Objective:To study the effects of Tongxinluo,a traditional Chinese medicine,on the proliferation of peripheral blood-derived endothelial progenitor cells(EPCs)in vitro.Methods:The Tongxinluo solution was prepared through ultrasonication according to the pervious literature.The mononuclear cells(MNCs)were isolated from the peripheral blood with Ficoll by density gradient centrifugation.MNCs were suspended in Medium 199 containing 20% fetal blood serum(FBS)and vascular endothelial growth factor(VEGF).After cultured for 7 d,the attached cells were characterized by Di-LDL uptaking and FITC-lectin binding by laser confocal microscope,and further identified through detection of CD34 and CD133 expression by flow cytometry.Then the cultured EPCs were incubated with Tongxinluo at a series of concentrations(0,50,100,200,500, 750,1000?g/ml)for different durations(0,6,12,24 and 36 h).The cell morphology was observed and cell proliferation was determined by MTT assay.Results:Incubation with different concentrations of Tongxinluo increased the proliferative ability of EPCs.Tongxinluo at 500?g/ml had the most prominent effect on proliferation and the effect increased as time went by and reached peak at 36 h(growth rate 54.18%,P

Objective The purpose of this paper is to screen inborn errors of meta bolism (IEM) by analyzing urinary components, so as to provide laboratory guide for their diagnosis and therapy.Methods Urine samples of patients suspected to have IEM were collec- ted.Urea was de compo sed with urease and n-heptadecanoic acid was added as internal standard.Protein was denatured with ethanol and precipitate was removed by centrifugation,dried b y evaporation, the residue was trimethylsilylly derivatized with BSTFA/TMCS,and then analyzed with GC-MS for quantification of organic acids, amino acids,suga rs, polyols, purines and pyrimidines, simultaneously. This procedure is denom inated as urease pretreatment-gas chromatography-mass spectrometry (UP-GC-MS) internationally.Results Urinary samples of 327 patients from 6 provinces, cities and autonomous regions were analyzed,and 16 kinds of 27 cases of IEM were screened out with a positiv e rate of 8.26%,among which there were 3 cases of hyperphenylalaninemia,3 cases of glyceroluria,3 cases of Leigh syndrome, 2 cases of propionic acidemia, 2 case s of methylmalonic aciduria, 2 cases of von Gierke′s disease, 2 cases of fructo se-1,6-diphosphatase deficiency, 2 cases of fructosuria, 1 cases of multiple car boxylase deficiency, 1 cases of glutaric acidemia typeⅠ, 1 cases of maple sy rup urine disease, 1 cases of hyperglycinemia, 1 cases of 3-aminoisobutyric acid uria,1 cases of adult-onset typeⅡcitrullinemia,1 cases of galactosemia and 1 ca ses of Fanconi′s syndrome.Several IEM patients above had died,but satisfactory therapeutic effects had been achieved in some diseases,in cluding multiple carboxylase deficiency,methylmalonic aciduria and galactosemia. Other patients′ condition remained to be followed up.Conclusion Analysis of urinary components by UP-GC-MS provides a valuable tool for screenin g of IEM and the results will help to provide effective diagnostic and therapeut ic guide for the patients. J Appl Clin Pediatr,2005,20(2):142-144

Objective Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD, OMIM 605814 ) is a novel autosomal recessive disease results from mutations in the gene SLC25A13 that encodes for citrin, a liver-type aspartate/glutamate cartier located in the mitochondrial inner membrane. Most of the Chinese NICCD patients diagnosed by genetic analysis had the sameSLC25A13 mutations as Japanese, however, in some cases, the known mutations were not detected. This research aimed to identify novel SLC25A13 mutations in Chinese NICCD patients and to explore the experimental conditions for their genetic diagnosis.Methods Genomic DNA was extracted from blood samples of 3 NICCD patients from Taiwan (P757), Guangdong (P1194) and Hebei (P1443) Province of China, respectively; and all the 18 exons and their flanking sequences of SLC25A13 gene were sequenced. Furthermore, the identified novel mutations were diagnosed by amplification with PCR, digestion with corresponding restriction endonuclease, and agarose gel electrophoresis.Results Three novel mutations identified in SLC25A13 gene of the 3 NICCD patients were an abnormal splicing IVS7-2A>G (P757), a missense A541D (c. 1622C > A, P1194) and a nonsense R319X (c. 955C > T, P1443). The PCR-RFLP procedures for their genetic diagnosis were also established, with specific fragments on electrophoresis after digestion of the PCR products with three different restriction endonucleases Msp Ⅰ, Hpy188Ⅰ and Taq Ⅰ, respectively.Conclusions The three novel mutations in SLC25A13 gene of Chinese NICCD patients were first identified, suggesting that SLC25A13 mutation distributed in Chinese population is somewhat different from that in Japanese. Moreover, the PCR-RFLP diagnostic procedures established in this research provide valuable tools not only for the genetic diagnosis of NICCD but also for further molecular epidemiologic investigations in Chinese population.Acknowledgement We are grateful to all research subjects and their family members for their cooperation, and to many members of medical staff who contributed much to this research. This study was financially supported partially by Guangdong Provincial Research Grant for Science and Technology (No. 2004B50301008) and the Major State Basic Research Development Program of China (No. 2007CB511901 ), and by Grants-in-Aid for Scientific Research (B: Nos. 16390100 & 19390096 ) and for Asia-Africa Scientific Platform Program (AASPP) from the Japan Society for the Promotion of Science.

