A real world study on the relationship between drug resistance of targeted therapy and prognosis of HER-2-positive advanced breast cancer / 中华肿瘤杂志

Objective: To explore the effect of primary and acquired resistance to anti-human epidermal growth factor receptor 2 (HER-2) on the overall survival of patients with HER-2 positive advanced breast cancer. Methods: The clinical characteristics of HER-2 positive patients with advanced breast cancer admitted to Cancer Hospital of Chinese Academy of Medical Sciences from January 1998 to December 2018 were collected, and their neoadjuvant/adjuvant and advanced three-line chemotherapy were summarized. Among them, targeted drugs for HER-2 included trastuzumab, pertuzumab, T-DM1, RC48-ADC, lapatinib, pyrotinib, allitinib, sipatinib, seratinib. Based on the duration of benefit from anti HER-2 treatment, the patients were divided into two groups: primary anti HER-2 resistance group and acquired anti HER-2 resistance group. In this study, the overall survival (OS) was used as the main end point. Kaplan-Meier analysis and Cox proportional risk regression model were used to analyze the effects of different drug resistance mechanisms on the overall survival. Results: The whole group of 284 patients were included. The median age of recurrence and metastasis was 48 years old, 155 (54.6%) were hormone receptor (HR) positive and 129 (45.4%) were HR negative, 128 cases (45.1%) were premenopausal and 156 cases (54.9%) were postmenopausal, 277 cases (97.5%) had a score of 0-1 in ECoG PS and 7 cases (2.5%) had a score of more than 2 in the first diagnosis of relapse and metastasis. There were 103 cases (36.3%) in the primary drug resistance group and 181 cases (63.7%) in the secondary drug resistance group. The median overall survival time of the two groups was 24.9 months and 40.4 months, respectively, with statistical significance (P<0.001). Conclusion: Primary resistance to HER-2 is one of the factors of poor prognosis in HER-2 positive breast cancer, and its mechanism needs to be further explored.

Advances in bacterial Rieske non-heme iron ring-hydroxylating dioxygenases that initiate polycyclic aromatic hydrocarbons degradation / 生物工程学报

Polycyclic aromatic hydrocarbons (PAHs) are a class of persistent organic pollutants, which have received widespread attentions due to their carcinogenic and mutagenic toxicity. The microbial degradation of PAHs are usually started from the hydroxylation, followed by dehydrogenation, ring cleavage and step-by-step removal of branched chains, and finally mineralized by the tricarboxylic acid cycle. Rieske type non-heme iron aromatic ring-hydroxylating dioxygenases (RHOs) or cytochrome P450 oxidases are responsible for the conversion of hydrophobic PAHs into hydrophilic derivatives by the ring hydroxylation. The ring hydroxylation is the first step of PAHs degradation and also one of the rate-limiting steps. Here, we review the distribution, substrate specificity, and substrate recognition mechanisms of RHOs, along with some techniques and methods used for the research of RHOs and PAHs.

Real-time cell analysis combined with pattern recognition to evaluate anaphylaxis of Zushima Injection / 中国中药杂志

This paper was aimed to establish a new method for evaluating the anaphylactoid reaction of 15 batches of Zushima Injection from different manufacturers in vitro. Basophilic leukemia cell line RBL-2 H3 cells were cultured in vitro and Compound 48/80 was selected as positive drug. Real-time cell analysis(RTCA) system was used to detect the changes of cell index(CI) value after drug intervention. The degranulation of RBL-2 H3 cells was verified with the toluidine blue staining technology by observing the changes of cell morphology and skeleton. Clustering method was used to analyze the CI values of 15 batches of Zushima Injection on RBL-2 H3 cells. The results showed Compound 48/80(20 μg·mL~(-1)) significantly changed the cell morphology and cytoskeleton, with obvious degranulation. After adding Compound 48/80, CI value decreased rapidly within 30 minutes, then decreased slowly, suggesting that RTCA system can be used for rapid and sensitive evaluation of RBL-2 H3 cell degranulation. The results of cluster analysis showed that Zushima Injection from different manufacturers had different effects on RBL-2 H3 cells. S1-S8 and Compound 48/80 groups were grouped into one cluster, which suggesting that the sample might have potential clinical anaphylaxis. S9-S15 and the normal control group were grouped into one cluster, suggesting there was no anaphylactoid reaction in the sample. In this study, a rapid in vitro anaphylaxis evaluation technique based on RTCA system and pattern recognition method was established, which can be used for rapid in vitro evaluation of anaphylaxis for traditional Chinese medicine injection.

