Objective:To compare the optimal conditions, virus yield, viral titer and cell metabolism between culturing influenza virus H1N1 vaccine strain in MDCK and MDCK-G1 cells.Methods:The optimal culture conditions were investigated using chessboard method. The hemagglutination titer, half of the tissue infection dose (TCID 50) and the metabolism of glucose and lactic acid were monitored and compared between the two cell lines. Results:After MDCK-G1 cells were inoculated with H1N1 at the multiplicity of infection (MOI) of 0.001 with the presence of 1 μg/ml of trypsin, the hemagglutination titer reached the peak of 1∶512 at 72 h and the viral titer was 10 7.4TCID 50/ml. In the MDCK cell line group, the hemagglutination titer reached the peak of 1∶256 at 72 h and the viral titer was 10 6.6TCID 50/ml when using H1N1 at MOI=0.0001 and 1 μg/ml of trypsin. Conclusions:MDCK-G1 cells were more suitable than MDCK cells for the proliferation of influenza virus. This study provided reference data for further research on cell-derived influenza vaccine.

Objective: To investigate the best amount of TPCK trypsin in Madin Darby canine kidney (MDCK) cell suspension for the culture of H7N9 avian influenza virus. Methods: Different concentrations of TPCK trypsin were added during the periods of cell growth and virus production. Their effects on cell growth, viability, glucose and lactate metabolism, and hemagglutination titer were monitored every 12 h. Inter-batch differences were analyzed. The amount of trypsin added in the cell growth phase was 0, 1 μg/ml, 2 μg/ml, 4 μg/ml, 6 μg/ml, 8 μg/ml, 10 μg/ml and 15 μg/ml. The amount of trypsin added during the virus production period was 0, 0.5 μg/ml, 1 μg/ml, 1.5 μg/ml, 2 μg/ml and 2.5 μg/ml. When the hemagglutination titers were same, the adding amount was further optimized at different multiplicity of infection (MOI) of 0.001, 0.005, 0.025 and 0.05. Results: No significant linear effects of TPCK trypsin concentration on cell number, viability, and glucose and lactate metabolism were observed. No toxicity to cell growth was observed when TPCK trypsin concentration reached 15 μg/ml. After the inoculation of H7N9 avian influenza virus, the hemagglutination titers in the 1 μg/ml, 1.5 μg/ml, 2 μg/ml and 2.5 μg/ml TPCK trypsin groups reached the peaks at 48 h, which were 1∶2^6.5. At 60 h, the hemagglutination titers of the latter two groups decreased faster than those of the former two groups. When the MOI was 0.005, the hemagglutination titer of the 1.5 μg/ml group at 48 h was 2^6.5 higher than 2⁶ in the 1 μg/ml group under the same condition. There were differences between different batches of TPCK trypsin. Conclusions Adding 1 μg/ml and 1.5 μg/ml of trypsin could better promote the proliferation of H7N9 avian influenza virus, and 1.5 μg/ml of trypsin had a wider range of MOI applicability.

