Plasma expression levels of MEG3 and UCA1 in patients with acute myocardial infarction and their clinical significance / 心血管康复医学杂志

Objective :To explore plasma expression levels of human maternal expression gene 3 (MEG3) and urotheli-al carcinoma antigen 1 (UCA1) in patients with acute myocardial infarction (AMI) and their clinical significance . Methods : A total of 90 AMI patients treated in our hospital were enrolled as AMI group ,and 50 healthy subjects un-dergoing physical examination in our hospital simultaneously were treated as healthy control group .Plasma expres-sions of MEG3 and UCA1 ,serum levels of creatine kinase isoenzyme MB (CK-MB) and cardiac troponin I (cTnI) were observed and compared between healthy control group and AMI group on 1h ,3h ,6h and 12h after onset . Spearman correlation analysis was used to analyze their correlation .Results : Compared with healthy control group , there was significant rise in plasma MEG3 expression [ (0-002 ± 0-001) vs .0-017 ± 0-003)] ,and significant reduc-tion in UCA1 expression [ (0-027 ± 0-005) vs .(0-017 ± 0-002)] in AMI group on 1h after onset , P＝0-001 both ;after 3h ,there were significant rise in serum levels of CK-MB [ (20-01 ± 3-05) IU／L vs.( (32-10 ± 4-40) IU／L] and cTnI [ (1-01 ± 0-87) ng／L vs.(2-10 ± 0-91) ng／L] in AMI group , P＝0-001 all.Spearman correlation analy-sis indicated that in plasma MEG3 expression was significant positively correlated with serum CK-MB and cTnI levels ( r＝0-351 ,0-368 , P＜0-05 both) ,and plasma UCA1 expression was significant inversely correlated with serum levels of CK-MB and cTnI (r＝ －0-416 ,－0-425 , P＜0-01 both) AMI patients .Conclusion : Plasma MEG3 level significant rises ,and UCA1 level significantly reduces in AMI patients during early onset period .Both are signifi-cantly correlated with CK-MB and cTnI levels ,which may be used as new markers diagnosing early AMI .

Inhibitory Effect of NPM Gene Knockdown on Proliferation of Chronic Myeloid Leukemia Cell Line K562 and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the role of nucleophosmin (NPM) in the proliferation of chronic myeloid leukemia cells (K562 cells) and its mechanism by RNAi technology.@*METHODS@#shRNA was used to inhibit the expression of NPM. The expression of NPM gene was detected by real-time quantitative PCR. The effect of inhibiting NPM gene on cell proliferation was detected by MTS assay. Change of cell cycle was detected by flow cytometry. Western blot was used to detect the expression of cell cycle-related proteins.@*RESULTS@#The shRNA lentiviral vector targeting at NPM gene was successfully constructed and used to transfect the K562 cells. The results showed that compared with the control groups, suppression of NPM gene expression in K562 cells could inhibit the cell proliferation and decrease the cell colony formation. Moreover, interference of NPM gene could prolong G/G phase and arrest cell cycle, which may be related to the down-regulation of NPM gene expression and activation of p21 protein expression, thereby inhibited the formation of CDK2/ Cyclin E complex.@*CONCLUSION@#Down-regulation of NPM gene expression in K562 cells can induce cell cycle arrest and inhibit cell proliferation.

Therapeutic effect of levosimendan combined milrinone on severe refractory heart failure and its influ-ence on serum levels of BNP and UA / 心血管康复医学杂志

