Effects of environmental hypothermia on hemodynamics and oxygen dynamics in a conscious swine model of hemorrhagic shock / 世界急诊医学杂志（英文）

BACKGROUND: Hypothermia is associated with poor outcome in trauma patients; however, hemorrhagic shock (HS) model with anesthetized swine was different from that of clinical reality. To identify the effects of environmental hypothermia on HS, we investigated hemodynamics and oxygen dynamics in an unanesthetized swine model of HS under simulating hypothermia environment.METHODS: Totally 16 Bama pigs were randomly divided into ambient temperature group (group A) and low temperature group (group B), 8 pigs in each group. Venous blood (30 mL/kg) was continuously withdrawn for more than 15 minutes in conscious swine to establish a hemorrhagic shock model. Pulmonary arterial temperature (Tp), heart rate (HR), mean arterial pressure (MAP), pulmonary arterial pressure (PAP), pulmonary arterial wedge pressure (PAWP), central venous pressure (CVP), cardiac output (CO), hemoglobin (Hb), saturation of mixed venous blood (SvO2) and blood gas analysis were recorded at the baseline and different hemorrhagic shock time (HST). The whole body oxygen delivery indices, DO2I and VO2I, and the O2 extraction ratio (O2ER) were calculated.RESULTS: Core body temperature in group A decreased slightly after the hemorrhagic shock model was established, and environmental hypothermia decreased in core body temperature. The mortality rate was significantly higher in group B (50％) than in group A (0％). DO2I and VO2I decreased significantly after hemorrhage. No difference was found in hemodynamics, DO2I and VO2I between group A and group B, but the difference in pH, lactic acid and O2ER was significant between the two groups.CONCLUSION: Environmental hypothermia aggravated the disorder of oxygen metabolism after hemorrhagic shock, which was associated with poor prognosis.

Short-term outcomes of radical resection of rectal carcinoma: hand-assisted laparoscopy versus traditional laparoscopic approach / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To study the feasibility of hand-assisted laparoscopic radical resection of rectal carcinoma and compare the short-term outcomes of HALS versus traditional laparoscopy approach.</p><p><b>METHODS</b>Clinical data of 42 cases of rectal carcinoma between January 2010 and March 2011 were enrolled in this study. Nineteen cases underwent HALS total mesorectal excision and 23 cases underwent traditional laparoscopy approach.</p><p><b>RESULTS</b>All the operations were successfully accomplished without conversions to open surgery. The mean operation time of the HALS group was shorter than that of the traditional laparoscopic group (152 min vs. 168 min, P=0.009). Incision length was significantly longer in the HALS group (5.6 cm vs. 4.5 cm, P=0.000). The median overall costs were lower in HALS group (26 000 RMB vs. 29 000 RMB, P=0.008). The number of lymph nodes in resected specimen, intra-operative blood loss, length of hospital stay, time to passage of flatus were comparable between the two groups.</p><p><b>CONCLUSIONS</b>Hand-assisted laparoscopic surgery has the advantages of laparoscopic surgery including minimal invasiveness, safety, and quicker postoperative recovery.</p>

Correlation of seven biological factors (Hsp90a, p53, MDM2, Bcl-2, Bax, Cytochrome C, and Cleaved caspase3) with clinical outcomes of ALK+ anaplastic large-cell lymphoma / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To explore correlation of seven apoptosis-related proteins (Hsp90a, p53, MDM2, Bcl-2, Bax, Cytochrome C, and Cleaved caspase3) with clinical outcomes of ALK+ anaplastic large-cell lymphoma (ALCL).</p><p><b>METHODS</b>Using immunohistochemistry and immunofluorescence double staining methods, the expressions of these seven apoptosis-associated proteins were studied to clarify their relationship with clinical outcomes of 36 ALK+ and 25 ALK-systemic ALCL patients enrolled between 1996 and 2006. The relationship of these apoptosis-regulating proteins with NPM-ALK status was also evaluated with the tyrosine inhibitor herbimycin A (HA) in vitro by immunocytochemistry, Western blotting and flow cytometric assays.</p><p><b>RESULTS</b>The presence of Hsp90α-, MDM2-, Bax-, Cytochrome C, and Cleaved caspase3-positive tumor cells was found significantly different in ALK+ and ALK-ALCLs, which was correlated with highly favorable clinical outcome. The Bcl-2- and p53-positive tumor cells were found in groups of patients with unfavorable prognosis. Inhibition of NPM-ALK by HA could reactivate the p53 protein and subsequent apoptosis-related proteins and therefore induced apoptosis in ALK+ ALCL cells.</p><p><b>CONCLUSION</b>Our results suggest that these seven proteins might be involved in apoptosis regulation and associated with clinical outcome of ALK+ systemic ALCLs. We also reveal a dynamic chain relation that NPM-ALK regulates p53 expression and subsequent apoptosis cascade in ALK+ ALCLs.</p>