Optimal window setting for 40 keV virtual monoenergetic images derived from dual-layer spectral detector CT pancreas dynamic enhanced scanning / 中华放射学杂志

Objective:To explore the image quality of 40 keV virtual monoenergetic images (VMI) derived from dual-layer spectral detector CT （DLCT） pancreas dynamic enhanced scanning and its optimal window setting.Methods:From January to July 2019, 28 patients who underwent pancreas enhanced DLCT scan within one week before surgery and pathologically confirmed of pancreatic neuroendocrine tumors (pNETs) were retrospectively enrolled. Conventional polyenergetic images (PI) and 40 keV virtual monoenergetic images （VMI 40 keV） were generated after scanning.CT value of normal pancreatic parenchyma, lesion, abdominal subcutaneous fat, abdominal aorta and portal vein were measured in PI and VMI 40 keV. The contrast-to-noise ratio (CNR) of the pNETs lesion was calculated. All these objective results were compared between VMI 40 keV and PI using paired ttest. Individual window settings （W-Ind, including window width and window level） of VMI 40 keVwere recorded. Calculated window settings （W-Calc） were mathematically calculated via regression analysis and optimized window settings （W-Opt） were obtained.Subjective image quality was assessed with a 5-point scale and compared among VMI 40 keV with different window settings (W-Std, W-Ind, W-Calc and W-Opt) using Friedman test, and compared PI with standard abdominal window setting (W-Std) and VMI 40keV with W-Opt settings using Wilcoxon test.The maximum diameter of lesion was measured and compared with one-way ANOVA analysis among PI and VMI 40 keV with different windows settings. Results:For VMI 40 keV in both arterial phase and portal vein phase, the CT attenuation [(464.0±136.7), (375.4±79.2) HU] of pNETs lesion were statistically significantly higher than those in PI [（168.8±38.0), (140.5±23.5) HU] （ t=-16.107,-22.225, P<0.001）, CNR （16.5±11.1, 10.9±6.1） were also statistically significantly higher than those in PI （4.5±2.9, 3.0±1.9） （ t=-7.838, -9.781， P<0.001）,while with lower image noise in VMI 40 keV[（11.8±1.5),(11.8±1.4) HU] than PI （13.1±1.5,12.9±1.3 HU）（ t=6.356,3.891， P<0.001）. The subjective score for PI with W-Std and VMI 40 keV with W-Std, W-Ind, W-Calc, W-Opt in arterial phase were 4（1）, 1（0）, 5（0）, 5（0.75）, 5（1）, and which in portal vein phase were 3.5（1）, 1（0）, 5（0）, 5（0）, 5（1）.The subjective score of VMI 40 keV with different window settings had statistical differences （χ 2=76.143,76.000， P<0.001）. Compared to the image quality of PI with W-Std settings, VMI 40 keV with W-Opt settings have higher objective score （ Z=4.685, 4.235， P<0.001）. The maximum diameter of lesion has no statistical difference among PI and VMI 40 keV with different window settings （ F=0.008, 0.004， P>0.999） in both arterial phase and portal vein phase. Conclusions:The VMI 40 keV in pancreas dynamic enhancement scanning derived from dual-layer spectral detector CT have higher image quality than PI. Due to changes of CT value of tissue in VMI 40 keV, it is recommended to optimize window settings （window width/window level， 880/230 HU for arterial phase and 840/260 HU for portal vein phase） to obtain the best image quality.

