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Biomedical subjects

Yong Liu

Publications and source records attributed to Yong Liu.

At least 19 recordsLinked to original sources

Intramuscular patient-derived xenografts achieve high engraftment rates in gastric cancer: implications for pharmacodynamic testing and genomic biomarker discovery.

BACKGROUND: Gastric cancer (GC) exhibits marked inter-patient heterogeneity, limiting empirical chemotherapy efficacy. Patient-derived xenograft (PDX) models preserve the molecular features of parental tumors and can serve as pharmacodynamic surrogates, but conventional subcutaneous PDX suffers from low engraftment rates. This study evaluated an optimized intramuscular PDX platform for individualized drug testing in GC and applied whole exome sequencing (WES) for biomarker identification (Clinical trial registry: ChiCTR-OOC-17012731). MATERIALS AND METHODS: Ninety-eight treatment-naive GC patients were enrolled between April 2018 and December 2020. Fresh tumor tissues were engrafted into NCG mice by intramuscular transplantation. Drug efficacy was evaluated using tumor cell necrosis rate and Ki-67 expression. WES was performed on 32 engrafted tumorgrafts to characterize driver mutations in fast- and slow-growing subgroups. RESULTS: An engraftment rate of 71.7% (43/60) was achieved, substantially exceeding rates reported in prior studies. Clinical characteristics were independent of engraftment success and outgrowth time (all p > 0.05). Fast- and slow-growing tumorgrafts diverged in frequently altered genes: KMT2C, APOB, CDK12 and MSH2 predominated in fast-growing grafts, whereas TP53, CHD3 and TET2 were enriched in slow-growing grafts. Slow-growing tumorgrafts correlated with longer progression-free survival (p = 0.02). PDX-guided treatment was associated with improved prognosis. CONCLUSIONS: Intramuscular transplantation into NCG mice yields high engraftment rates for GC PDX. PDX-guided chemotherapy selection is associated with favorable outcomes. Driver mutation divergence between fast- and slow-growing tumorgrafts provides candidate prognostic biomarkers.

Animals↗

Long-term heat exposure reshapes muscle molecular regulation and enhances thermal tolerance in Clarias fuscus.

Rapid fluctuations in water temperature driven by global warming have become a major abiotic stressor affecting muscle function in teleost fish. This study examined the effects of long-term thermal conditions on heat tolerance in Clarias fuscus. Fish were maintained for 90 days at either a normal temperature group (NT, 26 °C) or a high-temperature group (HT, 34 °C). Subsequently, muscle histology, and transcriptomic profiles were observed following acute high-temperature exposure (34 °C) and after temperature recovery (26 °C). Histological analysis showed that fish from the NT under acute high-temperature stress exhibited severe muscle damage (atrophy, myofilament disruption, and myolysis), whereas fish from the HT displayed markedly reduced lesions. RNA-seq profiling revealed 5769 differentially expressed genes (DEGs) in the NT and 3292 DEGs in the HT following acute temperature challenges. Functional enrichment indicated that, in the HT, modulation of key cell cycle regulators (e.g., ccna, ccnb, cdk1, cdk2) contributed to alleviating muscle damage caused by temperature fluctuations. In the NT, genes associated with ribosome biogenesis (e.g., nop56, riok2, riok1) were up-regulated and then down-regulated during temperature fluctuation, whereas p53 in the cell cycle pathway showed the opposite expression pattern. These findings demonstrate long-term heat exposure reshapes molecular expression and regulatory mechanisms in the muscle of C. fuscus, thereby enhancing thermal tolerance and adaptability, and providing a theoretical basis for breeding heat-resistant, high-quality aquaculture strains.

Animals↗

Malonate promotes CD8+ T cell memory formation via protein malonylation.

