To construct secretory expression vector of PPV NS1 gene, the fragment of PPV NS1 gene coding for major antigen region of the NS1 protein was amplified by PCR and inserted into multiple clone site of eukaryotic expression vector pPICZalpha-A. The recombinant pPICZalpha-A-NS1 plasmid was transferred into P. pastoris strain GS115 mediated by electro transform. Recombinant P. pastoris strain GS115 was induced to express the fusion protein by methanol. The expressed and purified protein was analyzed by SDS-PAGE and Western Blot. The recombinant protein was highly-expressed and showed a good immunoreactivity. The indirect ELISA method was developed for detecting antibodies against PPV by checkerboard titration assay. The result showed that the optimal concentration of coated antigen was 3.2 microg/mL and the best dilution of serum was 1 : 80. The positive cut-off value of the ELISA assay was OD450 > 0.4 and OD450 positive serum/OD450 negative serum > 2.0. Compared with HI and commercial ELISA kits, the assay revealed 94.2% and 92.1% agreement respectively. The assay demonstrates good specificity and sensitivity, and can be applied in the detection of porcine parvovirus.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Parvoviridae Infections ; diagnosis ; immunology ; veterinary ; virology ; Parvovirus, Porcine ; genetics ; immunology ; isolation & purification ; Recombinant Proteins ; genetics ; immunology ; Swine ; Swine Diseases ; diagnosis ; immunology ; virology ; Viral Nonstructural Proteins ; genetics ; immunology

·AIM: To analyze the relationship between tear film break-up time (BUT) and systemic factors. ·METHODS: From January 2015 to July 2016 in the medical examination center of Fuzhou General Hospital or hospitalized, there were 747 cases included. All participants were observed for systolic and diastolic blood pressure, blood glucose, total cholesterol, high density lipoprotein cholesterol,low density lipoprotein,break-up time equivalent. The relationship between tear film rupture time and systemic factors was analyzed with simple and multiple linear regression analysis by SPSS19.0. ·RESULTS: Simple linear regression analysis results showed that the diastolic pressure and tear break-up time were positively correlated(P<0.05);age,systolic blood pressure, blood glucose, cholesterol, high density lipoprotein, low density lipoprotein was negatively correlated with BUT (P < 0. 05), but no correlation between triglyceride and tear film break up time (P = 0.227). Multiple linear regression analysis suggested that the tear film rupture time was positively correlated with diastolic blood pressure (P<0.001), but negatively correlated with systolic blood pressure (P<0.001), blood glucose (P<0. 001) and high-density lipoprotein (P=0.019). The time of tear film rupture was not related to age, cholesterol, triglyceride, low density lipoprotein.·CONCLUSION: The correlation between blood glucose and tear film BUT is the most significant among all the indexes involved, indicating that ocular surface damage may occur in diabetic patients.

BACKGROUND:As a noticeable tissue engineering material of polyhy droxyalka noates family, poly (3-hydroxybutyrate-co-4-hydroxybutyrate)(P3HB4HB) exhibitsgood biocompatibility, adhesion and mechanicalproperties, presenting aextensive application future in tissue-engineered research.
OBJECTIVE:To investigate the biocompat ibilityin vitroand ectopic osteogenic differentiationin vivoof P3HB4HB and human bone marrow mesenchymal stem cels.
METHODS:Passage 5human bone marrow mesenchymal stem cels transplanted ontothe three-dimensional P3HB4HB scaffoldwereincubated with osteogenic induction medium (test group)or with no osteogenic induction(control group), respectively. After 5-day incubation, thecelgrowth was assessed by acridine orange staining and scanning electron microscopy; after14-day incubation, both kinds of cel-scaffold composites were subcutaneously implanted into the nude mice. At 16 weeks after implantation, the cel-scaffold composites were removed to observeectopic osteogenic differentiationin vivousing hematoxylin-eosin staining, von Kossa staining and colagen type I immunohistochemical staining.
RESULTS AND CONCLUSION:Acridine orange staining showed that cels adhered wel on the surface of the scaffold;under thescanning electron microscope, induced celsgrew wel on the P3HB4HB scaffold and produced abundant extracelular matrixes. In addition, at 16 weeks after implantation, there were osteoidtissues in the test group, positive for von Kossa staining as wel as colagen type I immunohistochemical staining;furthermore, hematoxylin-eosin staining showednumerous osteoblasts and bone lacunas. In contrast, no bone tissues appeared in the control group. To conclude, P3HB4HB is a suitable material for bone tissue engineering.