OBJECTIVETo investigate the effects of small interfering RNA targeting connective tissue growth factor (CTGF) on rat transforming growth factor beta (TGF beta)/Smads signal pathway.

METHODSChemically synthetic siRNA targeting CTGF was transfected into HSC T6 and then they were injected into rat livers through their intraportal veins. At the same time these rats also received CCl4 subcutaneously every three days for 6 consecutive weeks. Untreated HSC T6 or/and rats with random siRNA treatment served as controls. Total RNA or/and protein in HSC T6 and rat hepatic tissues were extracted. The expressions of CTGF and TGF beta 1, Smad2, 3 and 7 genes were detected by reverse transcription-polymerase chain reaction (RT-PCR) and/or Western blot.

RESULTSCTGF siRNA significantly reduced the expression of CTGF protein in HSC T6. At 48 h after CTGF siRNA treatment, the down-regulation of CTGF protein was the most significant, up to 94%+/-4% (t=46.196, P less than 0.01), but the expressions of TGF beta 1, Smad2, 3 and 7 mRNA showed no differences in HSC T6 compared with the blank controls. Six weeks after CCl4 injections, prominent up-regulations were observed in the gene expressions of CTGF and TGF beta 1 in saline control or siRNA-treated rat livers. Administering CTGF siRNA for six weeks markedly attenuated the induction of CTGF and TGF beta 1 genes; the expressions of CTGF and TGF beta 1 protein decreased by 95%+/-2% (F=21.234, P less than 0.01) and 74%+/-8% (F=13.464, P less than 0.05), respectively, whereas Smad2, 7 protein expressions were not affected.

CONCLUSIONSilencing the CTGF gene can suppress the TGF beta /Smads signal pathway in rat livers.

Animals ; Connective Tissue Growth Factor ; metabolism ; Gene Silencing ; Male ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Smad Proteins ; metabolism ; Transfection ; Transforming Growth Factor beta ; metabolism

Objective: To observe the effect of L-carniti ne (L-CN) in the treatment of congestive heart failure (CHF). Meth ods: Fifty-six cases of chronic CHF randomly received routine treatment (Digitalis, diuretics, vasodilator, ACEI or βblocker) or L-CN (3.0 g/d ,V D×10 d) with routine therapy. Results: The treatment efficiency of L-CN group and control group were 89.3% and 60.7% (P<0.01), respect ively. No adverse reactions related to the drug were observed. Conclusio n: L-CN with routine therapy might be a safe way to the treat CHF.

OBJECTIVETo study the efficacy of bedside treatment by laser photocoagulation for retinopathy of prematurity (ROP) in preterm infants hospitalized in the Neonatal Intensive Care Unit (NICU).

METHODSThe clinical data of 30 cases of ROP who underwent peripheral laser ablation on bedside in the NICU from March to August 2009 were studied retrospectively.

RESULTSA total of 59 eyes from 30 patients received the laser therapy, with a total cure rate of 95%. According to the International Classification of ROP, 26 eyes of 13 infants had zone 1 disease, and 33 eyes of 17 infants had zone 2 disease. The birth gestational age and birth weight as well as corrected gestational age and corrected weight at operation in the zone 1 disease group were significantly lower than those in the zone 2 disease group. The number of laser spots in the zone 1 disease group was significantly higher than that in the zone 2 disease group. The cure rate in the zone 2 disease group (100%) was significantly higher than that in the zone 1 disease group (88%).

CONCLUSIONSLaser retinal photocoagulation on bedside in the NICU is effective for both zone 1 and zone 2 ROP. As compared with the infants with zone 2 disease, the infants with zone 1 disease may have a poor outcome.

Birth Weight ; Female ; Gestational Age ; Humans ; Infant, Newborn ; Laser Coagulation ; Male ; Retinopathy of Prematurity ; surgery

OBJECTIVETo study the genome sequence of hepatitis A virus L-A-1 strain which has been applied for live attenuated vaccine production in China, to compare with other HAV strains, to understand some characteristics of L-A-1 strain, and to find the mechanism of attenuation and cell adaptation.

METHODSGenome fragments were prepared by antigen-capture PCR from infected cell (2BS), PCR products were cloned into T vector, sequenced and analyzed by using bioinformatics program.