Protein malonylation represents a recently identified posttranslational modification whose role in CD8+ T cell differentiation and functionality remains incompletely understood. In this study, we demonstrate that enhancing protein malonylation through sodium malonate (SM) treatment promotes CD8+ T cell memory formation in response to bacterial infection, subsequently potentiating recall responses. Comparative metabolomic analysis between SM-treated and control CD8+ T cells revealed significant metabolic alterations associated with protein malonylation. We present the first comprehensive proteomic analysis of lysine malonylation in murine CD8+ T cells, identifying 77 malonylation sites across 64 proteins involved in diverse cellular processes, particularly metabolic pathways. Malonylation of STAT6 was confirmed via the use of a specific chemical probe. Notably, we established that malonylation at the lysine 374 site of STAT6 results in increased TCF1 expression, due to alleviated transcriptional repression of TCF1 by STAT6. Collectively, our findings provide compelling evidence that protein malonylation plays a significant role in regulating CD8+ T cell memory formation.

Animals↗

CCR7, CCR8, CCR9 and CCR10 in the mouse hippocampal CA1 area and the dentate gyrus during and after pilocarpine-induced status epilepticus.

The present study showed CCR7, CCR8, CCR9 and CCR10 in the normal Swiss mouse hippocampus at both protein and mRNA levels. CCR7, CCR9 and CCR10 were mainly localized in hippocampal principal cells and some interneurons. CCR9 was also found in the mossy fibres and/or terminals, suggesting an axonal or presynaptic localization, and CCR10 in apical dendrites of pyramidal neurons in the CA1 area. CCR8 was observed in interneurons. Double-labelling immunocytochemistry revealed that most of calbindin (CB)-, calretinin (CR)- and parvalbumin (PV)-immunopositive neurons expressed CCR7-10, except CR-immunopositive cells in which only 10 to 12% expressed CCR8. During and after pilocarpine-induced status epilepticus, progressive changes of each of CCR7, CCR8, CCR9 and CCR10 proteins occurred in different patterns at various time points. Sensitive real-time PCR showed similar change patterns at mRNA level. At the chronic stage, i.e. at 2 months after pilocarpine-induced status epilepticus, significant reduction of CCR7-10 expression in CB-, CR- and PV-immunpositive interneurons may suggest the phenotype change of surviving interneurons. Double labelling of CCR7, CCR8 and CCR9 with glial fibrillary acidic protein (GFAP) at the chronic stage may suggest an induced expression in reactive astrocytes. The present study may, therefore, for the first time, provide evidence that CCR7-10 may be involved in normal hippocampal activity. The demonstration of the progressive changes of CCR7-10 during and after status epilepticus may open a new area to reveal the mechanism of neuronal loss after status epilepticus and of epileptogenesis.

Animals↗

Application of multivariate statistical methods to water quality assessment of the watercourses in Northwestern New Territories, Hong Kong.

Multivariate statistical methods, i.e., cluster analysis (CA) and discriminant analysis (DA), were used to assess temporal and spatial variations in the water quality of the watercourses in the Northwestern New Territories, Hong Kong, over a period of five years (2000-2004) using 23 parameters at 23 different sites (31,740 observations). Hierarchical CA grouped the 12 months into two periods (the first and second periods) and classified the 23 monitoring sites into three groups (group A, group B, and group C) based on similarities of water quality characteristics. DA provided better results with great discriminatory ability for both temporal and spatial analysis. DA also provided an important data reduction because it only used six parameters (pH, temperature, five-day biochemical oxygen demand, fecal coliforms, Fe, and Ni) for temporal analysis, affording about 84% correct assignations, and seven parameters (pH, ammonia-nitrogen, nitrate nitrogen, fecal coliforms, Fe, Ni, and Zn) for spatial analysis, affording more than 90% correct assignations. Therefore, DA allowed a reduction in the dimensionality of the large data set and indicated a few significant parameters that were responsible for most of the variations in water quality. Thus, this study demonstrated that the multivariate statistical methods are useful for interpreting complex data sets in the analysis of temporal and spatial variations in water quality and the optimization of regional water quality monitoring network.

Discriminant Analysis↗

Characterization of red mud derived from a combined Bayer Process and bauxite calcination method.