A male patient of 26 years old was received with multiple fracture by tranffic accident. Anesthesia was maintained with inhalation of isoflurance. Malignant hyperthermia (MH) was appeared after operation. The patient rehabilited after physical cooling, bedside persistence hematodialysis, hormone application and so on.
Anesthetics, Inhalation ; Humans ; Male ; Malignant Hyperthermia

Objective:This study aimed to construct a lentiviral expression vector for microRNA-194 and investigate its effect on the metastasis of human osteosarcoma cell line U2-OS. Methods:Pri-and mature miR-194 amplified by PCR were inserted into the plenty-GFP vector and identified by restriction endonuclease digestion and nucleotide sequencing. The osteosarcoma cell line U2-OS was transfected with the lentivirus. Then, the stable transfected cells were used in Transwell and wound healing assay. Results:Restric-tion analysis and sequencing showed that the recombinant lentiviral expression vector was constructed correctly. The titers of obtained overexpression and suppression expression recombinant lentivirus were 1.5*108 and 4*108 TU/ml. Cell metastasis ability was signifi-cantly different in different experimental groups (P<0.01). Conclusion:The lentiviral expression vector for microRNA-194 was suc-cessfully constructed. MicroRNA-194 could influence the metastasis of the osteosarcoma cell line U2-OS;thus, it could be further ex-plored as a potential target in osteosarcoma therapy.

OBJECTIVE: To clone the transmembrane (TM) domain sequence of EGFR gene and lay a good foundation for constructing the transmembrane expression vector of recombinant superantigens and cytokines. METHODS: A pair of primers special to the sequence encoding TM domain of EGFR gene were synthesized, TM domain fragment was cloned by RT-PCR, and the PCR product of TM domain sequence was ligated with the pGEM-T vector and confirmed by DNA sequencing. RESULTS: TM domain sequence was successfully cloned and verified by DNA sequencing. CONCLUSION: The successful cloning of TM domain sequence provides a basis for the construction of transmembrane fusion protein of Superantigen-TM or Cytokines-TM in cancer biotherapy.

Objective To analyze and summarize the clinical manifestations,diagnosis and treatment of pure spinal epidural cavernous hemangiomas.Methods We presented a case series of 6 patients with pure spinal epidural cavernous hemangiomas in our hospital from April 2012 to November 2016 with previous articles discussing their clinical presentation,radiological characteristics,surgical technique,pathological features,and functional outcome.Results All patients were diagnosed as pure spinal epidural cavernous hemangiomas by pathology.The cavernous hemangiomas were totally cut and patients recovered well after operations.All patients gradually improved neurologically and achieved a good outcome with no recurrence during the follow-up for 13 to 68 months.Conclusion Pure spinal epidural cavernous hemangioma is rare.It is easily confused with other spinal diseases.MRI is an important imaging examination method.Pathology is the method of diagnosis.Surgical resection is the main method of treatment,and the prognosis is good.

PURPOSE: Physician empathy is a core attribute in medical professionals, giving better patient outcomes. Medical school is an opportune time for building empathetic foundations. This study explores empathy change and focuses on contributory factors. METHODS: We conducted a cross-sectional study involving 881 students (63%) from Years 1 to 5 in a Singaporean medical school using the Jefferson Scale of Physician Empathy-Student version (JSPE-S) and a questionnaire investigating the relationship between reported and novel personal-social empathy determinants. RESULTS: Empathy declined significantly between preclinical and clinical years. Female and medical specialty interest respondents had higher scores than their counterparts. Despite strong internal consistency, factor analysis suggested that the JSPE model is not a perfect fit. Year 1 students had highest Perspective Taking scores and Year 2 students had highest Compassionate Care scores. High workload and inappropriate learning environments were the most relevant stressors. Time spent with family, arts, and community service correlated with higher empathy scores, whilst time spent with significant others and individual leisure correlated with lower scores. Thematic analysis revealed that the most common self-reported determinants were exposure to activity (community service) or socialisation, personal and family-related event as well as environment (high work-load). CONCLUSION: While the empathy construct in multicultural Singapore is congruent with a Western model, important differences remain. A more subtle understanding of the heterogeneity of the medical student experience is important. A greater breadth of determinants of empathy, such as engagement in arts-related activities should be considered.
Art ; Cross-Sectional Studies ; *Education, Medical, Undergraduate ; *Empathy ; Factor Analysis, Statistical ; Family ; Female ; Humans ; Male ; Medicine ; *Physician-Patient Relations ; Physicians ; Self Report ; Sex Factors ; Singapore ; Social Participation ; *Students, Medical ; Workload