Objective To evaluate the efficacy of interventional therapy via apical approach in complex mitral valve paravalvular leakage (PVL).Methods From January 2014 to December 2016,interventional therapy by apical approach was used in 7 patients with mitral valve PVL after mechanical valve replacement.There were 5 male and 2 female with mean age of 42-64 (51.3 ± 7.1) years.Six patients had a history of previous infective endocarditis.There were 3 cases of NYHA heart function =Ⅲ,and 4 cases =Ⅳ.The period of time between interventional therapy and previous operation was 6.5-8 (3.6 ± 3.1) years,with mitral regurgitation volume:9.5-23.1 (13.3 ± 4.7)ml.Interventional therapy of small incision method via the left sixth intercostal was carried out in the catheterization laboratory or the hybrid operation room with the patient under general anesthesia.Follow-up evaluation included peri-operational mortality,complications,improvement of cardiac function,hemolysis and postoperative residual mitral regurgitation.Results The success rate of total operation was 100％.The average operation time was 90-300 (145.7 ± 71.8) min,and the DSA radiation time was 6-25 (12.1 ± 6.5)min,with average hospitalization time of 5-12 (10.2 ± 3.5)d.The main post-operative complications included 1 case of hemoptysis,1 case of hematuria and acute renal failure,and 2 cases of blood transfusion,with blood transfusion volume of 1 200 ml and 3 290 ml,respectively.During the follow-up,there was no death.Mitral regurgitation volume decreased to 0 -1.0(0.43 ± 0.45) ml (P ＜ 0.05).All patients had improved heart function in different degree and no serious complications.Conclusions Interventional therapy via apical approach in complex mitral valve PVL has the advantages of being a simpler and safer approach,less trauma,shorter time of hospitalization,faster post-operative recovery and lower treatment cost.Its disadvantages are high difficult operation,strict indications and high technical requirements for the surgeon.By improving operation skills,choosing individualized occluders and treatment of early hemolytic complications,the success rate and long-term effect of interventional therapy can get further improved.

BACKGROUND: Irradiation crosslinking can remarkably increase the abrasive performance of ultra-high molecular weight polyethylene (UHMWPE), thereby reducing its wear rate. OBJECTIVE: To review the recent progress in irradiated crosslinking UHMWPE. METHODS: A computer-based online search of ISI Web of knowledge was performed for articles related to irradiated crosslinking UHMWPE published from January1995 to April 2012 using the keywords of“irradiation crosslinking UHMWPE” in English. RESULTS AND CONCLUSION: At present, domestic studies on irradiation crosslinking of UHMWPE mainly focus on the wear resistance, oxidation resistance and mechanical properties. Under high-energy radiation, radicals generate from the UHMWPE, which are inter-crosslinked. Irradiation crosslinking dramatical y enhances the UHMWPE wear resistance, while reduces its oxidation resistance and mechanical properties. In the future, the research wil focus on how to develop new UHMWPE composite with good wear resistance, oxidation resistance and mechanical properties.

OBJECTIVETo assess the association of three polymorphisms (14-bpINS/DEL, +3035C/T and +3142C/G) in the 3' untranslated region (3'UTR) of HLA-G gene and systemic lupus erythematosus (SLE) in Yunnan.

METHODSA case-control study has been carried out on 206 SLE patients and 212 healthy controls. Genotypes of 14-bpINS/DEL (rs1704), +3035C/T (rs17179108) and +3142C/G (rs1063320) loci of 3'UTR of the HLA-G gene were determined with DNA sequencing.

RESULTSAllelic and genotypic frequencies of 14-bpINS/DEL and +3142C/G did not differ significantly between the two groups (P > 0.05). The frequencies of +3035T allele was significantly higher in the SLE group compared with the control group (P < 0.01, OR=1.604, 95% CI: 1.186-2.169). With a dominant inheritance model, the odd ratio of dominant genotype (CT+TT) was 2.004 (95% CI: 1.345-2.987, P=0.0006) in the SLE group.

CONCLUSIONThe 14-bpINS/DEL and +3142C/G polymorphisms in the 3'UTR of the HLA-G gene are not associated with susceptibility to SLE in Yunnan, whilst the T allele of +3035C/T may be a risk factor for SLE.

