This study was designed to explore the RNA interference technique in inhibition of the expression of the mouse fibrinogen like protein 2 (mfgl2), which has been reported to be involved in the development a variety of diseases including fulminant viral hepatitis. A plasmid named p-mfgl2shRNA,complementary to the sequence of mfgl2 was constructed, while another short hairpin RNA (shRNA)which was a mutated form of the mfgl2shRNA sequences was used as a control. A plasmid named pEGFP-mfgl2 expressing the mfgl2-EGFP fusion protein was also constructed for the screening of the effect of p-mfgl2shRNA on mfgl2 expression. By cotransfection of p-mfgl2shRNA and pEGFP-mfgl2 or pcDNA3.1-mfgl2 expression construct into CHO cells or HeLa cells, the inhibition of mfgl2 expression by mfgl2shRNA was analyzed by direct observation through fluorescent microscopy, FACS, RT-PCR and immunohistochemistry staining. The experiments showed the significant inhibitory effect of p-mfgl2shRNA on mfgl2 expression at 48h post-transfection in both CHO and Hela cell lines with the inhibitory efficiency as high as 80.1%. The study demonstrated that the construct of p-mfgl2shRNA successfully interfered with the mfgl2 expression in vitro.

This study using maltodextrin as raw material, 1%-5% polyvinylpyrrolidone K30 as template agent, 1%-5% ammonium bicarbonate as pore-forming agent, curcumin and ibuprofen as model drugs. Porous maltodextrin was prepared by template and pore-forming agent methods, respectively. The structure and drug delivery behavior of porous maltodextrin prepared by different technologies were comprehensively characterized. The results showed that the porous maltodextrin prepared by pore-forming agent method had larger specific surface area (6.449 4 m²·g^-1) and pore size (32.804 2 nm), which was significantly better than that by template agent method (3.670 2 m²·g^-1, 15.278 5 nm). The adsorption kinetics between porous maltodextrin prepared by pore-forming agent method and curcumin were suitable for quasi-first order adsorption kinetic model, and that between porous maltodextrin and ibuprofen were suitable for quasi-second order adsorption kinetic model. While the adsorption kinetics between porous maltodextrin prepared by template agent method and two model drugs were both suitable for the quasi-first order adsorption kinetic model. In addition, the dissolution behavior analysis showed that the porous maltodextrin prepared by the two technologies can significantly improve the dissolution behavior of insoluble drugs, and the drug release was both carried out by diffusion mechanism, which suitable for the Peppas kinetic release model, but the porous maltodextrin prepared by template agent method had a faster release rate. The change of nozzle diameter had no significant effect on the adsorption process and drug release behavior of porous maltodextrin. In conclusion, the porous maltodextrins prepared by two different technologies were both beneficial to the delivery of insoluble drugs, and the template agent method was the best for delivery of insoluble drugs. This study can provide theoretical basis for the preparation of porous particles, promote the application of porous particles in insoluble drugs, and improve the bioavailability of insoluble drugs.

OBJECTIVETo investigate the tumor-associated antigen CA125 expression in the serum and cervical and vaginal secretions in women during normal reproductive period, and explore the clinical value of detecting tumor markers in the cervical and vaginal secretions.

METHODSA total of 145 women in reproductive period were divided into 3 age groups (20-29 years, 30-39 years, and over 40 years), and their CA125 levels in cervical secretion, vaginal secretion and serum were detected by automatic electro-chemiluminescent immunoassay.

RESULTSCA125 levels in the cervical secretion, vaginal secretion and serum showed no significant difference between the 3 age groups (P>0.05). In each group, CA125 levels differed significantly between the cervical secretion, vaginal secretion and serum (P<0.001). In the 145 women, the average CA125 level was 497.82 - or + 75.29 U/ml in the cervical secretion, 114.66 - or + 26.40 U/ml in vaginal secretion and 18.06 - or + 3.35 U/ml in serum, showing significant differences between them (P<0.001).

CONCLUSIONCA125 expression level is significantly higher in the cervical and vaginal secretions than in the serum in women in normal reproductive period, and its levels in cervical and vaginal secretions can be more sensitive and convenient for early detection of related diseases.

