To investigate the academic features of Waizheng-YiAn-Huibian and provide references for research and application of it. With research approach in bibliography, the academic features of the book were studied from source of medical cases, writing characteristic, academic thought, etc. The academic features of Waizheng-YiAn-Huibian were summarized into the following 5 points, including multiple chosen medical cases but all with unique individual features, precious discussion after each case, named external symptoms, but covered internal symptoms, encouraging mild treatment and against being broad and complicated, emphasis on veins and arteries, positioning and nature determination. Waizheng-YiAn-Huibian is a wonderful masterpiece in medical cases research and worth learning and studying.

To discuss the academic features of Ding-Ganren’s medical records and to provide the references for learning and applying the theories in the book. With bibliography research method, the academic features of Ding-Ganren’s medical records were investigated in academic thoughts, medication characteristic and clinic experience, etc. The academic features of Ding-Ganren’s medical records were summarized into 4 pointes, which are ①good at classical prescription and determination of treatment based in pathogenesis obtained through differentiation of symptoms and signs of typhus and complicated diseases. ②Focusing on six meridians and proposing the combination of cold and warm treatment. ③Mild treatment and claiming gentle application of the medicine. ④Determination of treatment based in pathogenesis obtained through differentiation of symptoms and signs of scarlet fever and emphasizing to distinguish Qi from Ying. Ding-Ganren YiAn has high academic value in research and clinic application and worth further study.

Objective To investigate the factors related to negative conversion of HBsAg in patients with acute hepatitis B (AHB).Meth-ods A total of 106 AHB patients who were admitted to the Department of Infectious Diseases,Zhoukou Central Hospital from February 2007 to February 2013 were recruited.Liver function,five serological markers of HBV infection (HBsAg,HBeAg,anti-HBs,anti-HBe,and anti-HBc),and HBV-DNA were measured every three months.All patients were followed up for 12 months.The major HBV genotypes (A through D)were determined by type-specific primer nested PCR.Categorical data were expressed as rates and compared with χ2 test;continuous data were expressed as mean ±SD and analyzed by t-test.Results When just admitted to our hospital,compared with the pa-tients with negative conversion of HBsAg,AHB patients with HBsAg persisting for more than 6 or 12 months had a significantly lower peak level of alanine aminotransferase (ALT)(718 ±696 vs 1282 ±913 U/L,622 ±514 vs 1203 ±924 U/L,P<0.05)and a significantly high-er level of HBV-DNA (6.8 ±1.4 vs 5.2 ±1.5 log10 copies/ml,7.3 ±1.6 vs 5.4 ±1.5 log10 copies/ml,P<0.05).The HBsAg clear-ance rate in patients with HBV genotype A was significantly lower than that in patients with HBV genotype B or D (75.0% vs 89.5% or 100%,83.3% vs 97.4% or 100%,P<0.05).Conclusion The HBsAg clearance rate in AHB patients may be associated with HBV genotypes,peak level of ALT,and HBV-DNA.

Under the current system, the number of sole foreign -investment hospitals and foreign -domestic joint venture hospitals is growing.They are different from public hospitals in terms of their foperation , market share, medical-service pricing poli-cy and their quality and management of medical care.Mechanisms still need to be developed so that these hospitals could move forward to improve our health-system overall.In the meanwhile, public hospitals should be powered by the patient-centeredphilosophy and improve their management and services to enhance their competiveness.

Objective To study the X-ray and CT representations of primary malignant lymphoma of bone.Methods X-ray and CT representations of primary malignant lympoma of bone in 27 cases proved by operation and pathology,combined with clinic manifestations,operation and pathology were analysed.Results The tumors located in vertebra were 14 cases(51.9%),in tubular bone were 9 cases(33.3%).The imaging findings included bone destroy in 85.7%,bone sclerosis in 7%;only 4 cases appeared light periosteous reaction;one case appeared multiple focuses.Conclusion When the dissolved bone destruction without periosteous reaction,presented soft tissue mass in vertebras or tubular bone are showed on X-ray films or CT,the possibility of bone lymphoma shonld be considered,but this neoplasm is lack of characteristic imaging features.