Objective:To study therapeutic effect of levosimendan combined milrinone on severe refractory heart failure (SRHF) and its influence on serum levels of brain natriuretic peptide (BNP) and uric acid (UA).Methods:A total of 156 SRHF patients were enrolled,randomly and equally divided into levosimendan group and milrinone group,both groups re-ceived corresponding medication based on routine treatment for 7d.Heart rate,blood pressure,serum BNP and UA levels,left ventricular end-systolic dimension (LVESd),left ventricular end-diastolic dimension (LVEDd) and left ventricular e-jection fraction (LVEF) before and after treatment,total effective rate and incidence of adverse reactions were observed and compared between two groups.Results:Compared with milrinone group after treatment,there were significant reduc-tions in heart rate [ (73.79 ± 7.61) beats/min vs.(70.39 ± 7.45) beats/min],systolic blood pressure [ (128.84 ± 13.11) mmHg vs.(121.86 ± 12.53) mmHg],scores of lung wet rales [ (2.05 ± 0.33) scores vs.(1.53 ± 0.21) scores],difficulty breathing [ (2.11 ± 0.36) scores vs.(1.60 ± 0.25) scores] and lower extremity edema [ (2.03 ± 0.34) scores vs.(1.50 ± 0.18) scores],serum levels of BNP [ (459.62 ± 46.27) μg/L vs.(248.73 ± 25.91) μg/L] and UA [ (355.97 ± 36.47) μmol/L vs.(282.75 ± 28.61) μmol/L],LVESd [ (41.62 ± 4.52) mm vs.(36.87 ± 3.71) mm] and LVEDd [ (51.89 ± 5.37) mm vs.(47.85 ± 4.83) mm],and significant rise in 24h urine volume [ (3204.59 ± 321.52) ml vs.(3695.78 ± 370.62) ml] and LVEF [ (42.36 ± 4.31)% vs.(47.85 ± 4.86)%] in levosimendan group,P＜0.01 all.Total effective rate of levosimendan group was significantly higher than that of milrinone group (89.74% vs.71.79%),and incidence rate of adverse reactions was significantly lower than that of milrinone group (5.13% vs.28.21%),P＜0.01 both.Conclusion:Levosimendan therapy can significantly reduce serum BNP and UA levels,improve cardiac function in SRHF patients.It possesses significant therapeutic effect,and it＇s safe and reliable,which is better than milrinone.

Effects of emodin on apoptosis and cell cycle related genes in U937 cells / 中国实验血液学杂志

This study was aimed to observe the effects of emodin on apoptosis and cell cycle related genes in human myeloid leukemia cell line U937 cells. U937 cells were exposed to 60 µmol/L emodin for 24, 48, 72 h. The expressions of C-MYC, h-TERT, PIM-2, Survivin, wild type P53, P21, TGF β-1 and MCL-1 genes before and after treatment with emodin were determined and quantitated by using reverse transcriptase-polymerase chain reaction (RT-PCR). The results showed that the expressions of C-MYC, h-TERT, PIM-2, Survivin in treated U937 cells decreased, but the expressions of WTp53, P21 and TGFβ1 increased, while the expression of MCL-1 gene had no obvious change. It is concluded that multiple pathways may be involved in the processes of emodin-induced U937 cell apoptosis.

Reversing effects of emodin on multidrug resistance in resistant HL-60/ADR cells / 中国实验血液学杂志

This study was aimed to investigate the reversing effects of emodin on multidrug resistance (MDR) in resistant HL-60/ADR cells, and to explore the underlying mechanisms. The MTT assay was used to assess the chemoresistance of HL-60/ADR cells to emodin and 8 chemotherapeutic agents commonly used in clinic. The reversal effects of emodin on MDR of HL-60/ADR cells were also evaluated by MTT method. DNA ploidy analysis and DNA Ladder assay were used to detect apoptosis-induced effects on HL-60/ADR cells via the adriamycin (ADR) and emodin combination. The expression changes of the drug resistance-associated genes and proteins were detected by RT-PCR and Western Blot respectively. The intracellular accumulation and subcellular distribution of ADR and DNR were measured by flow cytometry and confocal laser scanning microscopy. The results showed that emodin inhibited HL-60/ADR cell proliferation with an average IC50 value of 24.09 ± 1.72 µmol/L, which was similar to that of the parental HL-60 cells (average IC50 = 23.18 ± 0.87 µmol/L). HL-60/ADR cells were resistant to a variety of chemotherapeutic agents, such as ADR, DNR, VP16, VCR,Ara-C, HHT, MTZ and THP. The reversal multiple were between 1.58 and 4.12 after the treatment with low concentration of emodin combined with the above mentioned different agents. The combination of ADR with emodin showed the best reversal effects, and the typical hypodiploid peak (apoptotic peak) and DNA ladder could be detected after the co-treatment.In addition, emodin down-regulated the mRNA and protein expression levels of MRP1, TOPOIIβ, GST π and BCL-2. Furthermore, the addition of emodin enhanced ADR and DNR intracellular accumulation and subcellular distribution in HL-60/ADR cells in dose-dependent manner. It is concluded that the emodin shows reversing effects on the multidrug resistant HL-60/ADR cells, possibly via decreasing the expression levels of drug resistance-associated genes, increasing the intracellular accumulation of chemotherapeutic agents and activating the apoptosis pathway.