Adult ; Biomarkers ; analysis ; CA-125 Antigen ; blood ; metabolism ; Cervix Mucus ; metabolism ; Female ; Humans ; Middle Aged ; Vagina ; secretion ; Young Adult

OBJECTIVETo compare the clinical effects of Cookgas intubating laryngeal airway (CILA) in facilitating fiberoptic bronchoscope (FOB) and Shikani optical stylet (SOS)-guided intubations in anticipating difficult tracheal intubation.

METHODSTotally 60 anticipated difficult tracheal intubation patients undergoing selective plastic surgery under general anesthesia were allocated to FOB group (n = 30) and SOS group (n = 30). After anesthesia induction and CILA insertion, the patients were treated with FOB or SOS-guided intubation via CILA. The time of intubation and CILA removal and the time and the success rate of CILA insertion were recorded. Noninvasive blood pressure and heart rate were recorded before and after anesthesia induction at CILA insertion, at intubation, at CILA removal, and every minute thereafter for 5 minutes.

RESULTSCILA was inserted successfully in all patients. The first intubation attempt succeeded in all but two who succeeded in the second and the third attempt respectively in FOB group. In SOS group, 18 patients were successfully intubated in the first attempt, and 7 patients were successfully intubated in the second attempt; SOS failed in 5 patients with severe cervical scars, and then FOB was successfully used to intubate. The time of the intubation [(60.2 +/- 29.6) vs. (92.4 +/- 47.9)s] and CILA removal [(104.6 +/- 39.9) vs. (130.0 +/- 51.9) s] in SOS group were significantly longer than in FOB group (P < 0.05). Hemodynamic changes during the intubation with CILA in these two groups were minimal.

CONCLUSIONSFOB and SOS-guided tracheal intubation via CILA is safe and effective in anticipating the outcome of difficult airway management. Compare to SOS-guided intubation, the time of FOB-guided intubation is shorter and the success rate is higher.

Anesthesia, General ; Bronchoscopes ; Fiber Optic Technology ; Humans ; Intubation, Intratracheal ; adverse effects ; methods ; Observation ; Surgery, Plastic ; Task Performance and Analysis ; Treatment Outcome

BACKGROUND: The balloon dilatation technique plays an important role in the correction of kyphosis. A balloon catheter can enlarge the spinal cavity in kyphoplasty followed by injection of bone cement under low pressure to lay a foundation for the stability of the spine. OBJECTIVE: To explore the feasibility of balloon dilation with injectable calcium sulfate cement for tibial plateau fractures and to analyze its clinical effect. METHODS: Twenty-four upper tibia specimens of the adults were taken to make the Schatzker Ⅲ collapsed fracture model of the tibial plateau. Then, these specimens were randomized into three groups: the standard group was subjected to poking reduction with autologus bone grafting and screw internal fixation, the bone cement group was inflated with balloon dilatation followed by calcium sulfate cement injection, and the combined group was treated with fixation with cancellous bone screws and balloon dilatation followed by injection of calcium sulfate cement. The general situation of reduction and fixation was observed and the reduction effect was measured. RESULTS AND CONCLUSION: (1) Fixation effect in the model: All three models were well reset, and the average displacement of the standard group, the simple bone cement group and the bone cement screw group was (-0.22±0.62), (-0.23±0.67), and (-0.20±0.69) mm, respectively. There was no significant difference in the displacement between the three groups (P ＞ 0.05). (2) Clinical application: One case of Schatzker type Ⅱ fracture of the left tibial plateau was treated with cancellous bone screw fixation and balloon dilatation followed by injection of calcium sulfate cement. X-ray results showed calcium sulfate cement was visible at 3 postoperative days. At 30 postoperative days, the patient presented with good joint range of motion, and the calcium sulfate was partially absorbed on the X-ray film. At 60 postoperative days, the patient appeared to have no joint extension disorder, and fracture healing and absorption of calcium sulfate as shown by X-rays. To conclude, the balloon dilation with injectable calcium sulfate cement for the treatment of tibial plateau fracture is feasible and has clinical value.