Objective: To investigate the expression of long non-coding RNA (lncRNA) DLGAP1-AS2 in gastric cancer and its clinical significance. Methods: The data set of gastric adenocarcinoma was downloaded from The Cancer Genome Atlas (TCGA) database, and the difference of DLGAP1-AS2 expression between tumor tissues and normal tissues was analyzed. The tumor and adjacent tissues were collected from 85 gastric adenocarcinoma patients, and the plasma samples were collected from 25 gastric cancer patients and 25 healthy controls. Then the expression of DLGAP1-AS2 in tumor tissues and plasma samples was detected by real-time fluorescent quantitative PCR, and the correlation of the level of DLGAP1-AS2 with the clinicopathological features of gastric cancer patients was analyzed. Besides, after the transfection with DLGAP1-AS2 siRNA, the proliferation, migration and invasion of gastric cancer AGS cells were observed by viable cell counting method, colony formation assay and Transwell chamber assay, respectively. Moreover, the expressions of epithelial-mesenchymal transition (EMT)-related molecules after silencing DLGAP1-AS2 in AGS cells were detected by Western blotting. Results: The expression of DLGAP1-AS2 in gastric cancer tissues was higher than that in normal gastric epithelium according TCGA database (P < 0.001). DLGAP1-AS2 was significantly overexpressed in gastric cancer tissues as compared with the corresponding paracancerous tissues, which was correlated with the advanced TNM stage, lymph node metastasis and overall survival rate (all P < 0.05). While the level of DLGAP1-AS2 was also upregulated in the plasma of patients with gastric cancer as compared with the healthy (P < 0.05). Knockdown of DLGAP1-AS2 expression inhibited the proliferation, colony formation, migration and invasion of gastric cancer AGS cells (all P < 0.05). In addition, the expression levels of matrix metalloproteinase 2 (MMP2), Slug, N-cadherin (N-cad) and Twist were significantly down-regulated in AGS cells transfected with DLGAP1-AS2 siRNA (all P < 0.05). Conclusion: The expression of DLGAP1-AS2 is significantly increased in tumor tissues and plasma of gastric cancer patients. DLGAP1-AS2 knockdown can reduce the proliferation, migration and invasion of gastric cancer cells, and its action mechanism may be related to the regulation of EMT-related genes expression.

Objective To construct the eukaryotic expression plasmid for the expression of human Connexin26 in COS-7 cells.Methods Total RNA was isolated from human peripheral blood lymphocytes and used as template for the PCR cloning of the human Connexin26 gene.The human Cx26 cDNA containing the 678 bp whole coding region of the human Connexin26 gene was amplified by PCR using specific primers and cloned into the pCI-neo vector to construct the recombinant eukaryotic expression plasmid,pCI-Cx26.The recombinant plasmid was identified by restriction endonuclease digestion,and transfected into COS-7 cells by liposome.The expression of Cx26 mRNA and the protein were analyzed by RT-PCR and SDS-PAGE,respectively.Results Restriction endonuclease digestion analysis verified successful construction of the recombinant plasmid,pCI-Cx26.The expression of Cx26 mRNA and protein in the transfected COS-7 cells were detected by RT-PCR and SDS-PAGE,respectively.Conclusion The eukaryotic expression plasmid for human Cx26 has been constructed successfully with the capability of expression in COS-7 cells.