Red mud can be derived from the processing of bauxite using different methods. The chemical and mineralogical composition of the combined Bayer Process and bauxite calcination red mud (BPBCRM) differs markedly from those of the pure Bayer Process red mud (PBPRM). In this study, red mud derived from a combined Bayer Process and bauxite calcination method was characterized. The results show that pH of the red mud decreased with increasing duration of storage time. Na dominated among the soluble cations, but the concentration of soluble Na decreased with increasing duration of storage time as a result of leaching. Cation exchange capacity also decreased with increasing duration of storage time, probably due to a decrease in pH causing a reduction in negatively charged sites on the red mud particles. Ca was the predominant exchangeable cation in the fresh red mud but the concentration of exchangeable Ca markedly decreased in the old red mud, which was dominated by exchangeable Na. The degree of crystallization and thermal stability of the red mud increased with increasing duration of storage. The acid neutralizing capacity of red mud obtained from this study was about 10 mol kg(-1), which is much greater than the reported values for the pure Bayer Process red mud. Column filtering experiment indicates that the red mud also had a very strong capacity to remove Cu, Zn and Cd from the filtering solution. It is conservatively estimated that the simultaneous removal rates of Cd, Zn and Cu by red mud are over 22,250 mg kg(-1), 22,500 mg kg(-1) and 25,000 mg kg(-1), respectively. The affinity of these metals to the red mud was in the following decreasing order: Cu>Zn>Cd. In general, the fresh red mud retained more heavy metals than the old red mud did.

Aluminum Oxide↗

Preparation of mica-based glass-ceramics with needle-like fluorapatite.

OBJECTIVES: The aim of this study was to prepare mica-based glass-ceramics containing needle-like fluorapatites, and to understand the relationship between the composition, the microstructure and mechanical properties. METHODS: The specimens were prepared by casting and subsequent heat treatment. The crystalline phases in the specimens were determined by X-ray diffraction (XRD). The microstructures of the fractured surface were examined by using a scanning electron microscope with an energy dispersive spectrometer (SEM/EDS). The Vickers hardness and fracture toughness were determined by the indentation method. Statistical analysis of the results was performed by one-way analysis of variance (ANOVA) and Tukey's test. The bioactivity of the glass-ceramics was evaluated by soaking the polished specimens in the simulated body fluid (SBF). RESULTS: Needle-like fluorapatite crystals were successfully obtained in mica-based glass-ceramics by controlling the heat treatment process. The formation of needle-like fluorapatite crystals, instead of particle-like crystals reported in previous studies, can be attributed to the one-dimensional rapid growth of fluorapatite along the c-axis. The mechanical properties of the glass-ceramics are related to the fluorapatite content. The higher the fluorapatite content, the higher the Vickers hardness and the fracture toughness (p<0.01). The bioactivity test showed that the needle-like fluorapatite-containing glass-ceramics possess very good bioactivity. SIGNIFICANCE: As the needle-like fluorapatite crystal has the same morphology as human bones, it endows the mica-based glass-ceramics with good bioactivity. Moreover, the needle-like fluorapatite crystal toughens the base glass-ceramics. Therefore, the mica-based glass-ceramics with needle-like fluorapatite have the potential for the restoration of bone defects and implants.

Aluminum Silicates↗

Sequential priming and boosting with heterologous HIV immunogens predominantly stimulated T cell immunity against conserved epitopes.