Synthesis of emodin derivatives and their inhibiting effects on proliferation of leukemia cell lines / 中国实验血液学杂志

The aim of this study was to explore the inhibitory effect of newly synthesised emodin derivatives on the proliferation of leukemia cell lines and to select the most effective one from these emodin derivatives for further research. Emodin derivatives were synthesized by modifying the structure of emodin. MTT method was used to detect the proliferative inhibition in leukemia cell lines treated with emodin derivatives. The results showed that the half inhibitory concentration (IC50) for K562 cells treated with emodin derivatives E10-19 for 48 h was 0.84 - 12.01 µmol/L. E19 displayed the best anti-proliferative activity, while E16 and E17 did not show effects on K562 cells. Emodin derivative E19 was chosen for treating U937, NB4, Molt-4 and CA-46 cells, their IC50 for 48 h were 0.85, 0.9, 0.76, 0.8 µmol/L respectively. The IC50 of E19 for LQ2 cells was 3.60 µmol/L, and the IC50 range of E19 for normal human peripheral blood mononuclear cells at 48 h was 4.01 - 4.78 µmol/L. It is concluded that emodin derivative E19 can strongly inhibit the growth of leukemia cells and its inhibiting effect on proliferation of leukemia cells has a certain specificity. The specific mechanism of E19 anti-leukemia effect should be further studied.

A case of Richter syndrome transformed from chronic lymphocytic leukemia with karyotype aberration of trisomy 12 / 中国实验血液学杂志

This study was aimed to investigate the relationship between Richter's syndrome (RS) transformation and clinical characteristics as well as karyotype of patient with chronic lymphocytic leukemia (CLL). By the follow-up of a patient with CLL, the clinical characteristics, karyotype, treatment pattern and its effect, as well as disease progression were monitored regularly with serological test, flow cytometry and FISH technique. The results indicated that the patient typically presented with history of CLL at initial diagnosis, with expression of CD5(+), CD19(+) and CD23(+), Binet stage C, as well as karyotype aberration of trisomy 12, and poorly responded to 4 cycles of standard chemotherapy of FCR regimen. The disease progression was confirmed at 5 months with the symptoms of fever in the absence of infection, elevated lactate dehydrogenase level and rapidly enlarging lymphnodes which showed typically diffuse large B cell lymphoma by the biopsy. It is concluded that karyotype aberration of trisomy 12 is one of the risk factors for RS transformation, and treatment pattern of the patient with CLL may be associated with the transformation of RS.

