Age Determination by Skeleton ; Body Height ; drug effects ; Child ; Child Development ; drug effects ; Drug Therapy, Combination ; methods ; Female ; Gonadotropin-Releasing Hormone ; administration & dosage ; analogs & derivatives ; therapeutic use ; Growth Hormone ; administration & dosage ; therapeutic use ; Humans ; Menarche ; Puberty, Precocious ; diagnosis ; drug therapy ; physiopathology ; Treatment Outcome

OTX1 gene is one of the pivotal transcriptional factors involved in the neurogenesis.In order to overexpress the OTX1 gene in distinct cell types and find out its contribution to the proliferation and differentiation of stem cells in vitro,OTX1 cDNA was subcloned into lentiviral vectors.The resulting constructions pDUETOTX1,pDUETGFPOTX1 and pDUETGFP were packaged in 293 cells producing viral particles to transduce 293T cells,SY5Y cells,mouse embryonic stem cells and E15 neural stem cells.It was proved that the transferred OTX1 gene was located in the nuclei of the transduced cells in stead of plasma.Lentivirus is an ideal vector delivering gene to different cells.The overexpression of OTX1 in transduced 293T cells were validated by Western blot and immunofluorescence.

Objective To investigate the value and clinical significance of congenital heart diseases (CHD)detection in twins.Methods A total of 1103 twins were included in this study(127 twins were at high risk for CHD).The fetal hearts were scanned by ultrasound using Yagel's heart examination method. Autopsies were done when the pregnancy was terminated.And blood samples from fetal hearts or umbilical veins were used to evaluate fetal chromosomes.A close follow-up was conducted for normal heart cases and another heart examination was done within three months after birth.Results(1)12 twins(1.09%,12/ 1103)had CHD.Among them,4 cases were from the high risk for CHD group(33.3%,4/12),and 8 cases(66.7%,8/12)were from the low risk pregnancy group.(2)Two twins suffered from the same CHD (one pair were both TOF,and the other pair were both rhabdomyoma).One pair of twins had different abnormalities(one baby was TOF,and the other was duodenal atresia with a normal heart).All three pairs of twins chose termination and autopsies were conducted.Unanimous conclusions between prenatal ultrasound and autopsy were obtained.Nine twins were CHD in one baby and a normal heart in the other baby.Seven of them had the same conclusion after delivery.(3)Two twins with CHD were found with fetal abnormal chromosome.(4)1091 cases were not found having any abnormality,however,one fetus from one twin pair was diagnosed with ventricular septal defect(VSD)with abnormal chromosome after birth,and one fetus from another twin pair had patency of ductus arteriosus after birth.(5)The sensitivity of Yagel's heart examination was 82.4% and specificity was 100% in twins.Conclusion Yagel's heart examination is an effective and time-saving method to scan fetal hearts in twins.

In this paper, the RP-HPLC specific chromatography was adopted, with DIKMA-C18 (4.6 mm x 250 mm, 5 µm) as the chromatographic column, with a gradient elution compose of acetonitrile and 0.1% phosphoric acid at flow rate of 0.8 mL · min(-1), the detection wavelength was 220 nm. The difference of the HPLC specific chromatograms between the Lu Dangshen and other different base sources and different producing area of Codonopsis Radix was compared, involved in the similarities and differences of the number and the relative peak area of characteristic peaks in the HPLC specific chromatograms. The HPLC specific chromatograms of Lu Dangshen was established and the relative retention times of seven peaks was determined, and the peaks of codonopyrrolidium B, syringin, lobetyolin, tangshenoside I and atractylenoide III were identified; The HPLC specific chromatograms of Lu Dangshen provided a method for scientific evaluation and effective control the quality of Lu Dangshen from Shanxi famous-region.
Chromatography, High Pressure Liquid ; methods ; Codonopsis ; chemistry ; Drugs, Chinese Herbal ; analysis ; Glucosides ; analysis ; Phenylpropionates ; analysis ; Plant Roots ; chemistry ; Quality Control

