of secondary unilateral cleft lip nasaldeformity by anatomical restoration and cartilage of nasal septum.Methods With the Goodman's incision in the columella and nasal vestibule,we cut out the cartilage of nasal septum and built a new support structure of tip to reshape the nasal shape after the adequate anatomical restoration under the magnifier of 2.5 times.Results The operation corrected alanasi collapse skewed columella and nostrils symmetry,etc.21 cases were treated and followed up for a month and fifteen months,showing stablized effect with columella centerno alanasi collapse,symmetric nostril and no recurrence.Conclusions On the base of nasal anatomy,a good therapeutic effect has been archeived by the adequate anatomical restoration,cutting out the cartilage of nasal septum and builting a new support structure of tip to reshape the nasal shape.

Objective: To analyze the current situation and effect of vertical integrated of rural health services in Dafeng County. Method: Policy documents and data are collected and interviews are implemented for studying the integrated situation. Results: The vertical integration in Dafeng County is virtual joint form with forming 6 city-level medical volunteer service teams and 52 health management teams. With the imple-mentation of fully integrated management, signing service with rural doctor mode is carried out, which pro-mote the establishment of rural unified service standards and regulations. Using health card of residents which promote the regional shared of treatment information. Conclusions: Vertically integrated mode had connection in health management teams, supported by fully integration, developed by signing service with rural doctor and carried by residents’ health card, which could provide reference for implementing vertical integration of rural health service. But strategies needed to be strengthened in improving health services’ ability of township hospitals, clarifying interest relationship as well as integrating health service regulations and strengthen the health information system for sharing and docking.

Objective To explore the diagnostic value of CT cerebral perfusion imaging on the brain injury of the high +Gx in Rhesus. Methods Seven healthy male adult Rhesus were randomly divided into control group and +15 Gx group. The +15 Gx group underwent parabolic G curve in animal centrifuge. The animals were all examined by CT cerebral perfusion before +Gx exposure, 2 h, 24 h, and 1 week after ~+Gx exposure. The results were compared with pathologic examination. Results 2 h and 24 h after +15 Gx exposure, brain ischemia was showed on CT cerebral perfusion imaging. After 1 week, the brain ischemia was almost recovered to normal. Mild ischemic atrophy was observed in pyramidal neurons in cerebral cortex by light microscopy. Electron microscopic observation showed chromatin marginating and mitochondria cristae blurring in pyramidal cells after +Gx overload. Conclusion High G from simulating spaceship emergency return can cause ischemic injuries of the brain in Rhesus, and CT brain perfusion imaging can provide valuable diagnostic information.

In order to verify whether p-nitrophenylalanine-containing BAFF vaccine can be used as a candidate molecule for the treatment of autoimmune diseases with BAFF over-expression,a soluble mutant of B cell activating factor belonging to the TNF Family (smBAFF) and its pNO2Phe mutant(pNO2Phe65smBAFF),which site specific incorporated pNO2Phe at position 65 of smBAFF,were expressed and purified.In order to evaluate the feasibility of using pNO2Phe65 smBAFF to treat BAFF-over-expressed autoimmune diseases,we investigate its Lymphocyte-stimulating capacity,immunogenicity and inhibitory effect of serum on biological activity of natural BAFF.The pharmacological activity of pNO2Phe65 smBAFF was evaluated using a cGVHD(graft-versus-host disease) induced SLE mouse model.Results indicated that pNO2Phe65 smBAFF,could bind to mouse lymphocytes but could not promote the proliferation of mouse lymphocytes.Moreover,the incorporation of pNO2Phe significantly increased the immunogenicity and induced cross-antibody,which can inhibit the biological activity of natural BAFF.In cGVHD induced SLE mouse model,pNO2Phe65 smBAFF can significantly reduce the symptoms of the disease and play a therapeutic role.Therefore,pNO2Phe65 smBAFF can be used as a candidate molecule for the treatment of autoimmune diseases with BAFF over-expression.

B7-H3 is an immune checkpoint molecule overexpressed on the surface of a variety of tumors, and is is an ideal target for tumor immunotherapy. In this study, nitrolated T cell epitope designed in the early stage of the laboratory was used to construct an epitope vaccine that can target immune checkpoint B7-H3. The vaccine can significantly inhibit tumor growth in the CT26 colon cancer model, and has a significant synergistic effect with the PD-L1 protein vaccine. B7-H3 vaccine can increase the proportion of CD4+ T cells in splenic T lymphocytes and the proportion of CD8+ T cells in tumor-infiltrating T lymphocytes, while reducing the proportion of suppressor Treg cells in tumor-infiltrating CD4+ T lymphocytes, which effectively improves tumor immunosuppressive microenvironment. Research results suggest that the B7-H3 epitope vaccine can be used as an effective tumor vaccine candidate molecule.

Objective: To clarify that protection of Tongxinluo against myocardial ischemia/reperfusion injury relates to enhancing endothelial barrier in diabetic rats.
Methods: A total of 32 Zucker diabetic rats were randomized into 4 groups:Sham group, Model group, Insulin group and Tongxinluo group, n=8 in each group. In addition, there was a Control group containing 8 non-diabetic Zucker rats. Myocardial ischemia/reperfusion injury model was established by a 45-min ischemia and 3-h reperfusion protocol. The size of infarction was detected by pathological staining, the microvascular permeability was examined by Miles assay to obtain the lfuorescein isothiocyanate concentration in myocardial tissue, the Triton X-100 soluble and insoluble VE-cadherin was measured by membrane protein extraction and western blot analysis.
Results:The size of infarction in Model group was obviously higher than that in Control group (55.2 ± 1.4)%vs (36.2± 1.3)%,P<0.05 and the lfuorescein isothiocyanate concentration was also higher in Model group than Control group. Both insulin group and Tongxinluo group had the similar size of infarction with Model group (36.8 ± 1.2)%vs (38.7 ± 1.1)%, P>0.05. Both insulin group and Tongxinluo group got lower lfuorescein isothiocyanate concentration (all P<0.01) and VE-cadherin internalization (all P<0.05) than that in Model group.
Conclusion: Protection of Tongxinluo against myocardial ischemia/reperfusion injury in diabetic rats is as effective as
insulin, but the effect is independent of reducing blood glucose and may be related to enhancing endothelial barrier.

Objective To explore the efficacy and safety of 980 nm diode laser therapy for treatment of benign prostatic hyperplasia (BPH). Methods Data of 170 patients with BPH treated with 980nm diode laser system were reviewed. The mean operative time, blood loss, surgical complications, the international prostate symptom score (IPSS), bladder residual urine volume and flow rate changes were collected and analyzed. Results One hundred and seventy cases were safe during the perioperative period. The average operation time was (74 ± 11) min, surgical removal of prostate tissue mass of (54±12) g, blood loss (72±11) ml. There was no TUR syndrome occurred. 170 patients were followed up 2 to 24 months. The IPSS decreased from preoperative 25.0±5.5 to 9.0±2.5. The maximum flow rate increased from preoperative (6.2±2.3)ml/s to post-operative (17.4±3.5) ml/s. The residual urine volume decreased from preoperative (210.0±25.6) ml to postoperative (25.2±4.6) ml. All the differences were statistically significant (P＜0.05). Conclusion Transurethral vaporization of 980 nm diode laser could be a safe and effective treatment modality for BPH.