BACKGROUND: Myogenetic seed cells are the fundamental for construction of tissue-engineered smooth muscle, and optimizing the amplification of seed cells is the key in further clinical application. OBJECTIVE: To analyze the myogenetic potential of rabbit adipose-derived stromal cells induced with the combination of MyoD, transforming growth factor β1 and 5-azacytidine, and to investigate a new way to induce cells. METHODS: The rabbit abdominal fats were isolated and then the adipose-derived stromal cells were separated by col agenase digestion method for myogenetic induction with different methods: 5-azacytidine induction group, MyoD+transforming growth factor β1 induction group, 5-azacytidine+MyoD+transforming growth factor β1 combination induction group. The blank control group was set. The morphological characteristics of cells were observed at 1, 4, 8, 12, 16, 20, 24 and 28 days after induction, and the col agen type Ⅲ level were detected with 4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry and immunohistochemistry. RESULTS AND CONCLUSION: Compared with the other groups, the cel activity was higher in the combination induction group and reached peak at 16 days after induction, the number and volume of myotubes were increased at 20 days with regular arrangement, and desmin showed strong expression. Cel cycle showed the ratio of adipose-derived stromal cells in the DNA replication phase was increased in the combination induction group, the ratio of cells in the clearance period was decreased; the level of col agen type Ⅲ was increased significantly, and the difference was significant. The results indicate that 5-azacytidine combined with multiple factors can promote the differentiation of adipose-derived stromal cells into myoblasts with significant cel proliferation, which is considered as the ideal method to in vitro induction of myoblasts.

The migration-time in transient isotachophoresis (tITP) separation is affected by sample salinity as the dependence of ITP time on sample-zone conductivity.The sample-to-sample variation of migration-time in microchip tITP-CGE analysis is an undesired factor for precise DNA sizing.In this work, a DNA sizing method that based on relative migration-time proportion (RMP) was developed to eliminate the effect of sample salinity on sizing precision.RMP was defined as the ratio of the migration-time difference between the target fragment and the lower marker to that between the upper marker and the lower marker.The RMP values were tested to be reproducible in microchip tITP-CGE separations irrespective of sample salinity.Size of a target DNA was predicted by fitting its RMP value to the equation derived from RMPs of standard DNA ladder vs.DNA sizes.The precision and reproducibility of the sizing method were validated testing multiple standard PCR amplicons.Experimental results showed that the RMP method is simple and reliable, thus well suited to precise DNA sizing with microchip tITP-CGE analysis.

Objective To evaluate the biological potential of surface topography of biomimetic matrix combined PRP gels with RGD modification. Methods Surface topography of nβ-TCP/Cs/PCL matrix was made by Nd: YAG laser, tissue-engineered bone was constructed in the following ways: ADSCs were loaded to nβ-TCP/Cs/PCL matrix with PRP gels plus RGD modification (group A), ADSCs were cultured to nβ-TCP/Cs/PCL matrix with RGD modification (group B), ADSCs implantation with topography-surfaced treatment of matrix (group C), ADSCs cultivation with smooth-surfaced treatment of matrix (group D). SEM and CLM were used to observe cellular pattern, survival rate, cell activity, ALP and collagen type Ⅰ level were detected at 1, 4, 8, 12, 16, 20, 24,28 days. Runx2 and OPG expressions were assayed at different interval. Results Under observation of SEM and CLM, new tissue showed more remarkable cell proliferation with abundant ECM in group A. Compared with other groups, the survival rate in group A was significantly higher (88.16 ± 1.29, P ＜ 0.05),and the level of cell activity, ALP, collagen type Ⅰ were significantly higher (0.92± 0.13, 87.27 ± 3.08, 93.27 ± 3.91, P＜ 0.05), and remarkable expression of Runx2 and OPG was also seen. Conclusion Topography-surfaced treatment of matrix combined PRP gels with RGD modification enhances the cell proliferation and acts as a feasible osteopromotive method.