Expression of ALK protein in large cell lymphoma with ALCL chromosome translocation in relation to prognosis / 中国实验血液学杂志

The aim of this study was to investigate the expression of anaplastic lymphoma kinase (ALK) protein resulted from chromosome translocation in anaplastic large cell lymphoma (ALCL) and its relationship with the age and prognosis of patients with ALCL. The tissue microarray including 30 cases of ALCL and 2 normal control tissues were established, the expression of anaplastic lymphoma kinase (ALK) protein was detected by immunohistochemistry, the statistical analysis of detected results was carried out by SPSS software. The results showed that the ALK protein was expressed negatively in 2 cases of primary skin ALCL, but in 20 out of 28 cases of systematic ALCL the ALK protein was expressed positively and mainly located in cytoplasm and/or nucleus (71.4%). Clinically, the patients with ALK expression were younger than those without ALK expression (p < 0.05). The prognosis of patients with ALK expression was better than those without ALK expression (p < 0.05). It is concluded that there is a high incidence of ALK expression in ALCL, especially in younger group. ALK expression may be an useful and independent marker for the differential diagnosis and prognosis evaluation of ALCL.

Detection of 3q27 chromosomal abnormality in diffuse large B cell lymphoma using FISH on cell microarray / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the association of 3q27 chromosome rearrangement with bcl-6 gene amplification and the molecular classification, therapeutic efficacies, and clinical stages in diffuse large B cell lymphoma (DLBC).</p><p><b>METHODS</b>The newly invented cell microarray was used to detect 3q27 chromosome rearrangement and bcl-6 gene amplification in 60 cases of DLBCL by fluorescence in situ hybridization (FISH). The molecular classification of germinal center B-cell-like (GCB) and non-germinal center B-cell-like (non-GCB) was investigated by analyzing the expression of CD20, CD10, bcl-6 and MUM1 simultaneously by immunohistochemical S-P method and tissue microarray. The information of therapeutic efficacies and clinical stages was obtained by analyzing clinical cases. The relationships among the factors were analyzed by statistics.</p><p><b>RESULTS</b>In 60 cases of DLBCL, 48.3%(29/60) were GCB and 51.7%(31/60) were non-GCB. The 3q27 chromosome rearrangement and bcl-6 gene amplification were present in 15 and 22 cases respectively. In 15 cases with 3q27 rearrangement, bcl-6 protein expression was positive in 3(20.0%), which was significantly different from that in cases without 3q27 rearrangement (P=0.017). In 60 cases of DLBCL, bcl-6 gene amplification was present in 22 cases, in which 5(22.7%) were GCB and 17(77.3%) were non-GCB, which was significantly different from that in cases without bcl-6 gene amplification (P=0.003). In 36 cases undergoing the normal CHOP program treatment, bcl-6 gene amplification was present in 15 cases and the rates of the complete remission, partial remission and no change were 4(26.7%), 4(26.7%) and 7(46.7%) respectively, and again it was significantly different from that in cases without bcl-6 gene amplification (P=0.016). There were no statistical significances among bcl-6 gene, BCL-6 protein expression, and clinical stages. Cases with BCL-6 protein positive and negative expression were not correlated with therapeutic efficacies and clinical stages.</p><p><b>CONCLUSION</b>There is lower expression of BCL-6 protein in cases with bcl-6 gene fragmentation. Cases with bcl-6 gene amplification are non-GCB with worse therapeutic results and later clinical stages. There may be other genes near chromosome 3q27 associated with DLBCL prognosis.</p>