Objective To investigate function of Arg327Ile (R327I) and Arg327Ala(R327A) FⅨ mutants and to study the molecular pathogenesis of haemophilia B(HB) caused by R3271 mutation.Methods Hygromycin-resistant cell line was screened and the secretion of FⅨ antigen into the medium was measured by ELISA.The cell line with appropriate expression levels of F Ⅸ antigen was selected for culture.Recombinant F Ⅸ (rF Ⅸ ) was purified from concentrated medium by two step methods of Q-Sepharose Fast Flow and anion exchange chromatography.The concentration and purity of rF Ⅸ were determined by ELISA and SDS-PAGE,respectively.The activation of wild-type ( WT),R327I and R327A of rFⅨ by FⅦa/TF/Ca2+ or FⅪa/Ca2+ was identified by Western blot in different time periods.The FⅨa and FⅧa complex formed by interaction with different concentrations of FⅧa was used to activate F X,the apparent dissociation constant (Kd) for FⅧa binding was calculated by the kinetic results.The kinetic data of the activation of FX by WT,R327I and R327A FⅨa with or without FⅧa were calculated.Results The amount of WT,R327I and R327A rFⅨ were 450,210,64 μg,and the purity of rFⅨ was confirmed by SDSPAGE.Both R3271 and R327A could be normally activated by FⅧa/TF/Ca2+ or FⅪa/Ca2+.Kd for FⅧa binding showed that the binding capacities of R327I and R327A were 4 and 5 times lower than WT,respectively.The catalytic efficiencies of R327I and R327A F Ⅸ a for F X were 6 and 8 times lower with FⅧa,and 3 and 7.4 times lower without F Ⅷ a,respectively.Conclusions R327I and R327A rF Ⅸ mutants impair their binding to the FⅧa.The site on R327 contributes to FⅧa binding.It is partly related to the activation of FX.The low FⅧa binding to R327I FⅨa may cause HB.

Objective To establish and evaluate 3 kinds of minimally-invasive valve implantation model in vivo.Methods A novel tissue engineered heart valve(TEHV) manufactured by branched polyethylene glycol cross-linked acellular porcine valve and a minimally-invasive valve implantation system according to the design of Corevalve revalving system were adopted.After anesthesia,18 adult male goats were randomly divided into 3 groups: the ulrasound-directed orthotropic group (group A,n =6),angiography-directed orthotropic group (group B,n =6) and direct-released heterotopic group (group C,n =6),and all received minimally-invasive valve implantation orthotropically or heterotopically.4 weeks later,the valvular function was evaluated by CTA and/or echocardiography.Results All 3 kinds of caprine model were successfully constructed.The operation success rate of each group was A: 66.7％,B: 50.0％ and C: 100.0％,respectively(multiple x2 analysis,group A and B P ＞0.05; group A and C,group B and C,P ＜0.05).The operation-time of each group was A: (79 ± 18) min,B:(61 ±23) min,C: (45 ± 15) min(one-way ANOVA,P ＜0.05).The survival rate at4 weeks was A: 100％,B: 100％ and C: 83.3％ (multiple x2 analysis,P ＞ 0.05).Echocardiography and CTA proved the short-term function of implanted TEHV was satisfactory.Conclusion All 3 kinds of caprine valve implantation model can be established without cardiopulmonary bypass and blood transfusion.The devices and equipments required in group A is relatively simple,but the procedure cost longer time for it is hard to determine the right position by ultrasound.The application of angiography made the positioning much easier in group B while the procedure had to be performed in specific operation room with angiographic apparatus.Group C did rely on neither special equipments nor complex operation,but the valve leaflets cannot work normally,so this model was only suitable for testing in vivo characteristics such as biocompatiblities.

The interferons(IFNs) are a family of the multifunctional cytokines, which are a kind of highly active and multifunctional glycoproteins.Studies in recent years have shown that IFNs exert a powerful antitumor effect by regulating the proliferation of tumor cells, suppressing tumor metastasis and angiogenesis, and activating antitumor immune response.Combined with other tumor treatment methods, it can inhibit the development of a variety of blood system tumors and solid tumors.In addition, the use of IFNs inducers or IFNs combined with emerging immunotherapy can significantly increase the effectiveness of tumor therapy.This review focuses on our understanding of antitumor mechanism and clinical application of IFNs, and provides some guidance for future research and clinical treatment.