BACKGROUND: The effort to develop an effective preventive vaccine against HIV-1 infection is challenged by the wide genetic diversity of HIV-1 among different isolates. OBJECTIVES: To explore a new vaccination strategy by using heterologous HIV immunogens derived from different clades for sequential priming and boosting. METHODS: HIV Env and Gag immunogens derived from Thailand B (B'), C/B' recombinant and A/E recombinant were selected as these three clades account for 29%, 45% and 15% of HIV-1 prevalence in China, respectively. Three humanized fusion genes of env and gag derived from those three clades were synthesized and inserted into DNA and recombinant Tiantan vaccinia vectors as model vaccines. C57BL/6 and Balb/c mice were used as animal model. Peptides spanning the entire Env and Gag were used as stimuli and Elispot assay was used to assess the T cell immunity. RESULTS: Sequential priming and boosting was observed with heterologous HIV immunogens predominantly stimulated T cell immunity against conserved epitopes, whereas a single vaccine derived from one clade or the mixture of multiple vaccines from different clades primarily raised T cells against less conservative or non-conservative epitopes. CONCLUSIONS: This is the first demonstration of a practical strategy to raise immune responses against conserved epitopes. This strategy has important implications for vaccine development against HIV and other pathogens that have high genetic diversity, such as influenza.

AIDS Vaccines↗

Effect of salt on purification of plasmid DNA using size-exclusion chromatography.

In the present study, we compared the performances of size-exclusion chromatography for the purification of plasmid DNA when different concentrations (0.5M, 1M, 2M, respectively) of two types of salt (NaCl and (NH(4))(2)SO(4)) are present in running buffers. Our experiment results displayed that it is not only the resolution of RNA but also those of supercoiled plasmid DNA and host's genomic DNA were increased greatly in the presence of high concentration of water-structure salt. We deduce that two separation modes may be involved in the process: The supercoiled plasmid DNA is influenced mainly by compaction effect and eluted in the size-exclusion mode; whereas, RNA and genomic DNA are influenced mainly by hydrophobic effect due to their stretched and loose structures and eluted in the interaction mode. This method led to an improved efficiency of size-exclusion chromatography.

Chromatography, Gel↗

[Distribution and expression of collagen type II, transforming growth factor beta1 and basic fibroblast growth factor in articular process cartilages of scoliosis].

OBJECTIVE: To look into the character of the expression of collagen type II and transforming growth factor beta1 (TGF-beta1), basic fibroblast growth factor (bFGF) in the apical articular process cartilages of adolescent idiopathic scoliosis (AIS) and congenital scoliosis (CS) patients. METHODS: The articular processes of 22 AIS and 18 CS were collected. The techniques of HE staining, immunohistochemistry and in situ hybridization were adopted in this research. By comparing the apical processes with the end processes, the convex processes with the concave processes, the AIS processes with CS processes, the pathological changes of the articular process cartilages of these patients and the distribution of collagen type II and TGF-beta1, bFGF in them were studied. The images of immunohistochemistry and in situ hybridization were input into the image analysis system and were analyzed semi-quantitatively. The SAS software (8.01) was adopted, and P < 0.05 was defined as the significant level. RESULTS: The expression of collagen type II and TGF-beta1, bFGF in AIS was similar to CS: the concave sides of apexes were higher than the convex sides. The comparisons had statistical significance. There was no statistical significance between upper and lower end vertebrae in convex and concave sides, between convex and concave sides in upper and lower end vertebrae. The apical vertebrae were significantly higher than the ipsilateral sides of upper or lower end vertebrae for collagen type II. There was no statistical difference of the expression at the concave, convex, upper, lower end vertebrae between AIS and CS. CONCLUSIONS: The cartilages of the apical processes show some signs of regression and hypoplasia in scoliosis. The concave side is more severe than the convex side. Increase of collagen type II and TGF-beta1, bFGF in the concave sides of apical processes in scoliosis may be the results of reconstruction of extracellular matrix and the compensation reactions which are caused by abnormal biomechanical forces such as compressive stresses. Compressive stress on the concave sides has more influences on the expression of collagen type II than tensile stress on the convex sides.

Adolescent↗

Nuclear accumulation and activation of nuclear factor kappaB after split-dose irradiation in LS174T cells.