This study was purposed to investigate the relationship between HLA-A, B allele polymorphisms and red blood cell parameters of patients with --(SEA/αα) subtype of α(0)-thalassemia in Han ethnic population of Wuzhou city. The HLA genetic polymorphisms were determined by polymerase chain reaction-sequence-based typing (PCR-SBT) in 57 patients with --(SEA/αα) subtype of α(0)-thalassemia of Han ethnic population in Wuzhou city, Guangxi province, China. Mean corpuscular volume (MCV), hemoglobin (Hb), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) were detected by automatic blood analyzer system. HbA2 were detected by electrophoretic method. The statistical analysis was performed by ordinal polytomous logistic regression. The results showed that Hb and HbA2 levels were significantly lower in patients positive for HLA-A*33:03, B*15:01 or B*55:02, and were significantly higher in patients positive for B*15:02 (P < 0.05). It is concluded that several HLA alleles may be associated with Hb level of --(SEA/αα) subtype of α(0)-thalassemia of Han ethnic population in Wuzhou city. This result has the value for understanding phenotype-genotype relationships in thalassemia.
Adolescent ; Adult ; Alleles ; Child ; Child, Preschool ; China ; epidemiology ; Erythrocytes ; cytology ; Ethnic Groups ; genetics ; Female ; Genotype ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; Hemoglobins, Abnormal ; genetics ; Humans ; Male ; Middle Aged ; Young Adult ; alpha-Thalassemia ; blood ; classification ; epidemiology ; genetics

OBJECTIVETo summarize the surgical effect and clinical application value of esophagectomy with extended 2-field lymph node dissection for patients with esophageal carcinoma.

METHODSFrom June 1987 to December 2008, 1690 patients with esophageal cancer underwent esophagectomy with extended 2-field (thoracic and abdominal) dissection of lymph nodes. Patients with the middle and lower thoracic esophageal cancer underwent Ivor-Lewis esophagectomy, and patients with upper thoracic esophageal cancer underwent Akiyama esophagectomy. 2-field (thoracic and abdominal) lymph node metastases information and the 1, 3, 5, 10-year survival rates were analyzed retrospectively.

RESULTSLymph node metastases were found in 713 patients. The lymph node metastases rate was 42.2% (713/1690).Thoracic lymph node metastasis rate was 39.3% (665/1690), among which in the right pleural apical para-tracheal triangle was 20.7% (349/1690), in the posterior upper mediastinum was 26.3% (444/1690), in the lower mediastinum was 18.2% (307/1690). Abdominal lymph node metastasis rate was 20.1% (339/1690). THE Postoperative complication rate was 16.4% (278/1690), among which the pulmonary complication rate ranking the first, was 43.6% (136/312). The operative mortality rate was 0.2%. The 1-year, 3-year, 5-year and 10-year survival rates were 88.2% (1388/1574), 63.5% (868/1367), 54.8% (705/1287) and 30.8% (232/754), respectively. The 5-year survival rate in patients without lymph node metastasis was 76.2% (448/588), but that in patients with lymph node metastases was 36.8% (257/669).

CONCLUSIONThe results of this study demonstrated that Ivor-Lewis and Akiyama esophagectomy with two-field lymph node dissection exposes the operation fields clearly and make radical lymphadenectomy thoroughly, especially the lymph nodes in the posterior upper mediastinum around the recurrent laryngeal nerve and in the right pleural apical para-tracheal triangle. It is essential that patients with esophageal carcinoma with lymph node metastases should undergo esophagectomy with extended 2-field dissection of lymph nodes. This can elevate the postoperative 5-year survival rate remarkably.

Adenocarcinoma ; mortality ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; mortality ; pathology ; surgery ; Esophageal Neoplasms ; mortality ; pathology ; surgery ; Esophagectomy ; adverse effects ; methods ; Female ; Humans ; Lymph Node Excision ; adverse effects ; methods ; Lymphatic Metastasis ; Male ; Middle Aged ; Respiratory Insufficiency ; etiology ; Retrospective Studies ; Survival Rate