Study of the optimal monochromatic image of coronary stent based on dual-layer detector spectral CT / 中华放射学杂志

Objective:To explore the optimal monochromatic level for the observation of coronary in-stent lumen by dual-layer spectral CT (DLCT).Methods:Forty-nine patients with 74 stents after percutaneous coronary intervention (PCI) who underwent coronary CTA (CCTA) examinations by a DLCT between January 2016 and September 2017 were retrospectively enrolled. A total of 12 groups of images including 60-120 keV (kilo electron voltage) images with 10 keV interval, 140-200 keV images with 20 keV interval and conventional images. In-stent lumen diameter of proximal, mid and distal portion was measured. Difference of CT values between in-stent lumen and ascending aorta was used to describe as blooming artifact, and noise of in-stent lumen as image noise. Then Likert 5-point scale was performed to evaluate images noise, enhancement of in-stent lumen, blooming artifact and diagnostic confidence. Differences of objective and subjective parameters among conventional and various monochromatic images were compared by Friedman test.Results:In the diameter measurement of the proximal, middle and distal segments of the stent, the difference between the images of each group was statistically significant (χ 2 = 427.270, 426.375, 400.981, P< 0.001). The diameter of the lumen measured by 120-200 keV single-level image was larger than that measured by 60-100 keV single-level image, and the difference was statistically significant ( P< 0.05). In the comparison of CT difference between the stent lumen and ascending aorta, the difference between the images of each group was statistically significant (χ 2 = 242.193, P< 0.001), and 100-200 keV single-level images were lower than the conventional images, the difference was statistically significant ( P< 0.05). In the comparison of noise values, the difference between the images of each group was statistically significant (χ 2 = 420.161, P< 0.001), and the difference was statistically significant ( P< 0.05). In the subjective scores of noise, enhancement, halo artifact and diagnostic confidence, there were statistically significant differences among the groups (χ2= 333.827, 455.989, 276.824, 399.497, P< 0.001). The noise score of 100-200 keV single-level image was higher than that of conventional image, the difference was statistically significant ( P< 0.05). The enhancement score of 60 keV was significantly higher than that of other images ( P< 0.05). The halo artifact score of 100-200 keV single level image was higher than that of 60-90 keV image, the difference was statistically significant ( P<0.05). The scores of 90-120 keV single-level images were higher than those of other single-level images, and the difference was statistically significant ( P< 0.05). Conclusions:CCTA examinations can be effectively performed by DLCT in patients after PCI in clinical settings, and 120 keV is recommended as the optimal monochromatic image for the observation of in-stent lumen.

Clinical usefulness of spectral images derived from dual-layer spectral detector CT for the detection of pancreatic neuroendocrine tumor lesions / 中华放射学杂志

Objective:To investigate the clinical value of iodine density map and low keV virtual monoenergetic images (VMI) derived from dual-layer spectral detector CT (DLCT) for the pancreatic neuroendocrine tumors (pNETs) detection.Methods:From January to June 2019, data of 23 pathologically confirmed patients of pNETs were retrospectively analyzed. All of the patients underwent pancreas enhanced DLCT scanning within 1 week before surgery. The conventional polyenergetic images (PI), iodine density map and 40, 50, 60, 70 keV VMI were generated. One resident radiologist with 3 years’ experience and one senior radiologist with over 10 years’ experience interpreted the images for the lesion detection independently using the following image series: PI, VMI (40-70 keV), PI combined with iodine density map. Lesion detection rates were recorded and compared among different image series. The CT value and noise of lesion, normal pancreatic parenchyma, and abdominal subcutaneous fat were measured in PI and VMI in both arterial and portal vein phases. The contrast-to-noise ratio (CNR) of lesion was calculated. The CT value of lesion and normal pancreatic parenchyma, CNR of lesion, and image noise were compared using repeated one-way ANOVA test. Subjective image quality was assessed with a 5-point scale and compared with Friedman test.Results:A total of 26 lesions were confirmed from 23 patients. For resident radiologist and senior radiologist, the detection rates of pNETs lesion using PI were 76.9% (20/26) and 84.6% (22/26) respectively, and both improved to 92.3% (24/26) using image series of 40 and 50 keV VMI. For senior radiologist, the pNETs lesion detection rate was further improved to 96.2% (25/26) using image series of PI with iodine map. The CT value of lesion and normal pancreatic parenchyma, CNR, and image noise had statistical differences among PI and VMI (40-70 keV) in both arterial and portal vein phase ( P<0.001). The mean CT attenuation and CNR of lesion in VMI increased significantly as the energy level decreased.The CNR of lesion in all VMI (40-70 keV) was significantly higher than that in PI. The median of subjective scores of image quality in PI and VMI (40-70 keV) were 3, 3, 4, 4, and 5 respectively, and the difference was statistically significant (χ2=66.393, P<0.001). Conclusions:The low keV VMI derived from DLCT can increase the CT value and CNR of pNETs, and the lesion detection rate can be improved combined with iodine density map. The CNR of pNETs is the highest in 40 keV VMI, and image noise is still lower than that of PI, so 40 keV VMI is recommended for clinical application.