Although radiation-induced gene expression has been extensively studied, most of the studies to date have focused on that after single-dose irradiation. As split-dose irradiation, rather than single-dose irradiation, is usual in clinical situations, we investigated the effects of split-dose irradiation on nuclear factor kappaB (NF-kappaB) in the human rectum carcinoma cell line, LS174T. After either single- or split-dose irradiation with a different interval, nuclear localization of NF-kappaB was examined by Western blot and immunofluorescence and its DNA-binding activity was measured by ELISA-based assay. Irradiation-induced NF-kappaB nuclear accumulation and DNA binding activity increased in a dose-dependent manner. The peak of NF-kappaB nuclear accumulation and DNA binding activity was seen 2 to 6 hours after a single dose of 4 Gy irradiation and returned to control levels after 12 hours. In split-dose irradiation, NF-kappaB activity was similar after the first and second doses of 4 Gy irradiation separated by 12 hours. In addition, NF-kappaB activity was decreased by lengthening the interval between irradiation. The cell survival, which was assessed by colony formation assay, showed inverse correlation to this: the surviving fraction was higher after split-dose irradiation than after single-dose irradiation of the same total dose and it increased as the interval between irradiation was lengthened. Thus the present results showed a correlation between NF-kappaB activation and the repair of sublethal damage in split-dose irradiation.

Cell Line, Tumor↗

Effects of ketamine-midazolam anesthesia on the expression of NMDA and AMPA receptor subunit in the peri-infarction of rat brain.

BACKGROUND: Activation of N-methyl-D-aspartate (NMDA) receptors and alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) receptors play an important role in the neurons death induced by ischemia. The mitigating effect of intravenous anesthetics on ischemic neuron injury is related to their influence on NMDA receptors. This study was performed to investigate the effect of ketamine-midazolam anesthesia on the NMDA and AMPA receptor subunits expression in the peri-infarction of ischemic rat brain and explore its potential mechanism of neuroprotection. METHODS: Thirty Sprague Dawley (SD) rats were subjected to permanent middle cerebral artery occlusion under ketamine/atropine (100/0.05 mg/kg) or ketamine-midazolam/atropine (60/50/0.05 mg/kg) intraperitoneal anesthesia (n=15 each). Twenty-four hours after ischemia, five rats in each group were killed by injecting the above dosage of ketamine or ketamine-midazolam intraperitoneally and infarct size was measured. Twenty-four and 72 hours after ischemia, four rats in each group were killed by injecting the above dosage of ketamine or ketamine-midazolam intraperitoneally. After staining the brain tissue slices with toluidine blue, the survived neurons in the peri-infarction were observed. Also, the expression level of NMDA receptors 1 (NR1), NMDA receptors 2A (NR2A), NMDA receptors 2B (NR2B) and AMPA (GluR1 subunit) were determined by grayscale analysis in immunohistochemical stained slices. RESULTS: Compared with ketamine anesthesia, ketamine-midazolam anesthesia produced not only smaller infarct size [(24.1+/-4.6)% vs (38.4+/-4.2)%, P<0.05], but also higher neuron density (24 hours: 846+/-16 vs 756+/-24, P<0.05; 72 hours: 882+/-22 vs 785+/-18, P<0.05) and lower NR2A (24 hours: 123.0+/-4.9 vs 95.0+/-2.5, P<0.05; 72 hours: 77.8+/-4.1 vs 54.2+/-3.9, P<0.05) and NR2B (24 hours: 98.5+/-2.7 vs 76.3+/-2.4, P<0.05; 72 hours: 67.2 +/-7.5 vs 22.2+/-2.6, P<0.05) expression level in the peri-infarction following ischemia. CONCLUSION: The protective effects of ketamine-midazolam anesthesia on ischemic brain injury may related to decreasing NR2A and NR2B expression.

Anesthetics, Dissociative↗

Boronic acids as inhibitors of steroid sulfatase.