3' Untranslated Regions ; genetics ; Case-Control Studies ; China ; Female ; Genetic Predisposition to Disease ; HLA-G Antigens ; genetics ; Humans ; Lupus Erythematosus, Systemic ; genetics ; Male ; Polymorphism, Genetic

Objective To study the feasibility of modified early warning scores (MEWS) for assessing the severity and death prediction in the pre-hospital traumatic patients.MethodsData of the prehospital traumatic patients admitted between January 1,2010 and December 31,2010 were collected and assessed onsite by using MEWS.Numeration data was presented in percentage by using chi-square test,and measurement data was xepressed in mean with standard deviation,and P ＜ 0.05 was considered to be difference with statistical significance.Observation was lasted for 90 days after admission to get final results as observation object and ROC curve was drew and calculated the area under the curve for predicting severity and death of patients.The patients without vital signs and unsuccessful resuscitations were not included in this study.ResultsThere were 1475 (87.95％) cases/times with score of 0 -2,and 202 (12.05％)cases/times with score of 3 - 13.In the non-survival group,MEWS were higher than that in the survival group with statistic significance ( P ＜ 0.01 ).When the area under ROC was 0.94,the optimal cutoff point for potentially severe patients was MEWS≥3 for predicting the death of severe pre-hospital traumatic patients with sensitivity of 85.7％,specificity of 88.6％,accuracy of 88.6％ and Youden of 0.743,showing high significance of the application of MEWS to assessing severity of traumatic patients and death prediction.ConclusionsMEWS used to assess the pre-hospital traumatic patients and predict death with high validity and accurate quantification is a simple,practical and easily operable method with strong application significance.

OBJECTIVETo explore the relationship between genetic polymorphisms of 3 single nucleotide polymorphisms (SNPs) in the elastin microfibril interfacer 1 (EMILIN1) gene and essential hypertension.

METHODSA case-control study was conducted in which 201 hypertensive patients and 202 healthy controls in Mongolian population were enrolled, and the genotypes of rs3754734, rs2011616 and rs2304682 loci were analyzed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct sequencing techniques.

RESULTSThere were significant differences in the frequencies of alleles and genotypes for the rs2304682 between the hypertensives and normotensives in the population (P<0.05). The frequency of the G-G haplotype established by rs3754734 and rs2304682 was significantly higher in the hypertensive patients (P<0.05). The frequencies of alleles and genotypes for the rs2304682 also had significant differences between the group with high diastolic blood pressure and normal diasto lic blood pressure (P<0.05).There were no significant differences in the frequencies of alleles and genotypes for the 3 SNPs between the group with high systolic blood pressure and normal systolic blood pressure (P>0.05).

CONCLUSIONThe rs2304682 locus in the EMILIN1 gene, as well as the haplotypes G-G constructed using rs3754734 and rs2304682, may associate with the susceptibility of essential hypertension in the Mongolian population. Also, rs2304682 may associate with the level of the diastolic blood pressure.

Blood Pressure ; genetics ; Case-Control Studies ; Genetic Predisposition to Disease ; Humans ; Hypertension ; genetics ; Membrane Glycoproteins ; genetics ; Middle Aged ; Mongolia ; Polymorphism, Single Nucleotide

Objective To investigate the peri-nasopharyngeal invasion patterns of nasopharyngeal carcinoma (NPC) on MRI and its relationship with tumor staging. Methods One thousand five hundred and seventy-three patients with newly diagnosed NPC which were histo-pathologically proved were retrospectively studied. The MRI manifestations and invasion patterns of the NPCs were elevated according to the 2008 Tstaging system of NPC. Z test was used to analyze the rate of adjacent structures invasion in NPCs. Results The structures invaded by NPCs included pharyngobasilar fascia in 1299 cases (82. 58% ); parapharyngeal space, 1090 ( 69. 29% ); nasal cavities, 304 ( 19. 33% ); oropharynx, 49 ( 3. 12% ); carotid space,514(32. 68% ); medial pterygoid muscle, 661 (42. 02% ); lateral pterygoid muscle, 210( 13. 35% ); skull base bones, 943(59. 95% ); cranial nerves, 630(40. 05% ) and paranasal sinuses, 242 ( 15.38% ). The T-stage distribution was T1, 242 cases ( 15.38% ); T2, 288 ( 18. 31% ); T3, 410 (26. 06% ) and T4,633 (40. 24% ). Among the cases with nasal cavities invasion, 90. 46% (275/304)showed the involvement of the structures seen in T3 or T4 stage, which was found in all cases with oropharynx invasion. In addition,69. 14% (457/661) of cases with medial pterygoid muscle invasion and 92. 15% (223/242) of cases with paranasal sinuses invasion showed the involvement of structures seen in T4 stage. As for the invasion patterns of NPC, the lateral invasion of pharyngobasilar fascia was more frequent than upward invasion of skull base (Z = 14. 025, P ＜ 0. 01 ) and downward invasion of oropharynx ( Z = 45.032, P ＜ 0. 01 ), and the downward invasion of oropharynx was less frequent than upward invasion of skull base ( Z = 34. 301, P ＜ 0. 01 ) and forward invasion of nasal cavities ( Z = 14. 404, P ＜ 0. 01 ). Conclusion NPC has a predilection of lateral invasion rather than upward and downward invasion, and its upward and forward invasion are more common than downward invasion.