OBJECTIVE: To study the correlation of daily living activities with location and severity of trau- matic brain injury (TBI) and to provide a theoretical basis for improving the accuracy of expert opinion. METHODS: Five hundred and one cases of patients with TBI were selected. Detailed records included following: pre-injury situation, location and severity of injury, treatment and education. Daily living activi- ties scale (Barthel index) was applied to test the subjects' daily living activities. The relevance among location and severity of TBI and Barthel index was statistically analyzed. RESULTS: In mild TBI group, there was no significant difference in Barthel index among each location (P>0.05). In moderate TBI group, there were significant differences in Barthel index between subarachnoid hemorrhage and cerebral lobe injury, also between parietal, occipital lobes injury and frontal lobe injury, parietal, occipital lobes injury and temporal lobe (P<0.05), respectively, whereas no significant difference in Barthel index between frontal lobe injury and temporal lobe injury (P>0.05). In severe TBI, there were significant differences in Barthel index between every two different locations (P<0.05). CONCLUSION There is some correlation between the location of TBI and Barthel index, which provides an important reference value for analyzing and determining daily living activities after TBI.
Activities of Daily Living ; Adult ; Brain Injuries/rehabilitation* ; Female ; Humans ; Male ; Outcome Assessment, Health Care ; Trauma Severity Indices

Objective：This study was designed to establish and optimize the research methods for proteome,and to analyze the proteome components of human lung squamous carcinoma cell line NCI H520. Methods： A series of methods, including immobilized pH gradient two dimensional polyacrylamide gel electrophoresis(2DE), silver staining, PDQuest 2DE analysis software, peptide mass fingerprint based on matrix assisted laser desorption/ionization time of flying mass spectrometry (MALDI TOF MS) and SWISS PROT database searching, were used to separate and indentify the proteome of human lung squarmous carcinoma cell line NCI H520. Results： The good 2DE pattern including resolution and reproducibility was obtained. After silver staining, the 2DE image analysis by PDQuest 2DE software had detected average of 1146± 116 spots, and 851± 95 spots were matched. The average matching rate was 73.3％ . There had a good reproducibility of spot position in 2DE map, with average deviation in IEF direction of 1.52± 0.22 mm， while in SDS PAGE direction it was 1.97± 0.13 mm. Sixty spots were incised from silver staining gel randomly and digested in gel by TPCKtrypsin. Fifty four peptide mass fingerprints (PMF) maps were obtained by MALDI TOF MS. The typic peptide masses were searched in the SWISS PROT database by PeptIdent software. Forty four proteins were preliminarily identified. Some of them were cell cycle related proteins such as Cyclin H, some were signal transduction related proteins such as mitogen activated protein kinase, protein kinase C and receptor protein tyrosine kinase ERBB 2, some were oncogene related proteins such as Ras related protein RAB 36, etc. Conclusions： The main proteome research system including IPG 2DE, image analysis, MALDI TOF MS derived PMFs and database searching has been established. The data of NCI H520 obtained by above methods will be useful for the establishment of human lung squamous cell proteome database.

BACKGROUNDThe large phage antibody library is used to obtain high-affinity human antibody, and the Loxp/cre site-specific recombination system is a potential method for constructing a large phage antibody library. In the present study, a phage antibody library vector pDF was reconstructed to construct diabody more quickly and conveniently without injury to homologous recombination and the expression function of the vector and thus to integrate construction of the large phage antibody library with the preparation of diabodies.

METHODSscFv was obtained by overlap polymerase chain reaction (PCR) amplification with the newly designed VL and VH extension primers. loxp511 was flanked by VL and VH and the endonuclease ACC III encoding sequences were introduced on both sides of loxp511. scFv was cloned into the vector pDF to obtain the vector pDscFv. The vector expression function was identified and the feasibility of diabody preparation was evaluated. A large phage antibody library was constructed in pDscFv. Several antigens were used to screen the antibody library and the quality of the antibody library was evaluated.

RESULTSThe phage antibody library expression vector pDscFv was successfully constructed and confirmed to express functional scFv. The large phage antibody library constructed using this vector was of high diversity. Screening of the library on 6 antigens confirmed the generation of specific antibodies to these antigens. Two antibodies were subjected to enzymatic digestion and were prepared into diabody with functional expression.

