Lysostaphin is highly effective on eliminating methicillin resistant Staphylococcus aureus (MRSA). In order to achieve controlled release of lysostaphin, a biocompatible drug carrier is needed. Hydroxyapatite/chitosan (HA/CS) composites were chosen to carry lysostaphin and sample composites with different weight ratios of HA to CS, including 80/20, 70/30, 60/40, and 40/60, were prepared. Multiple analyses were performed to determine the structural and physicochemical properties of the composites, including scanning electron microscopy, X-ray diffraction and Fourier transform infrared spectroscopy. We immersed HA/CS composites loaded with 1 wt% lysostaphin to test in vitro release activity and cultured MC3T3-E1 cells to carry out biocompatibility test. The result of the release behavior of the composites revealed that the controlled release of lysostaphin from 60/40 HA/CS composites was the highest release rate of (87.4 ± 2.8)%, which lasted for 120 hours. In biocompatibility testing, MC3T3-E1 cells were able to proliferate on the surface of these composites, and the extract liquid from the composites could increase the growth of the cells. These results demonstrate the controlled release of lysostaphin from HA/CS composites and their biocompatibility, suggesting the potential application of these composites to bone injury and infection applications.
3T3 Cells ; Animals ; Biocompatible Materials ; Chitosan ; chemistry ; Delayed-Action Preparations ; Drug Carriers ; chemistry ; Durapatite ; chemistry ; Lysostaphin ; pharmacology ; Materials Testing ; Methicillin-Resistant Staphylococcus aureus ; Mice ; Microscopy, Electron, Scanning ; X-Ray Diffraction

Objective To evaluate the performance of Cys C results among two detection system.Methods The particle-enhanced immunonephrometic assay was used in Dade Behring BNII. Immunoturbic assay was used in Hitachi 7170 to evaluate the JING' YUAN reagents.We compared the precison,linearity,interference,correlation,and calibrators agreement with Dade Behring BNII.Results The total CV of the samples that contain 0.6-5.0 mg/L was less than 10%.The Dade Behring and JING'YUAN method showed good linearity.Haemoglobin(10 g/L),Bilirubin(300 mg/L), Vitamin C(5 g/L)in the tested sample had no significant interference in the assay(interference 0.05) between JING' YUAN and Dade Behring reagents.Values were slightly lower than that from the Dade Behring BNII method,the mean bias was-0.16.The bias range was 1.1%-23% between JING'YUAN and Dade Behring for one sample.Conclusions The precision,linearity and interference test were suitable for routine Cys C measurement on automated biochemistry analyzer,but results has bias.

Objective To compare the coherence of serum creatinine,creatinine clearance(Ccr), Cystatin C,and estimated glomerular filtration rate(eGFR)in each stage of chronic kidney disease(CKD) patients.Methods Creatinine in serum and urine were determined by Jaffe method;serum Cystatin C was measured by particle enhanced turbidimetric method,while eGFR was calculated using the abbreviated Modification of Diet in Renal Disease(MDRD)equation which was mainly based on the serum creatinine concentration.According to the American national kidney foundation-Kidney Disease Outcome Quality Initiative(NKF-K/DOQI)guideline,all cases were grouped by eGFR into 5 stages.Results In these 228 cases,as eGFR decreased gradually,the average levels of creatinine and Cystatin C increased,while Ccr decreased.The level of each items showed a statistic difference among each stage(P0.05);in eGFR 60-89 ml/min group,the average level of creatinine was 83.3 ?mol/L,the abnormal rate was only 6.8%,it was not a sensitive marker to detect the slightly damaged GFR,the levels of Ccr and Cystatin C showed a marginal decrease and increase,with an abnormal rates of 70% and 86%,there was a statistic difference among the three abnormal rates(P

OBJECTIVETo explore the feasibility of evaluating complete ischemia-reperfusion injury (IRI) of the testis by contrast-enhanced ultrasonography with microbubbles (MB) targeted to P-selectin (MBp) in rabbits.

METHODSWe randomly divided 30 healthy adult rabbits into five groups of equal number (control, 0.5 h IRI, 1 h IRI, 2 h IRI, and 4 h IRI), prepared phospholipid MB and MBp, and performed contrast-enhanced ultrasonography of the bilateral testes with MB or MBp at an interval of 20 min at different times after IRI. When MB or MBp disappeared completely in the healthy testis at 4 to 5 min after intravenous injection, we recorded the power of the first frame (F-P) in the IRI testes followed by immunohistochemical staining of the testis tissue.

RESULTSCEU with MBp achieved a significantly higher F-P than that with MB in all the IRI groups (P < 0.05), which was (8.34 +/- 1.20) versus (1.87 +/- 0.25) 10(-5) AU at 2 hours, but there was no significant difference between MB and MBp in the control rabbits (0 AU, P > 0.05). Immunohistochemistry showed a significantly time-dependent increase in the expression of P-selectin in the vascular endothelial cells of the IRI testes, but not in those of the control.