Proteomic analysis and verification of protein expression after upregulation of human CD99 in Hodgkin lymphoma cell line L428 / 中华血液学杂志

Objective: To investigate the proteins expression difference after upregulation of human CD99 in Hodgkin Lymphoma cell line, L428 cell, and verify the function of differential proteins. Methods: The differential proteins were detected by two-dimensional fluorescence difference gel electrophoresis and mass spectrometry analysis, cluster analysis was done by GOfact. Results: There were 38 proteins screened out, of which 21 proteins were positively associated with CD99, while 17 proteins were negative. Among the 38 proteins, 32 proteins participated in biological process, and 35 proteins were involved in the composition and construction. And 28 proteins participated in multifaceted biological activities including antioxidation, protein binding, catalytic activity, regulation of enzyme, signal transduction, molecular structure, regulation of translation and ion transport. Conclusions: The changes of the differential proteins, correlated with cytoskeleton, cell differentiation, signal pathway and regulating gene expression, are closely relevant to the translation between Hodgkin/Reed-Sternberg and B lymphocyte cell.

Feasibility study on dual-layer spectral detector CT-derived virtual non-contrast images substitute for true non-contrast images / 中华放射学杂志

Objective To explore the feasibility of the virtual non-contrast images derived from dual-layer spectral detector CT (SDCT) substitute for true non-contrast images.Methods From July 2017 to September 2017,40 patients under-went pre-and arterial-venous dual-phase post-contrast abdominal imagining on a SDCT in Shanghai Jiaotong University School of Medicine Affiliated Ruijin Hospital.The images were retrospectively analyzed.The arterial VNC images (VNC-a) and venous VNC images (VNC-v)were derived from spectral based image (SBI) datasets of arterial and portal venous phase using a dedicated software respectively.Then mean CT attenuation,mean image noise,signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were compared with one-way ANOVA analysis among TNC,VNC-a,VNC-v images in liver,spleen,pancreas,abdominal aorta,inferior vena cava,psoas muscle,L4 vertebra and perirenal fat.And Bland-Altman plots was used to analyze the CT attenuation equivalence between TNC and VNC.Subjective image quality was assessed with a 5-point scale and compared with Friedman H test.The dose length product (DLP) of pre-and post-contrast scans were recorded.Results The CT attenuation of abdominal aorta,perirenal fat,L4.vertebra among three kinds of images had significant differences (P＜0.05)which overestimated the CT attenuation of perirenal fat and underestimated CT attenuation of abdominal aorta and vertebra compared to TNC,VNC.Except for pancreas,the image noise of all other tissues among three kinds of images had significant differences,images noise of VNC images were lower than TNC images (P＜0.05).The SNR of liver,spleen,psoas muscle and inferior vena cava and CNR of pancreas,abdominal aorta,inferior vena cava,L4 vertebra and perirenal fat were statistically significant among the three images (P＜0.05).Except for vertebra and perirenal fat,good equivalence between VNC and TNC images were observed in all relevant tissues with Bland-Altman analysis.Image quality subjective scoring of TNC,VNC-a,and VNC-v were 5.00(1.00),5.00(0.75),5.00(1.00) respectively which had no significant differences (P=0.20).The DLP of TNC,arterial and venous phase scan were (255.2±62.0),(258.9±62.9),(252.0±61.2)mGy·cm,and the total DLP of dual-phase contrast enhanced scan was (766.2± 185.3) mGy· cm.The DLP of TNC accounted for 33.3％(255.2/766.2) of the total DLP,and effective dose of TNC was (3.83±0.90)mSv.Conclusions CT attenuation in VNC image which exhibited lower image noise was identical to TNC images in the majority of abdomen tissues except for vertebra and perirenal fat.and the VNC image.The VNC image which reduces radiation dosage derives from SDCT enhanced image might be used as a substitute for the TNC image.