CONCLUSIONSThe reconstructed vector pDscFv retains its recombination capability and expression function and can be used to construct large phage antibody libraries. It can be used as a convenient and quick method for preparing diabodies after simple enzymatic digestion, which facilitates clinical trials and application of antibody therapy.

Bacteriophages ; genetics ; Digoxin ; immunology ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; Immunoglobulin Fragments ; biosynthesis ; immunology ; Peptide Library

OBJECTIVETo observe effect of opening point method of ziwu liuzhu on myocardial ischemia in the patient of stroke.

METHODSThirty cases of stroke with myocardial ischemia were treated with opening acupoint according to time of the combination of reinforcing mother point and reducing son point of the heart channel in nazi method of ziwu liuzhu, and 30 cases in the control group were treated with acupuncture at Guangming (GB 37). Changes of electrocardiogram before and after treatment were recorded.

RESULTSST-T segment raised significantly 30 min after treatment with ziwu liuzhu needling method as compared with the control group (P < 0.05), the treatment group being better than the control group.

CONCLUSIONExcessive and deficient changes of the qi and blood in the meridians and collaterals are correlated with time rhythm, and ziwu liuzhu needling method can increase clinical therapeutic effect.

Acupuncture Points ; Acupuncture Therapy ; Humans ; Meridians ; Myocardial Ischemia ; Stroke ; therapy

OBJECTIVEMicrocapsule chemoembolism is a promising treatment of tumors. We describe a deep lingual arterial embolization of tongue carcinoma with microcapsuled carboplatinum.

METHODSLingual artery cast specimens from cadavers were microscopically examined, and 78 patients with tongue cancer were recruited and treated with the deep lingual arterial embolization therapy.

RESULTSMicrocapsule embolism occurred approximately at the fifth or sixth level of the deep lingual artery branches. The five-year survival rate was 88.5% (69 out of 78), and the ten-year survival rate 52.6% (41 out of 78).

CONCLUSIONThe deep lingual arterial embolization of tongue carcinoma with microcapsuled carboplatinum is an effective therapy to treat carcinoma in mid-margin or mid-body of the tongue.

Antineoplastic Agents ; administration & dosage ; Capsules ; Carboplatin ; administration & dosage ; Chemoembolization, Therapeutic ; methods ; Drug Carriers ; administration & dosage ; Humans ; Injections, Intra-Arterial ; Tongue ; drug effects ; Tongue Neoplasms ; therapy

In vitro gene delivery, polyethyleneimine (PEI) has been described as one of the most efficient nonviral vector. Herein the formation mechanism of PEI/DNA complexes is elucidated. The transition phase of "bead-on-string" structure in the formation of complexes was supposed to exist through spectroscopy, electrophoresis and transmission electron microscopy (TEM) technology. The construction of PEI/DNA complexes is related closely to the characteristics of PEI and DNA plasmid. As well as the dominant electrostatic effects, the nonelectrostatic interactions were thought to be partially responsible for the presence of PEI/DNA complexes even in the high ionic strength. The surface charge of complexes particles increased with the N/P ratio, but the absolute value of zeta potential was lower at the N/P ratio of 8 and 12, perhaps attributed to the use of larger DNA plasmid. As a result, the repulsion between particles was decreased and prone to aggregate to the structure like a clustered grape-string in the solution. Interestingly, contrast to the formation behavior of complexes, the PEI/DNA complexes aggregated primarily due to hydrophobic interactions while electrostatic attractions play a little role in the complexes particles aggregation in different concentrations of salt solutions. Comparable transfection efficiency in HepG2 cells was observed for the Lipofectamine 2000 and PEI/DNA complexes at the N/P ratio of 12, and showed that larger or aggregable complexes could transfect the cells in some different mechanisms.
DNA ; genetics ; Drug Carriers ; Gene Transfer Techniques ; Genetic Therapy ; Polyethyleneimine ; chemistry ; Transfection