Clinical analysis on 40 patients with chronic lymphocytic leukemia / 中国实验血液学杂志

This study was aimed to analyze the clinical and laboratorial characteristics of patients with chronic lymphocytic leukemia (CLL), as well as their relationship with outcomes of patients. The clinical and laboratorial data of 40 CLL patients admitted from 2004 to 2010 in our hospital were analyzed retrospectively. The results indicated that the most of CLL attacked the elderly male patients with median age 66 (from 42 to 80). Flow cytometric analysis showed that 25 cases were positive for typical immunophenotype of CLL. On the other hand, all the patients clearly expressed CD19 and CD5, 7 cases (17.5%) and 14 cases (35%) were positive for the expression of CD38 and Zap70 respectively. 8 cases harbored a mutated immunoglobulin heavy-chain (VH) gene, among them 4 cases belong to VH3 family. Interphase FISH analysis showed that P53 deletion, RB1 deletion, trisomy 12 and normal chromosome were detected in 6, 3, 1, and 5 cases, respectively. The median PFS in 31 patients received treatment of fludarabine based chemotherapy was 48 months (95%CI: 39 - 57 months), among them 27 cases (87.1%) achieved CR + PR. While PFS was 14 months (95%CI: 10 - 18 months, P < 0.001) in 9 patients received other treatment regimen, out of them only 3 cases (33.3%) achieved CR + PR. Patients with normal level of serum β2-microglobulin at diagnosis showed significantly higher overall survival (78%, 95%CI: 69% - 87%) in 36 months than those with abnormal level of serum β2-microglobulin (47%, 95%CI: 35% - 59%, P = 0.004). Significant difference in the rate of CR + PR was noted in the Zap70 positive group (50%) and in negative group (88.5%, P = 0.006). All of 8 patients with IgVH mutation displayed CR after treatment, while 4 cases (66.7%) archived CR among 6 patients without IgVH mutation. It is concluded that CLL is characterized by high heterogeneity in both clinical features and molecular markers, which are associated with prediction of outcomes for patients. The treatment with fludarabine-based chemotherapy results in a major benefit and long survival for patients with CLL.

Clinical observation on children-sized fibreoptic bronchoscope usage in whole-lung lavage / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the effect of children-sized fibreoptic bronchoscope in improving the safety of whole-lung lavage (WLL).</p><p><b>METHOD</b>Patients from May 2006 to May 2010 using children-sized fibreoptic bronchoscope to assistant the location were assigned to fibreoptic bronchoscope group. Patients from May 1998 to Nov 2004 using traditional stethoscope to help intubation were assigned to control group. The adverse reactions and complications were compared.</p><p><b>RESULT</b>There were liquid leakage 1 case (0.96%), hypoxia 3 cases (2.88%) and liquid retained over 1000 ml 15 cases (14.42%) in fibreoptic bronchoscope group. In contrast, liquid leakage 24 cases (6.38%), hypoxia 42 cases (11.17%) and liquid retained over 1000 ml 135 cases (35.90%) happened in control group. The differences between the two groups were significant (P<0.05, P<0.01).</p><p><b>CONCLUSION</b>Using children-sized fibreoptic bronchoscope in WLL can promote the situation of double-lumen tube, help separation the two lungs, decrease complications and improving safety.</p>

Construction of eukaryotic vector of small hairpin interfering RNA against NYD-SP5 / 中华男科学杂志

<p><b>OBJECTIVE</b>NYD-SP5 is a newly cloned gene highly expressed in human testes, which consists of 3 598 nucleotides including a 1 027-amino acid open reading frame. It is a human-mouse homologous gene. The domain analysis indicated that the NYD-SP5 protein is a transmembrane protein. This study aimed to design and establish recombinant plasmids of small hairpin interfering RNA (shRNA) against NYD-SP5, and to pave the way for the analysis of the function of NYD-SP5 in the testis using the transgenic mouse model.</p><p><b>METHODS</b>Four sequences of oligonucleotides with the small hairpin structure were designed based on the NYD-SP5 mRNA sequence. Recombinant plasmids were constructed by cloning these oligonucleotides into pGPU6/GFP/Neo vectors. Interfering plasmids against GAPDH were established as positive controls and those targeting non-specific genes used as negative controls. The positive constructs were verified by enzyme digestion and sequencing.</p><p><b>RESULTS</b>Plasmid screening and sequencing showed the sequences of the recombinant plasmids to be the same as the shRNA transcribed sequences, which indicated the successful establishment of the recombinant vectors.</p><p><b>CONCLUSION</b>The shRNA expression vector targeting NYD-SP5 could be established successfully.</p>

Effect of emodin on proliferation inhibition and apoptosis induction in leukemic K562 cells / 中国实验血液学杂志