RESULTSAnalysis of the genomic sequences(nt 25-7,418) showed that the open reading frame contains 6,675 nucleotides in length encoding 2,225 amino acids. Sequence homology comparison showed 98.00% and 94.00% homology at nucleotide level, and 98.51% and 98.65% homology at amino acid level with international strains MBB and HM 175, respectively. Through comparison with other attenuated, cell adapted and cytopathic effect (CPE) strains, L-A-1 strain had mutation at nt 152, 591, 646, 687 and insertion at nt 180-181 in 5?NTR and had mutation at nt 3,889 (aa 1 052-Val) in 2B region, these mutations and insertion are molecular basis for cell adaptation; mutation at nt 4,185 (aa 1 152-Lys) in 2C region should be attenuated marker; deletion in 3A region (nt 5,020-5,025) that caused two amino acids deletion is virus fast growth basis.

CONCLUSIONThrough analyzing L-A-1 strain genomic sequence, certain sites related to cell adaptation and attenuation were found.

Adaptation, Biological ; genetics ; Amino Acid Sequence ; Base Sequence ; Gene Deletion ; Genome, Viral ; Hepatitis A Vaccines ; genetics ; Hepatitis A virus ; genetics ; growth & development ; Mutation ; Open Reading Frames ; genetics ; Sequence Homology ; Vaccines, Attenuated ; genetics

OBJECTIVETo evaluate the accuracy and related affecting factors of the intra-operative somatosensory evoked potential monitoring in cervical and thoracic surgery.

METHODSCortical somatosensory evoked potential (CSEP) monitoring and sub cortical somatosensory evoked potential (Sub-CSEP) monitoring were performed in cervical and thoracic surgery. Somatosensory evoked potential (SEP) changes were recorded during anaesthesia and operation and postoperative, which could be used to evaluate accuracy of SEP.

RESULTSBilateral CSEP wave abnormalities were related to anaesthesia, decreasing wave amplitudes did not reach the alarming standard. Intra-operative manipulation to affect spinal cord would influence iso-lateral wave abnormality of CSEP and sub-CSEP, decreasing amplitudes reached the alarming standard. Local hypothermia such as cold water irrigating spinal cord would be to prolong the latent period. Low mean arterial pressure (MAP) mostly influenced amplitudes. Changes of SEP in local hypothermia and MAP did not reach the alarming standard.

CONCLUSIONSCSEP and Sub CSEP can reflex physiopathological condition of spinal cord, it is useful in evaluating spinal cord function and providing the safety for cervical and thoracic surgery.

Adult ; Aged ; Anesthesia ; Cervical Vertebrae ; surgery ; Evoked Potentials, Somatosensory ; Female ; Humans ; Intraoperative Complications ; prevention & control ; Male ; Middle Aged ; Monitoring, Intraoperative ; methods ; Reproducibility of Results ; Spinal Cord Injuries ; etiology ; prevention & control ; Thoracic Vertebrae ; surgery

The aim of this study was to explore the changes in cellular senescence related indexes of bone marrow mesenchymal stem cells (BMMSCs) after total body irradiation (TBI). At different time points after 4 Gy irradiation, BMMSCs were isolated from male C57BL/6 mice and cultured. Morphology, senescence-associated beta-galactosidase (SA-beta-gal) staining and cell cycle analysis were used to evaluate the changes in BMMSCs at cellular level while real-time RT-PCR was used to detect the alterations in senescence related gene expression including p16INK4a, p21Cip1/Waf1, p53 and TGF-beta1. The results showed that within 4 weeks after exposure to 4 Gy TBI, the morphology of BMMSCs and the expression level of SA-beta-gal were not significantly changed, the cellular senescence-related cell cycle arrest was not occurred and the senescence related gene expression level was not increased. It is concluded that at the early stage after 4 Gy TBI, the related molecular level of cellular senescence in BMMSCs is not changed.
Animals ; Bone Marrow Cells ; cytology ; radiation effects ; Cells, Cultured ; Cellular Senescence ; radiation effects ; Male ; Mesenchymal Stromal Cells ; cytology ; radiation effects ; Mice ; Mice, Inbred C57BL ; Whole-Body Irradiation

OBJECTIVETo explore the risk factors for irritable bowel syndrome (IBS) among school adolescents in China.

METHODA stratified, randomized study by cluster sampling was conducted, which recruited 51,956 students from high and primary schools in Chinese cities. All students were requested to fill in a questionnaire.

RESULT(1) Factors including class (odds ratio 1.12), excessive intake of pepper (odds ratio 1.17), fried (odds ratio 1.08) and starch-based foods (odds ratio 1.06), gastrointestinal tract infection (odds ratio 2.66), abuse of analgesic (odds ratio 1.49), inheritance (odds ratio 1.83), fatigue (odds ratio 1.32) and repression (odds ratio 1.45) were significantly associated with the presence of IBS (P < 0.05). High protein food (odds ratio 0.90) was a protective factor.

CONCLUSIONDifferent food intake, gastrointestinal tract infection, abuse of analgesic, inheritance and psychological factors might be related to development of IBS in the students of the cities involved in this study.

Adolescent ; Child ; China ; epidemiology ; Humans ; Irritable Bowel Syndrome ; epidemiology ; etiology ; Risk Factors ; Sampling Studies ; Students ; Surveys and Questionnaires