CONCLUSIONContrast-enhanced ultrasonography with MBp can be used to evaluate the inflammatory reaction of testicular ischemia-reperfusion injury.

Animals ; Antibodies ; Disease Models, Animal ; Male ; Microbubbles ; P-Selectin ; immunology ; Rabbits ; Reperfusion Injury ; diagnostic imaging ; Testis ; blood supply ; Ultrasonography

Objective To develop a new radial artery puncture fixing device to conquer the deficiencies of traditional method. Methods The device was composed of an arm support,a hand support,a wrist part,an arm fixing belt,a palm fixing belt and a tourniquet.The arm support was concave-shaped,and was connected with the hand support with the wrist part.The wrist part had a raised block which was gifted with an inflatable bag.The balloon linked with an inflation balloon with an inflation tube.There was a valve between the inflation tube and balloon.The fixing belt had one end directly connected with the arm support, hand support and wrist part, and the other end linked with the other ends of the above components with Velcro. There was a compression hemostatic balloon at the internal surface of the tourniquet. Results The device behaved well in puncture time and success rate,and decreased the incidence rates of errhysis and hematoma after withdrawing the puncture needle while increased the satisfaction of medical staffs and patients. Conclusion The device gains advantages in simple structure,convenience,practicability and safety,and meets the desired requirements.[Chinese Medical Equipment Jour-nal,2018,39(5):44-46]

Objective To evaluate the value of using a linear stapler device for the cloure of the pharynx during total laryngectomy.Methods Sixteen total laryngectomies were performed between August 2010 and December 2011,during the operation,the TA 60 linear stapler was used for pharyngeal closure.Among these patients,two patients had the history of pre-operative radiotherapy,four patients recurred after radiotherapy,ten patients were treated for the first time.100 ml methylene blue was injected into the newly closed laryngopharyngeal cavity through the nasopharyngeal breather pipe for checking up whether it was watertight or not.Results Amnong the sixteen patients,methylene blue leakage from the mucosal joint of the gular cavity closed by the stapler were not found in fifteen patients,it was only found in one patient.The transudatory places were sutured with absorbable Vicryl sutures. This patient healed well without pharyngocutaneous fistula.Negative surgical margins were achieved in all patients.No patient needed to be tranfered to open surgery.Using a linear stapler device in total laryngectomy,45 minutes could be saved as compaired to manual suture. One patient developed a light pharyngocutaneous fistula.The incidence of pharyngocutaneous fistula was 6.25％ (1/16). Conclusions This stapled closed technique for pharyngoplasty is efficient,eliminates the risk of wound contamination,saves operation time and decreases the incidence of pharyngocutaneous fistula.This technique can be recommended as alternative for repairing the pharynx in patients undergoing total laryngectomy.

To construct a hepatitis C virus (HCV) genotype 2a monocistronic replicon and investigate its replication capabilities in the human hepatocarcinoma cell lines, Huh7.5 and Huh7.1, in order to determine its potential as a molecular tool for future in vitro studies of HCV replication and selection studies for putative anti-HCV drugs. Site-directed mutagenesis was used to delete the Core-E1-E2-p7-NS2 fragment (about 3090 bp) from plasmid pJ6JFH1BlaRL. The resultant trianglepJ6JFH1BlaRL plasmid was digested with AgeI and AvrII to release the cDNA fragment (hereafter, referred to as fragment L) containing partial 5'-untranslated region (UTR), the first 12 amino acid (aa) of HCV Core coding sequence, full-length coding sequences for the blasticidin-resistance gene, Renilla luciferase, foot-and-mouth disease virus (FMDV) 2a antiprotease and ubiquitin, and partial coding sequence for HCV NS3. To generate the monocistronic replicon, pSGRmJFH1BlaRL, fragment L was ligated into the pSGR-JFH1 vector that had been digested with AgeI and AvrII to remove the partial 5'-UTR, the first 19 aa of HCV Core coding sequence, the full-length coding sequence for the neomycin phosphotransferase II gene, the internal ribosomal entry site from encephalomyocarditis virus, and partial HCV NS3 coding sequence. A replication-defective mutant replicon, pSGRmJFH1BlaRL/GND, was constructed by a similar procedure using the pSGR-JFH1/GND vector. Fragment L was confirmed in both constructs by sequencing. Replicon RNAs were prepared from XbaI-linearized plasmid DNA templates with Invitrogen's T7 MEGAscript kit, and were purified by DNase I treatment and LiCl precipitation. RNAs were quanti?ed by optical density, and the quality and concentration were con?rmed by agarose gel electrophoresis. Replicon RNAs were transfected into Huh7.5 and Huh7.1 cells using Invitrogen's DMRIE-C transfection reagent at a ratio of 5 mug of lipid to 1mug of RNA. Time course assay of Renilla luciferase activity indicated the replicon's replication function. The pSGRmJFH1BlaRL monocistronic replicon and pSGRmJFH1BlaRL/GND replication-defective mutant replicon were successfully constructed. The pSGRmJFH1BlaRL replicon was replication-proficient in Huh7.5 and Huh7.1 cells, with replication peaking at 72 hours post-transfection and decreasing after 96 hours. No replication was detected at any time point post-transfection for the defective mutant replicon. A monocistronic replicon of HCV genotype 2a was constructed and shown to be replication-proficient in human hepatocarcinoma cell lines.
Cell Line, Tumor ; Genetic Vectors ; Genome, Viral ; Genotype ; Hepacivirus ; genetics ; Humans ; Mutagenesis, Site-Directed ; RNA, Viral ; Transfection ; Virus Replication