Nuclear microarray combined with fluorescence in situ hybridization for detecting ALK gene translocation in paraffin-embedded anaplastic large cell lymphoma and its significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare the efficacy of nuclear microarray combined with fluorescence in situ hybridization (FISH) and immunohistochemistry in detecting ALK gene translocation and ALK fusion protein in anaplastic large cell lymphoma (ALCL).</p><p><b>METHODS</b>ALK gene translocation and ALK fusion protein in 17 paraffin-embedded ALCL specimens were detected using nuclear microarray combined with FISH and immunohistochemical straining, respectively.</p><p><b>RESULTS</b>The expression of ALK fusion protein was detected immunohistochemically with ALK antibody in 8 of the 17 specimens of systemic ALCL, including 4 with both nuclear and cytoplasmic positivity and 4 with only cytoplasmic positivity. Dual-color FISH identified 6 positive specimens, including the 4 specimens with both nuclear and cytoplasmic positivity as identified immunohistochemically, and 2 with immunohistochemical cytoplasmic positivity. FISH yielded negative results for the 2 specimens with immunohistochemical cytoplasmic positivity.</p><p><b>CONCLUSION</b>Nuclear microarray combined with FISH eliminated the cytoplasmic interference of the results of conventional FISH and provides a high-throughput platform for clinical detection with greater specificity than immunohistochemistry.</p>

Establishment of a whole-body visualization model of orthotopically implanted colorectal carcinoma and metastasis model in nude mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish an animal model visualizing orthotopic growth and metastasis of colorectal cancer.</p><p><b>METHODS</b>pEGFP-N1 plasmid was transfected into human colorectal carcinoma cell line SW480 so that the resultant SW480/EGFP cells emitted fluorescence that could be detected externally by fluorescence stereo microscope. SW480/EGFP cells were inoculated subcutaneously in nude mice, and the orthotopic tumor growth was evaluated in real time. Whole-body visualization models of orthotopically implanted colorectal carcinoma was established surgically, and the tumor growth and metastasis are evaluated by conventional pathological methods.</p><p><b>RESULTS</b>SW480/EGFP cells stably expressed high-levels of enhanced green fluorescent protein. Subcutaneous injection of SW480/EGFP cells resulted in tumor growth in nude mice, and the emitted fluorescence could be quantitatively detected and imaged with fluorescence stereo microscope to visualize real-time tumor growth. Visualization animal model was established successfully with surgical orthotopic implantation (SOI) of the tumor, and all mice survived. After two weeks, all the mice developed colorectal carcinoma without metastasis, but 4 weeks later, 75%percnt; of the mice developed peritoneal tumor metastasis and 50% had liver metastasis. The whole-body visualization animal model was successfully validated by pathological detection.</p><p><b>CONCLUSION</b>Whole-body visualization model of orthotopic and metastatic tumor growths provides a reliable means for observing the behavior of human colorectal carcinoma and can be helpful to study the growth and metastasis patterns of the cancer.</p>

Detection of t (14; 18) translocation and bcl-2 amplification in diffuse large B-cell lymphoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the role of t (14; 18) chromosomal translocation and bcl-2 amplification in classification, clinical staging and prognostic evaluation of diffuse large B cell lymphoma (DLBCL).</p><p><b>METHODS</b>Sixty cases of DLBCL were included in this investigation. Microdissection of the lymphoma tissue was performed. Tissue microarray and in-situ fluorescence hybridization technique were used to detect t (14; 18) and bcl-2 amplification. The phenotypes of either germinal center B-cell-like (GCB) or non-germinal center B-cell-like (non-GCB) were determined by immunohistochemistry including CD20, CD10, bcl-6 and MUM1 (S-P method) using the tissue microarray format. Clinical staging and therapeutic response were obtained by medical record review. The relationships among different parameters were analyzed by appropriate statistical methods.</p><p><b>RESULTS</b>Among 60 cases of DLBCL, bcl-2/IgH was positive in 10 cases and bcl-2 gene amplification was detected in 18 cases. Overall, 29 (48.3%) cases were GCB and 31 (51.7%) cases were non-GCB. The t (14; 18) was seen in 8 (80.0%) cases of GCB and 2 (20.0%) of non-GCB. The difference was statistical significance (P = 0.031). Over-expression of bcl-2 was seen in all cases having both t (14; 18) and bcl-2 gene amplification. Of thirty-six patients who underwent routine CHOP treatment, bcl-2 gene amplification was seen in 13 cases. In these cases, the rates of complete remission, partial remission and no change were 3 (23.1%), 4 (30.8%) and 6 (46.2%) respectively, and the clinical stages were stage I - II (1 case, 7.7%) and stage III - IV (12 cases, 92.3%). The clinical stages and therapeutic response were significantly different between the bcl-2 amplification cases and those without (P = 0.046 and P = 0.019, respectively).</p><p><b>CONCLUSIONS</b>T (14; 18) and/or bcl-2 gene amplification can lead to an over-expression of bcl-2 protein. The bcl-2 gene amplification correlates with worse therapeutic efficacies and advanced clinical stages. The reason for the correlation between bcl-2 over-expression and the prognosis is unclear, although it may be explained by different mechanisms that lead to bcl-2 over-expression. Detection of t (14; 18) chromosome translocation by FISH can be helpful in the classification of DLBCL.</p>