Objective To develop a phenylketonuria (PKU) screening method based on a compact disk (CD) type microfluidic chip capable of generating reciprocating flow within the microchannels that facilitate rapid DNA hybridization. Methods This microfluidic device consists of a two-layer structure: a polydimethylsiloxane (PDMS) top layer containing 12 DNA hybridization microchannels, and a bottom glass layer with immobilized hydrogel conjugated DNA arrays. The DNA arrays included R243Q, V245V and the blank control probes. When the CD device was spun, the PCR products were driven into the hybridization channel by centrifugal force. When the rotation of the CD device was stopped, capillary force pulled the PCR products solution to flow back to the channel. After the on-chip hybridization, the hybridization signals were captured on a fluorescence microscope. The specificity, detection limitation and reproducibility of this device were evaluated. Thirty DNA samples from pregnant women with suspected PKU were detected by this device.Then the results were compared with DNA sequencing results. Results With the compact disk type microfluidic chip, the hybridization time could be reduced to 15 min, sample consume could be as low as 1. 5 μl and the detection limitation was 0. 7 ng/μl. With the chip based method, samples of PKU patients and healthy controls were detected and the results were consistent with DNA sequencing results. Five different batches of chips and five micro-channels of each chip were selected to test one PKU patients with V245V mutation. All the results were positive, indicating good reproducibility. Four cases of V245V mutation and 1 case of R243Q mutation were found in 30 suspected PKU carried pregnant women. Conclusion The compact disk microfluidic device has advantages of simple, rapid and highly sensitive, thus is well suited to PKU screening.

BACKGROUND: There are less researches on the relationship of parents'feeding behavior with nourishing and developing status of infants.OBJECTIVE: To study the relationship between the parents' feeding behavior and the nourishing and developing status of infants.DESIGN: A cross-sectional study.SETTING: Center of Maternal and Children' s Health,the First Hospital of Xi'an Jiaotong University.PARTICIPANTS: The study was carried out during January to December in 2002,at the Center of Maternal and Children' s Health,The First Hospital of Xi'an Jiaotong University. A total of 346 infants,aged 0 to 18 months old,were selected from the Fuping county and Chengcheng county of Weinan area by using the stratified duster sampline method. Inclusive criteria: infants with the gestation period between 37 to 42 weeks,without obvious diseases,no congenital abnormality; Exclusive criteria: infants with congenital disease or abnormality. Totally 324 infants,who were in accordance with the above criteria,were divided into 5 groups according to their month age.METHODS: The nursing persons of the infants were called at their houses,and the self-designed "Investigation Tables on Infant's Nutrition"were filled out.MAIN OUTCOME MEASURES: ① Status of slight malnutrition of infants of different month ages; ② Status of nutrition and development of infants,within 4 months, with different feeding behaviors; ③ Effects of beginning time,quality and frequency of complementary food on the nourishing and developing status of infants.RESULTS: In the first 4 months after birth,the rate of breast-feeding was high and the nourishing status of the infants was good. But the nourishing and developing status of infants became comparatively worse after 4 months age,because of inappropriate complementary food. As compared the slight malnutrition infants with normal infants,there were significant differences in the beginning time and kinds of complementary food(P<0.01).CONCLUSION: There is close relationship of the feeding behaviors with the nourishing and developing status of the infants.

BACKGROUNDTo provide a simple, specific and early serodiagnostic technique for the patients with aseptic meningitis caused by echovirus.

METHODSAn indirect enzyme-linked immunosorbent assay (ELISA) has been developed to detect echovirus-IgM and the specificity and availability of the assay were also examined.

RESULTSIn 78 cerebrospinal fluid (CSF) specimens which came from the children with aseptic meningitis, the positive rate was 17.9(14/78). In 64 CSF collected from non-aseptic meningitis (bacterial meningitis and cerebral trauma), the positive rate was 1.56(1/64). In 5 CSF specimens which were ELISA positive, the positive rate of neutralization test (NT) was 4/5, all the specimens which were ELISA negative were NT negative. In this assay there was no cross-reaction with poliovirus, Coxsackie virus B type 1-6 and A type 7. By blocking and destructive test of specific IgM, all CSF specimens with ELISA positive became negative.

CONCLUSIONSThe established indirect ELISA was specific and reliable. The te st was quick, simple and available, which is suitable for early and specific clinical diagnosis, and will be greatly significant to clinical treatment.

Adult ; Antibodies, Viral ; cerebrospinal fluid ; Child ; Child, Preschool ; Echovirus Infections ; diagnosis ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunoglobulin M ; cerebrospinal fluid ; Infant ; Infant, Newborn ; Meningitis, Aseptic ; diagnosis ; virology

BACKGROUNDTo clone the type common antigen gD of human herpes simplex virus I, II (HSV-I, II), the authors constructed recombinant expression vector Pmal-c2/gD and induced to express the fusion protein MBP-gD.