BACKGROUND: How to make growth factor plays a role persistently and efficiently is a key in constructing bone tissue engineered bone. Currently, varied microspheres or scaffolds were used as release carriers, however, the delayed release effects needs elevating.OBJECTIVE: To prepare chitosan microosPheres/nano-hydroxyapatite/poly (lactic-co-glycolic acid) (CMs/nHA/PLGA) scaffolds, and to measure its characteristics of delayed release of bovine serum albumin (BSA).METHODS: CMs were prepared by an emulsifying cross linking method with BSA as a model protein. Using ice particulates as porogen, composite CMs/nHA/PLGA scaffolds were prepared by freeze-drying. The characteristic and morphology of the composite were observed by scanning electron microscope, later particle size analyzer, mercury porosimeter and universal testing machine, and the release behavior of BSA was investigated in vitro.RESULTS AND CONCLUSION: The CMs were spherical shape with a regular surface, with diameters of 20-40 μm. The encapsulation efficiency of the CMs was 86.5%, and the loading capacity was 0.8%. With the increase of initial BSA dosage, the loading capacity increased to 2.6%, while the encapsulation efficiency decreased to 74.1%. The CMs can be uniformly distributed in PLGA scaffolds to form CMs/nHA/PLGA scaffolds, which had 100-400 μm pore diameter and over 80% porosity, with 1.1-2.3 pMPa compressive strength, and 26.5% cumulative degradation at 10 weeks. The cumulative release of BSA from nHA/PLGA scaffolds was above 85% at 36 hours, which from CMs was 33.6% at 10 days, and that from CMs/nHA/PLGA scaffolds was 81.5% at 40 days. The results demonstrated that CMs/nHA/PLGA scaffolds have an excellent releasing efficiency for protein drugs with suitable compressive strength and degradation, which would be used as delivery system and tissue engineering scaffolds.

OBJECTIVE: To study the expression of apoptosis related genes, Bcl-2, bax, and iNOS in the olfactory epithelium of mice infected with influenza virus, and to discuss how they regulate apoptosis of the olfactory sensory neurons. METHOD: The expression levels of apoptosis related genes were detected with semi-quantity RT-PCR. RESULT: (1) The expression levels of Bcl-2 mRNA remain relatively constant after virus inoculation; (2) The expression levels of bax mRNA increased massively, and decreased gradually; (3) The expression levels of iNOS mRNA increased significantly in a short period, and then fell down to the undetectable level gradually. CONCLUSION The apoptosis related genes, Bcl-2, bax, and iNOS may play important roles in regulation of apoptosis of olfactory sensory neurons of mice infected with influenza virus.
Animals ; Mice ; Mice, Inbred BALB C ; Nitric Oxide Synthase Type II ; metabolism ; Olfactory Mucosa ; metabolism ; Orthomyxoviridae ; Orthomyxoviridae Infections ; metabolism ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; bcl-2-Associated X Protein ; metabolism