Steroid sulfatase (STS) catalyzes the hydrolysis of steroidal sulfates such as estrone sulfate (ES1) to the corresponding steroids and inorganic sulfate. STS is considered to be a potential target for the development of therapeutics for the treatment of steroid-dependent cancers. Two steroidal and two coumarin- and chromenone-based boronic acids were synthesized and examined as inhibitors of purified STS. The boronic acid analog of estrone sulfate bearing a boronic acid moiety at the 3-position in place of the sulfate group was a good competitive STS inhibitor with a K(i) of 2.8microM at pH 7.0 and 6.8microM at pH 8.8. The inhibition was reversible and kinetic properties corresponding to the mechanism for slow-binding inhibitors were not observed. An estradiol derivative bearing a boronic acid group at the 3-position and a benzyl group at the 17-position was a potent reversible, non-competitive STS inhibitor with a K(i) of 250nM. However, its 3-OH analog, a known STS inhibitor, exhibited an almost identical affinity for STS and also bound in a non-competitive manner. It is suggested that these compounds prefer to bind in a hydrophobic tunnel close to the entrance to the active site. The coumarin and chromenone boronic acids were modest inhibitors of STS with IC(50)s of 86 and 171microM, respectively. Surprisingly, replacing the boronic acid group of the chromenone derivative with an OH group yielded a good reversible, mixed type inhibitor with a K(i) of 4.6microM. Overall, these results suggest that the boronic acid moiety must be attached to a platform very closely resembling a natural substrate in order for it to impart a beneficial effect on binding affinity compared to its phenolic analog.

Boronic Acids↗

RNA editing by ADAR2 is metabolically regulated in pancreatic islets and beta-cells.

RNA editing via the conversion of adenosine (A) to inosine (I) is catalyzed by two major families of adenosine deaminases acting on RNA (ADARs), ADAR1 and ADAR2. This genetic recoding process is known to play essential roles in the brain, due in part to changes in functional activities of edited neurotransmitter receptors and ion channels. Little is known, however, about the physiological regulation and function of A to I RNA editing in peripheral tissues and other biological processes. Here, we report that both ADAR1 and ADAR2 are expressed in the murine pancreatic islets, and ADAR2 is primarily localized in the islet endocrine cells. In contrast to ADAR1, ADAR2 transcripts in the pancreatic islets exhibit a nearly 2-fold increase in insulin-resistant mice chronically fed a high fat diet. Concurrent with this diet-induced metabolic stress, RNA editing in the islets is dramatically enhanced for the RNA transcripts encoding the ionotropic glutamate receptor subunit B. Moreover, ADAR2 protein expression is repressed in the islets under fuel deficiency condition during fasting, and this repression can be completely reversed by refeeding. We also show that, specifically in pancreatic beta-cell lines, not only the expression of ADAR2 but also the glutamate receptor subunit B editing and ADAR2 self-editing are markedly augmented in response to glucose at the physiological concentration for insulin secretion stimulation. Thus, RNA editing by ADAR2 in pancreatic islets and beta-cells is metabolically regulated by nutritional and energy status, suggesting that A to I RNA editing is most likely involved in the modulation of pancreatic islet and beta-cell function.

Adenosine Deaminase↗

[Expressed genes related to adolescent idiopathic scoliosis in spinal facet].

OBJECTIVE: To construct a subtractive cDNA library from the spinal facets of adolescent idiopathic scoliosis (AIS) patient with suppression subtractive hybridization (SSH) technique and clone differentially expressed genes related to AIS in spinal facet. METHODS: mRNA was isolated separately from the convex side and concave side apical facets of an adolescent idiopathic scoliosis patient. Moreover, single-strand (ss) and double-strand (ds) cDNAs were synthesized in turn using SMART PCR cDNA synthesis technology. (ds) cDNAs then were digested with Rsa I and divided into two groups, and ligated to the specific adaptor 1 and adaptor 2R, respectively. After cDNAs were hybridized with each other twice and underwent two rounds of nested PCR, the PCR products were ligated with pDrive Cloning Vectors to set up the subtractive library. Sequence analysis was performed and the acquired data were aligned against the GeneBank nucleotide database. Furthermore, the spinal facets of 11 IS patients were collected. The techniques of immunohistochemistry and in situ hybridization were adopted in order to approve the gene expression difference. The images of immunohistochemistry and in situ hybridization were input to the image analysis system and were analyzed semi-quantitatively. RESULTS: A cDNA subtractive library of AIS in spinal facet was set up successfully with high subtractive efficiency. Beta2 Microglobulin (beta 2M) and calgranulin A (S100A8) were identified as highly differentially expressed genes in this study. CONCLUSION: All results confirm the effectiveness and sensitivity of SSH in detecting differentially expressed genes from a small amount of clinical samples. Information about such alterations in gene expression can be useful for elucidating the genetic events in the development of AIS.