The study was aimed to investigate the effects of emodin on proliferation inhibition and apoptosis induction in human chronic myeloid leukemia cell line K562 cells, and to explore the role of P210 protein and activation of caspase 3 in these processes. K562 cells were exposed to emodin at different doses. The proliferation inhibition was detected by MTT assay and colony formation test. The ability of emodin to induce apoptosis and DNA fragmentation were examined by flow cytometry. The expressions of P210, procaspase-3 and PARP protein were determined by Western blot. The results indicated that the emodin remarkably inhibited the K562 cell proliferation, with IC(50) value of 38.25 micromol/L after treatment for 48 hours. Meanwhile induced apoptosis, Annexin V-FITC positive cells, sub-G(1) apoptotic peak and DNA fragmentation in K562 cells confirmed that emodin induced apoptosis in K562 cells in dose-dependent manner. Western blot results showed that emodin inhibited phosphorylation of P210 protein in K562 cells and down-regulated the expression levels of P210. The procaspase-3 level in treated K562 cells decreased with increased expressions of PARP in time-dependent manner. It is concluded that the emodin efficiently inhibits growth and induces apoptosis of K562 cells, while the inhibition of phosphorylation of P210 protein, down-regulation of P210 protein expression and activation of caspase-3 may be involved in these processes.

Decreased expression of DICER1 in gastric cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The role of epigenetics in gene expression regulation and development significantly enhances our understanding of carcinogenesis. All the tumor related genes may be the target of epigenetical or genetic regulation. We selected some epigenetically regulated genes for cDNA array analysis and observed variability in the expression of the DICER1 gene in distinct stages of gastric cancer. The aim of this study was to assess the correlation between the expression of DICER1, an epigenetically regulated gene, and gastric cancer.</p><p><b>METHODS</b>To detect the expression of 506 tumor-associated genes, including DICER1, in the matched cancerous mucosa, pre-malignant lesion (adjacent mucosa), non-cancerous gastric mucosa and distant lymphocyte metastatic lesion in 3 cases of gastric cancers using cDNA array. DICER1 mRNA expression and DICER1 protein expression were further analyzed by Real-time PCR and Western blot in 32 cases of progressive gastric cancer. DICER1 protein expression was also detected in 33 early and 30 progressive gastric cancers by the immunohistochemistry (IHC) method.</p><p><b>RESULTS</b>In 3 cases of gastric cancer cDNA array showed dramatically decreased expression of DICER1 in pre-malignant lesion, cancerous mucosa and distant lymphocyte metastatic lesions compared with matched noncancerous gastric mucosa, pre-malignant lesion and cancerous mucosa. Real-time PCR results showed that the expression level of DICER1 mRNA in gastric cancer was significantly down-regulated compared to normal gastric tissue (P < 0.05). The IHC assay also showed that the expression of DICER1 was significantly decreased in progressive gastric cancer. Among the 63 cases of gastric cancers, 13/33 early (39.4%) and 19/30 (63.3%) progressive cancers showed negative expression of DICER1 (50.8%). The difference in expression of DICER1 between early and progressive gastric cancers was significant (P < 0.01). The result of Western blotting showed that DICER1 protein was down-regulated significantly in advanced gastric cancer (P < 0.05).</p><p><b>CONCLUSIONS</b>DICER1 expression is decreased during the progression of gastric cancer, especially in progressive gastric cancers, which indicating DICER1 may play an important role in the development of cancer and the epigenetical regulation involved.</p>

Inhibitory effects of emodin on drug-resistant HL-60/ADR cell proliferation and its induction of apoptosis / 中国实验血液学杂志

The study was aimed to investigate the effects of emodin on the proliferation and apoptosis of adriamycin-resistant HL-60/ADR cells, and to explore the underlying mechanism. The cell viability and colony formation were detected by MTT assay and colony formation assay respectively. Apoptotic cells were tested by means of cell cycle analysis, mitochondrial transmembrane potential levels, caspase-3 activity detection, Annexin V FITC/PI staining and TUNEL labeling. RT-PCR was used to analyze the bcl-2 and c-myc mRNA expressions. The protein expressions of Bcl-2, c-Myc and caspase-3 precursor were determined by Western blot. The results showed that HL-60/ADR cell growth was significantly inhibited by emodin in dose and time dependent manners. Cell colony formation obviously decreased with IC50 5.79 micromol/L. G0/G1 phase cell population increased while G2/M phase cells decreased in 40 and 80 micromol/L groups compared with control group (p < 0.01), and no significant difference of cell cycle was observed in 20 micromol/L group (p > 0.05). The typical hypo-diploid peak (apoptotic peak) appeared in each dose group. The levels of mitochondrial transmembrane potential of HL-60/ADR cells decreased and caspase-3 activity increased when incubated with emodin for 12 and 24 hours respectively. Apoptosis occurred in a dose-dependent manner, and its earlier and later stages were identified by Annexin-V FITC/PI staining and TUNEL labeling methods respectively. The expressions of bcl-2, c-myc mRNA and Bcl-2, c-Myc, caspase-3 precursor protein were all down-regulated in a time-dependent manner after treatment with emodin at different times. It is concluded that emodin efficiently inhibits growth and induces apoptosis on HL-60/ADR cells, which may be related with the down-regulation of mitochondrial transmembrane potential and expressions of bcl-2 and c-myc, as well as up-regulation of caspase-3 activity.