METHODSThe authors extracted HSV DNA,amplified gD gene by PCR assay and directly cloned it into prokaryotic expression vector pMAL-c2, then transformed it into E.coli DH5alpha. After proved to be correct by PCR, double enzyme digestion and sequencing, the fusion protein is induced to express by IPTG and detected by both Western blot and ELISA.

RESULTSThe constructed expression vector pMAL-c2/gD can be expressed with high efficiency. The product expressed was about 35.5% of the total bacterium proteins by SDS?PAGE analysis and was found nearly 39% as soluble protein,61% as inclusion in cytoplasm.

CONCLUSIONSThe authors constructed recombinant expression vector pMAL-c2/gD, the Western blotting result showed that the recombinant protein could be identified with gD specific monoclonal antibody DL6. Therefore the protein was of natural antigenic structure of gD.

Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Humans ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; Viral Envelope Proteins ; biosynthesis ; genetics

Based on a discussion on PACS and the way its image workstation obtains scanned sequential images, this paper presented a method of 3D surface construction and visualization on PACS workstation. Guest/Server structure was used between PACS application entities. Image storing and transmission were realized by service classes established by DICOM standards. Relation database was used to arrange the stored sequential images. Image workstation transformed the sequential images obtained from PACS net into volume data field. 3D reconstruction and rendering results were obtained by using surface-rendering and volume-rendering methods, which made the 3D construction results acquire vivid 3D structure details of high fidelity and strong sense of reality. 3 sets of application results were also presented in this paper.
Humans ; Image Processing, Computer-Assisted ; methods ; Imaging, Three-Dimensional ; Magnetic Resonance Imaging ; Radiology Information Systems ; Tomography, X-Ray Computed ; User-Computer Interface

It is an important morphological research method to reconstruct the 3D imaging from serial section tissue images. Registration of serial images is a key step to 3D reconstruction. Firstly, an introduction to the segmentation-counting registration algorithm is presented, which is based on the joint histogram. After thresholding of the two images to be registered, the criterion function is defined as counting in a specific region of the joint histogram, which greatly speeds up the alignment process. Then, the method is used to conduct the serial tissue image matching task, and lies a solid foundation for 3D rendering. Finally, preliminary surface rendering results are presented.
Algorithms ; Image Processing, Computer-Assisted ; methods ; Imaging, Three-Dimensional ; Microtomy ; methods

Objective To study the clinical use of indocyanine green (ICG) fluorescence imaging in laparoscopic liver surgery.Methods The clinical and pathological data of 68 patients who underwent laparoscopic hepatectomy using the ICG fluorescence imaging technique during the study period from September 2016 to October 2018 in Zhongnan Hospital,Wuhan University were retrospectively analyzed.Analysis was carried out on the surgical methods,fluorescence navigation methods,ICG injection time and dose,tumor characteristics,and pathological studies of the resected specimens.Results Of 68 patients,3 patients were converted to open surgery,and the remaining 65 patients completed the ICG fluorescence laparoscopic hepatectomy.Thirty-two of these 65 patients underwent ICG fluorescent guided laparoscopic anatomical resection of lower hepatic segment / hepatic hemilivers (positive staining in 17 patients,negative staining in 15 patients),with 19 patients successfully staining with ICG(19 / 32,59.4％).Postoperative histopathology showed primary hepatic solid tumors (n=31),secondary liver tumors (n=12),hepatic cysts (n=4),hepatic hemangiomas (n =5),hepatolithiasis (n =12) and hepatic focal nodular hyperplasia (n =1).These lesions were combined with hepatitis B liver fibrosis in 29 patients.Conclusions ICG fluorescence imaging positively impacted on laparoscopic liver surgery.Proper preoperative ICG injection was helpful for the identification,localization and intraoperative surgical guidance of tumors,especially for patients with deep-seated and central tumors.As a consequence,oncological and surgical safety of laparoscopic liver surgery was improved.Targeted visualization of liver segments and surgical navigation using intraoperative ICG injections facilitated accurate and precise resection of anatomical liver segments or hemi-hepatectomies.The use of intraoperative ICG fluorescence technology for hepatic hemangioma,hepatic cyst,intrahepatic bile duct stones and other benign liver lesions,helped to improve safety of surgery.