Adolescent↗

Pediatric intractable epilepsy syndromes: reason for early surgical intervention.

Drug-resistance in several childhood epilepsy syndromes is common, and these patients may tolerate epilepsy surgery. In this study, the surgical outcomes of 24 pediatric patients with various intractable epilepsy syndromes and three patients with tuberous sclerosis were examined at Xinqiao hospital between 1997 and 2004. The study included nine cases of Lennox-Gastaut syndrome, two cases of Rasmussen's syndrome, one case of Sturge-Weber syndrome, three cases of West syndrome, three cases of tuberous sclerosis and nine cases of mesial temporal lobe epilepsy syndrome. In each case, different surgical procedures were performed according to preoperative evaluation and ECoG. At an average of 4.5 years after surgery, 14 out of 27 patients (51.9%) had an Engel Class I outcome after surgery, and an additional eight patients (29.6%) had rare seizure (Engel ClassII). Three patients showed a significant decrease in seizure frequency (Engel Class III). The mean IQ increased from 61.4+/-12.2 to 75.0+/-11.0, and greater IQ increase was seen in patients with shorter seizure history and drug-resistance. Temporary complications were observed in four patients and there were no deaths. In conclusion, early surgical intervention in pediatric intractable epilepsy syndromes may results in a favorable outcome in a high percentage of cases and may provides an important opportunity to prevent irreversible decline in intelligence and disability.

Adolescent↗

An optimal pooling strategy applied to high-throughput screening for rare marker-free transformants.

An optimal pooling system, called Accurate and Fast Target Screening, has been developed for high-throughput identifying the rare marker-free transformants. This system can identify targets between 10- and 100-fold more efficiently than analysis of individual samples. By calculating the efficiency for different proportions of targets and the optimal group size in a worst case scenario, we are able to estimate an upper limit for the number of tests that are required. The application of this system to determine the transgene in an artificially constructed population of transgenic and non-transgenic wheat lines successfully identified the 10 positive samples located randomly with 990 negative samples using only 92 PCR reactions. The same approach was also applied to determine transgene expression by SDS-PAGE of seed proteins. This system gives unambiguous positive or negative results and should facilitate marker-free transformation.

Algorithms↗

[Expression of calmodulin in the articular process of vertebrae of adolescent idiopathic scoliosis patients].

OBJECTIVE: To investigate the expression of calmodulin in the articular process of vertebrae of adolescent idiopathic scoliosis (AIS) patients and congenital scoliosis (CS) patients and its possible effects on the pathogenesis of AIS. METHODS: Nine AIS patients aged 13 - 17, and 10 patients of CS, aged 11 - 18, underwent orthopedic surgery. Specimens of the inferior articular process from the apical vertebrae and upper end vertebrae discarded during operation were collected from the AIS patients and specimens of the. Inferior articular process from the upper end vertebrae wee collected from the CS patients. Immunohistochemistry was used to detect the protein expression of calmodulin, and in situ hybridization was adopted to examine the mRNA expression of calmodulin. RESULTS: Calmodulin were expressed in the bone cells. There was no difference between the expression of calmodulin at the concave side and convex side, or between the apical vertebrae and end vertebrae. The expression of calmodulin of the AIS group was significant lower than that of the CS group (P < 0.05). The result of immunohistochemistry and that of in situ hybridization corresponded with each other. CONCLUSION: The expression of calmodulin in the articular process of vertebrae of AIS patients is low, suggesting that it plays a role in the development and progression of AIS.

Adolescent↗