Emodin induces leukemic HL-60 cells apoptosis probably by inhibiting Akt signal pathway / 药学学报

This study is to investigate the effect of emodin on inducing human myeloid leukemia cell line HL-60 apoptosis and the role of Akt signal pathway in the apoptosis. HL-60 cells were exposed to various dosages of emodin. MTT assay was used to detect HL-60 cell proliferation. Distribution of HL-60 cells in cell cycle was analyzed by flow cytometry and cell apoptosis was observed by MitoCapture apoptosis detection. The protein expressions of Akt signal pathway were detected by Western blotting. The result showed that emodin remarkably inhibited the cell proliferation. The IC50 value for 48 h treatment was about 20 micromol x L(-1). Apoptosis in HL-60 cells could be efficiently induced by emodin in a dose dependent manner and cells were arrested at G0/G1. The expressions of Akt, p-Akt, IkappaB-alpha, p-IkappaB-alpha, p65, p-p65, mTOR and p-mTOR in Akt signal pathway were downregulated after emodin treatment. It can be concluded that emodin could efficiently induce growth inhibition and apoptosis in HL-60 cells. Akt signal pathway may be involved in this process.

Gene expression patterns in gastric cancer / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify gene expression patterns in distinct stages of intestinal-type gastric cancer(GC).</p><p><b>METHODS</b>Gene expression patterns of distinct stages of intestinal-type GC samples from 3 patients were compared with cDNA microarray, which contained 576 genes. There were 506 target genes, which included 51 genes identified from our previous experiment with suppression subtractive hybridization(SSH) and other 455 genes chosen for their important roles in cancers. Hierarchical clustering was performed to clarify genes in association with distinct stages of GC.</p><p><b>RESULTS</b>One hundred and eighty-one differentially expressed genes with average Cy5:Cy3 ratios higher than 2.0 or lower than 0.5 in at least one stage of GC were identified by cDNA microarray. Among them, 48 genes were up-regulated and 133 down-regulated. Hierarchical clustering analysis separated the differentially expressed genes in different stages of GC into 5 main characteristic groups. Some important differentially expressed genes in different stages of GC were identified, such as SEC23IP, LIPF, ES(BQ291520), SLC5A1, PG(encoding similar to pepsin A precursor), CXCR4, DICER1, SH3GL2, and IGF2R.</p><p><b>CONCLUSION</b>The differentially expressed gene patterns and some important genes were identified, which might be useful in further study on carcinogenesis, progression and metastasis of intestinal-type GC.</p>

Rejuvenation of the forehead/temple by limited incision / 中华整形外科杂志

<p><b>OBJECTIVE</b>To utilize multiplane, subcutaneous and subperiosteal, dissection through small incisions in scalp to rejuvenate aging signs of forehead/temple.</p><p><b>METHODS</b>Forehead: We make four small incisions in scalp, widely separate tissues between subperiosteum skin and frontal muscle to form galea frontal muscle-periosteum flap, the flap is tightened and sutured with the galea at the posterior border of the incision. This method avoid to excise scalp. Temple: there are two small incisions in the scalp of temple at each side. Superficial temple fascia and orbicularis oculi muscle as well is dissected from deep temporal fascia and skin respectively, then tightened and sutured with the superficial part of deep temporal fascia, excising scalp is not necessary.</p><p><b>RESULTS</b>Twenty patients have received this treatment, the effects are satisfying.</p><p><b>CONCLUSIONS</b>Multiplane dissection through small incisions can remove wrinkles of skin, correct the prolapse of eyebrow and avoid the complications of coronary incision. This approach brings more rapid recovery of patients, it is safe and affective.</p>