OBJECTIVETo study the epidemiologic characteristics of hand-foot-mouth disease (HFMD) in Guiyang between 2008 and 2010.

METHODSThe epidemiologic characteristics of HFMD were analyzed by descriptive statistical methods based on the data from the China Information System for Disease Control and Prevention.

RESULTSA total of 27383 cases of HFMD were recorded in Guiyang between 2008 and 2010. The incidence of HFMD increased from 66.4439/100000 in 2008 to 163.9276/100000 in 2009 and 471.5515/100000 in 2010 (P<0.01). The mortality rate was 0.1026/100000 in 2010, which was significantly lower than in 2009 (0.2821/100000) (P<0.05). HFMD occurrence showed seasonality and reached a peak between April and June. HFMD cases were commonly noted in children under 5 years old, and especially in children under 3 years old. The main detected pathogen was human enterovirus 71 (EV17) in 2009. Whereas in 2010 the disease was mainly caused by CoxA16 and other intestinal viruses.

CONCLUSIONSThe incidence of HFMD in Guiyang increased year by year from 2008 to 2010, but the mortality rate decreased year by year. HFMD occurrence showed an obvious seasonality. HFMD was common in children under the age of five. The main pathogens of this disease included EV17, CoxA16 and other intestinal viruses.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Enterovirus A, Human ; isolation & purification ; Hand, Foot and Mouth Disease ; epidemiology ; virology ; Humans ; Infant ; Middle Aged ; Time Factors

Objective Some research has shown that prolactin (PRL) is closely related to severity of hypoxic-ischemic encephalopathy (HIE). However, the role of maternal and neonatal plasma PRL levels in neonatal asphyxia has not been reported so far. This paper aims at studying the changes of PRL levels in the cord blood, maternal blood and plasna of newborns in neonatal asphyxia. Methods The maternal blood, cord blood and neonatal plasma PRL levels in 25 neonates with asphyxia (asphyxia group) and 20 normal ones (control group) were detected by radioimmunoassay.Results The maternal blood, cord blood and neonatal plasma PRL levels [(362.5 + 127.1), (984.6 + 262.3) and(386.3+216.2) μg/L, respectively] in the asphyxia group were significantly higher than those in the control group[(96.4+26.2), (92.3+ 18.4) and (68.7+7.27) μg/L, respectively] ( P ＜0.01). The maternal blood, cord bloodand neonatal plasma PRL levels [ (445 + 216), (996 + 284) and (412 + 221) μg/L, respectively] in the severe asphyxia groupwere higher than those in the mild asphyxia group [(298 + 102), (612 + 221) and (309 + 19.2) μg/L,respectively] ( P ＜0.01 or 0.05). The cord blood and neonatal plasma PRL levels had a positive correlation both in the mild and the severe asphyxia group ( r = 0.54, r = 0.63 , both P ＜ 0.05 ). The plasma PRL level right after resuscitation was higher than that of the control group ( P ＜0.01). It gradually reduced from the 2nd day after birth,but was higher than that of the control group ( P ＜0.01). The PRL level on the 10th day after birth was not different from that of the control group. Conclusions The PRL levels of neonatal plasma, cord blood and maternal blcod increase in the perinatal asphyxial newborns. The plasma PRL level may be a good marker to evaluate the degree of asphyxia.

OBJECTIVETo preliminarily investigate the relationship between the Lactate dehydrogenase (LDH) activity of Streptococcus mutans (S. mutans) and dental caries initiation.

METHODS100 S. mutans strains derived from caries-active and caries-free individuals were cultured in BHI medium supplemented with glucose. Cells were extracted and ruptured, and the extracted liquid protein was quantified with Coomassie brilliant blue G250 staining methods. LDH activity was assayed using the pyruvate-dependent oxidation of NADH-with and without FDP.

RESULTSLDH activity of the two groups strains had no difference (P > 0.05), but the distribution of differ class enzyme activity strains in the two groups was different (P < 0.05).

CONCLUSIONLDH activity of S. mutans is correlated to the initiation of dental caries to some extent. The measurement methods in this study can be applied in preliminary quantitation of LDH activity and the screening of S. mutans strains.

Case-Control Studies ; Dental Caries ; microbiology ; Humans ; Lactate Dehydrogenases ; Oxidation-Reduction ; Streptococcus mutans ; metabolism