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 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

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 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.

OBJECTIVE: To study expression of inducible nitric oxide synthase (iNOS) in oral squamous cell carcinomas (OSCC) and its relations to microvessel density (MVD) and lymphatic metastasis. METHODS: With 9 cases of normal oral mucosa as control, the expression rate of iNOS in 41 cases of OSCC was evaluated by immunohistochemstry SP staining. With CD34 as label, MVDs of these cases were also detected. RESULTS: The iNOS expression rate of OSCC cases was 63.41%(26/41), while no positive expression was seen in 9 cases of normal oral mucosa. Anymore, the expression rate of N+ group was 85.00% while that of N group 42.90%, the rate between them was significantly different (P<0.01). There was correlation between MVD and lymphatic metastasis (rs=0.51, P<0.01). MVDs in groups of different iNOS expression( approximate, equals +++) were 29.667+/-6.945, 34.833+/-4.579, 46.357+/-6.687, 54.167+/-5.565, respectively, and were significantly different with each other (P<0.001). CONCLUSION: There is high expression rate of iNOS in OSCC, and it has close relation to lymphatic metastasis; angiogenesis may facilitate lymphatic metastasis of OSCC, and expression of iNOS has positive relation to MVD in OSCC.

OBJECTIVETo investigate the prevalence of nonalcoholic steatohepatitis (NASH) in obese children and its possible mechanism.

METHODSThree subgroups were classified according to their body mass index (BMI) in 123 obese children with BMI over 23 aged 7 to 16:34 cases with BMI> or =31 group; 57 cases with 25< or =BMI<30 group; 32 cases with 23< or =BMI<25 group. Ultrasonographic and biochemical parameters including serum alanine aminotransferase (ALT), serum aspartate aminotransferase (AST), serum cholesterol, serum triglyceride, serum uric acid and free glucose to free insulin ratio (FGIR) were assayed. Twenty four children suspected as benign acanthosis nigricans underwent skin biopsy and its association with nonalcoholic steatohepatitis was also discussed.

RESULTSNinety-nine children (80.49 %) showed abnormal hepatic sonograms and 54 were diagnosed as NASH with the prevalence of 43.90%. Compared with the other two groups, BMI> or =31 group was significantly higher in prevalence of abnormal hepatic sonograms, NASH, decreased FGIR and risk of benign acanthosis nigricans (P<0.01). Fifty-four children diagnosed as NASH showed significantly higher incidence of hyperlipidemia, insulin resistance and higher body mass index as compared with 24 subjects without fatty liver changes. In 54 NASH children, 20(37.04%) had benign acanthosis nigricans. By bivariate analysis, ALT and AST were correlated well with BMI, cholesterol, triglyceride and FGIR (r(s)=0.413, 0.290, 0.379, -0.477, P<0.01; r(s)=0.359, 0.349, 0.348, -0.369, P<0.01).

CONCLUSIONThere is a high prevalence of nonalcoholic steatohepatitis in simple obese children and high incidence of benign acanthosis nigricans in NASH subjects. BMI> or=30 is a high risk factor of being NASH. Severe disturbance of lipid metabolism and insulin resistance may be involved in the mechanism of NASH.

Acanthosis Nigricans ; etiology ; Adolescent ; Body Mass Index ; Child ; China ; epidemiology ; Fatty Liver ; epidemiology ; etiology ; Female ; Humans ; Insulin Resistance ; Male ; Obesity ; complications ; Prevalence

OBJECTIVETo assess the diagnostic value of the simplified gonadorelin stimulation test for precocious puberty.

METHODSTwo hundred and ninety-two girls with signs of advanced breast development received the gonadorelin stimulation test. According to the result of gonadorelin stimulation test, the girls were divided into 3 groups: 151 with central precocious puberty (CPP),119 with premature thelarche (PT) and 22 with peripheral precocious puberty (PPP).

RESULTSLH or FSH levels at 15 min, 30 min, 60 min in PPP group were not significantly different (P>0.05). Those were significantly different in PT group (P<0.01). The highest levels of LH were at 30 min and the highest levels of FSH were at 60 min. LH or FSH levels at 15 min, 30 min, 60 min in CPP group were significantly different (P<0.01) with the highest levels at 30 min. The ratio of basal LH and FSH >0.2 had a diagnostic sensitivity of 48.3 % and specificity of 69.7%. Taking the LH/FSH ratio >0.9 at 15 min, 30 min, 60 min as cut-off value, the diagnostic sensitivity was 80.1%, 68.9% and 38.4%, and the specificity was 90.8%, 96.6% and 69.7%, respectively.

CONCLUSIONThe LH/FSH ratio>0.9 at 15 min after gonadorelin stimulation test can be used as a cut-off value to differentiate CPP from PT and blood sample at 60 min were not necessary.

Child ; Female ; Follicle Stimulating Hormone ; blood ; Gonadotropin-Releasing Hormone ; Gonadotropins ; blood ; Humans ; Luteinizing Hormone ; blood ; Predictive Value of Tests ; Puberty, Precocious ; diagnosis ; Sensitivity and Specificity

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

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