Screening and analysis of associated genes in the carcinogenesis and progression of gastric cancer / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To screen and analyze the important associated genes in different stages of gastric cancer.</p><p><b>METHODS</b>Using suppression subtractive hybridization (SSH) to screen differentially expressed genes; detecting the expression of genes in different stages of gastric cancer with dot blot hybridization; and verifying the results with semi-quantitative reverse transcriptase-polymerase chain reaction(RT-PCR).</p><p><b>RESULTS</b>Twenty-six differentially expressed gene fragments were obtained by means of SSH. Among them,24 were known genes, 1 was a new expressed sequence tags(EST), and 1 was a hypothetical gene. The results of dot blot hybridization demonstrated that the expressions of Annexin A2, RPS29, RPS12 etc. in dysplasia were higher than those in normal mucosa; the expressions of RPS12 etc.in early cancer were higher than those in normal mucosa;the expressions of cytochromosome C oxidase II, ferritin light chain, RPS12 etc. in advanced gastric cancer and lymph node metastases were consistently higher than those in normal mucosa. The expression of proteasome 26S subunit gene in advanced gastric cancer was higher than that in normal mucosa. The expression of RPS12 was consistently higher in different stages of gastric cancer. It was demonstrated by RT-PCR that the expression of RPS12 in gastric cancer was higher than that in normal mucosa.</p><p><b>CONCLUSION</b>The authors have identified some important genes that might be involved in the carcinogenesis and progression of gastric cancer, and RPS12 may play more important roles in gastric cancer.</p>

Cloning associated genes using microdissection-cDNA PCR-SSH in gastric dysplasia / 中国医学科学院学报

<p><b>OBJECTIVE</b>To construct cDNA subtracted libraries from gastric dysplasia and further screen differentially expressed genes.</p><p><b>METHODS</b>Relatively pure dysplasia and normal tissue were procured by manual microdissection, and amplified by cDNA-PCR, which was used to carry on for suppression subtractive hybridization (SSH). Subtracted cDNA fragments were linked with vector, cloned, screened, sequenced, and made homologous search. Differentially expressed fragments were verified by dot hybridization.</p><p><b>RESULTS</b>Two subtracted cDNA libraries were constructed. Among 26 sequenced clones, 15 fragments corresponded to known genes, 3 fragments were known EST and 8 fragments were unknown EST (GenBank BQ164614-BQ164616, BQ291516-BQ291520). Fifteen fragments were verified to be differentially expressed in gastric dysplasia.</p><p><b>CONCLUSIONS</b>Subtracted cDNA libraries from gastric dysplasia are constructed using combination of microdissection-cDNA PCR and SSH setup in our laboratory. Some fragments have been screened and verified to help to search for novel associated genes with gastric carcinogenesis.</p>

Differentiation of neural stem cells transplanted into ventral horn of spinal cord after brachial plexus avulsion / 中华显微外科杂志

Objective To study the survival,migration and differentiation of the neural stem cells which transplanted into ventral horn of spinal cord after brachial plexus avulsion.Methods Neural stem ceils isolated from spinal cord of neogenetic rats and cultured,expanded,labeled by BrdU before transplanted. Twenty adult healthy SD rats preformed as the model of brochial plexus avulsion(Roots C_(5～7)),then transplan- rod stem ceils into the C_6 ventral horn of spinal cord.On 1,2,4,8,12 weeks postoperatively,immunohisto- chemistry assay were carried out in the spinal cord.Results Transplanted into ventral horn of spinal cord after brachial plexus avulsion.Neural stem cells can survive,migrate for at least one segment of spinal cord and differentiate to neurons and astrocytes.The differentiation of stem cells were time-depends.Conclusion Neural stem cells can survive,migrate and differentiate after transplanted into ventral horn of spinal cord in the rats which suffered from brachial plexus avulsion.