OBJECTIVETo establish a specific procedure for the high-risk screening and diagnosis of organic acidurias and other inherited metabolic diseases in China.

METHODSA nation-wide network for the high-risk screening and diagnosis of genetic metabolic diseases was formed to facilitate the collaboration. Urine samples were collected using filter paper from patients with clinical symptoms suspicious of inherited metabolic diseases. The samples were eluted with distilled water and internal standards were added. Samples were treated with hydroxylamine hydrochloride to form oximes to improve the recoveries of 2-ketoacids. Urinary organic acids were extracted with ethyl acetate and diethyl ether under acidic condition. After dehydration, the combined organic phase was evaporated to dryness with nitrogen. The residues were added with BSTFA + 1%TMCS and heat incubated to form the trimethylsilyl derivatives, and then were analyzed on an Agilent 5890/5973N gas chromatography-mass spectrometer (GC-MS), with a 7683 series auto-sampler. The peaks were identified by reference to a mass spectral library.

RESULTSTotally 352 samples were collected from the network collaborating hospitals since 2001. Thirty-four (9.66%) cases of various inherited metabolic diseases were diagnosed with an age range of 2 days to 14 years. The disease profile was consisted of methylmalonic acidemias (6), alpha-keto-glutaric aciduria (5), tyrosinemia type I (4), dicarboxylic aciduria (4), multiple carboxylase deficiency (3), phenylketonuria (3), lactic acidemia (3), propionic acidemia (2), ornithine transcarbamoylase deficiency (1), ethylmalonic-adipic aciduria (1), glutaric aciduria type II (1) and 3-methylcrotyl CoA carboxylase deficiency (1). The most common clinical symptoms and signs included mental and developmental retardation, convulsion, musculotonic abnormality and jaundice. Routine laboratory tests often revealed metabolic acidosis, hypoglycemia and hyperammonemia, etc.

CONCLUSIONUrine organic acids analysis by GC-MS remains to be the most important technique for the high-risk screening and diagnosis of inherited metabolic diseases. Use of urine filter paper for sample collection and analysis in advanced genetic metabolic centers is a practical approach to extend the diagnostic capacity and improve the management of such diseases in China. Collaborative network played a critical role in the success of the program.

Adolescent ; Carboxylic Acids ; urine ; Child ; Child, Preschool ; China ; Female ; Gas Chromatography-Mass Spectrometry ; methods ; Humans ; Infant ; Infant, Newborn ; Male ; Mass Screening ; methods ; Metabolic Diseases ; diagnosis ; urine

BACKGROUND: Bone tissue engineering based on pluripotent stem cells (PSCs) is a new approach to deal with bone defects. Protocols have been developed to generate osteoblasts from PSCs. However, the low efficiency of this process is still an important issue that needs to be resolved. Many studies have aimed to improve efficiency, but developing accurate methods to determine efficacy is also critical. Studies using pluripotency to estimate efficacy are rare. Telomerase is highly associated with pluripotency. METHODS: We have described a quantitative method to measure telomerase activity, telomeric repeat elongation assay based on quartz crystal microbalance (QCM). To investigate whether this method could be used to determine the efficiency of in vitro osteogenic differentiation based on pluripotency, we measured the pluripotency pattern of cultures through stemness gene expression, proliferation ability and telomerase activity, measured by QCM. RESULTS: We showed that the pluripotency pattern determined by QCM was similar to the patterns of proliferation ability and gene expression, which showed a slight upregulation at the late stages, within the context of the general downregulation tendency during differentiation. Additionally, a comprehensive gene expression pattern covering nearly every stage of differentiation was identified. CONCLUSION: Therefore, this assay may be powerful tools for determining the efficiency of differentiation systems based on pluripotency. In this study, we not only introduce a new method for determining efficiency based on pluripotency, but also provide more information about the characteristics of osteogenic differentiation which help facilitate future development of more efficient protocols.
Bone and Bones ; Down-Regulation ; Gene Expression ; In Vitro Techniques ; Methods ; Mouse Embryonic Stem Cells ; Osteoblasts ; Pluripotent Stem Cells* ; Quartz Crystal Microbalance Techniques ; Telomerase* ; Up-Regulation