Expression and cloning of mCD99L2 gene from mouse B lymphoma cell line A20 and construction of its eukaryotic expression vector / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect and clone mCD99L2 gene from mouse B lymphoma cell line A20 and construct its eukaryotic expression vector pcDNA3.1-mCD99L2.</p><p><b>METHODS</b>The expression of mCD99L2 mRNA in A20 cell line was detected by in situ hybridization. The total RNA of A20 cells was extracted to obtain the full-length cDNA of the coding region of mCD99L2 gene by RT-PCR, the product of which was ligated into pMD18-T vector and the DNA sequence of the insert was detected. The coding regions of mCD99L2 gene was amplified from pMD-mCD99L2 by PCR using primers containing EcoR I and Xho I sites and cloned into the eukaryotic expression vector pcDNA3.1/MycHis(+).</p><p><b>RESULTS</b>In situ hybridization identified positive expression of mCD99L2 gene in the A20 cell line. The full-length cDNA of mCD99L2 coding region of A20 cell line was obtained by RT-PCR, which yielded a product of 712 bp as expected, and the DNA sequence was completely homologus to the mCD99L2 cDNA reported in GenBank. Restriction endonuclease digestion and DNA sequencing indicated that the eukaryotic expression vector pcDNA3.1(+)- mCD99L2 had been constructed successfully.</p><p><b>CONCLUSION</b>mCD99L2 cDNA has been cloned from mouse B lymphoma cell line A20 and its eukaryotic expression vector pcDNA3.1(+)- mCD99L2 successfully constructed, which facilitates further functional study of mCD99L2 gene in mouse B lymphoma cell line A20.</p>

Detection of bcl-2/IgH gene rearrangement in diffuse large B cell lymphoma by hemi-nested PCR / 中华血液学杂志

<p><b>OBJECTIVES</b>To explore a sensitive and specific method for detection of bcl-2/IgH gene rearrangement in diffuse large B cell lymphoma (DLBCL), and verify the credibility of the established method.</p><p><b>METHODS</b>bcl-2/IgH hemi-nested PCR primers were designed using the professional primer design software. Fifty-two samples of pathologically diagnosed DLBCL and 10 fresh tonsil tissues were amplified using hemi-nested touch down-PCR to detect bcl-2/IgH gene rearrangement. The PCR products were cloned and sequenced.</p><p><b>RESULTS</b>bcl-2/IgH gene rearrangement was detected in 6 of 52 DLBCL samples and 2 of 10 fresh tonsil tissues using one-way method. By using the hemi-nested PCR for the second round amplification, 5 of DLBCL were positive, but all of the fresh tonsil tissues were negative. The positive PCR products were sequenced and analyzed on the Internet, 3 of 8 cases obtained by one-way method were false positive, 5 positive cases amplified using hemi-nested PCR were all bcl-2/IgH gene rearrangement. PCR products of 3 false positive cases were homologous to BAC331191 and LLNLR-245D11 in human chromosome 19 and RP11-498P10 in chromosome 1.</p><p><b>CONCLUSION</b>There are false positive results using common primers for detecting bcl-2/IgH gene rearrangement. The mechanism may be that highly homologous sequences to human genome exist in commonly used primers. The specificity of the diagnosis could be improved by hemi-nested PCR using the combination of primers we designed and the traditional ones.</p>