Advanced glycation end products promote differentiation of CD4(+) T helper cells toward pro-inflammatory response / 华中科技大学学报（医学）（英德文版）

This study investigated the effect of advanced glycation end products (AGEs) on differentiation of naïve CD4(+) T cells and the role of the receptor of AGEs (RAGE) and peroxisome proliferator-activated receptors (PPARs) activity in the process in order to gain insight into the mechanism of immunological disorders in diabetes. AGEs were prepared by the reaction of bovine serum albumin (BSA) with glucose. Human naïve CD4(+) T cells, enriched from blood of healthy adult volunteers with negative selection assay, were cultured in vitro and treated with various agents including AGEs, BSA, high glucose, PGJ2 and PD68235 for indicated time. In short hairpin (sh) RNA knock-down experiment, naïve CD4(+) T cells were transduced with media containing shRNA-lentivirus generated from lentiviral packaging cell line, Lent-X(TM) 293 T cells. Surface and intracellular cytokine stainings were used for examination of CD4(+) T cell phenotypes, and real-time PCR and Western blotting for detection of transcription factor mRNA and protein expression, respectively. The suppressive function of regulatory T (Treg) cells was determined by a [(3)H]-thymidine incorporation assay. The results showed that AGEs induced higher pro-inflammatory Th1/Th17 cells differentiated from naïve CD4(+) T cells than the controls, whereas did not affect anti-inflammatory Treg cells. However, AGEs eliminated suppressive function of Treg cells. In addition, AGEs increased RAGE mRNA expression in naïve CD4(+) T cells, and RAGE knock-down by shRNA eliminated the effect of AGEs on the differentiation of CD4(+) T cells and the reduction of suppressive function of Treg cells. Furthermore, AGEs inhibited the mRNA expression of PPARγ, not PPARα PPARγ agonist, PGJ2, inhibited the effect of AGEs on naïve CD4(+) T cell differentiation and reversed the AGE-reduced suppressive function of Treg cells; on the other hand, PPARγ antagonist, PD68235, attenuated the blocking effect of RAGE shRNA on the role of AGEs. It was concluded that AGEs may promote CD4(+) T cells development toward pro-inflammatory state, which is associated with increased RAGE mRNA expression and reduced PPARγ activity.

Advanced glycation end products promote differentiation of CD4(+) T helper cells toward pro-inflammatory response / 华中科技大学学报（医学）（英德文版）

This study investigated the effect of advanced glycation end products (AGEs) on differentiation of naïve CD4(+) T cells and the role of the receptor of AGEs (RAGE) and peroxisome proliferator-activated receptors (PPARs) activity in the process in order to gain insight into the mechanism of immunological disorders in diabetes. AGEs were prepared by the reaction of bovine serum albumin (BSA) with glucose. Human naïve CD4(+) T cells, enriched from blood of healthy adult volunteers with negative selection assay, were cultured in vitro and treated with various agents including AGEs, BSA, high glucose, PGJ2 and PD68235 for indicated time. In short hairpin (sh) RNA knock-down experiment, naïve CD4(+) T cells were transduced with media containing shRNA-lentivirus generated from lentiviral packaging cell line, Lent-X(TM) 293 T cells. Surface and intracellular cytokine stainings were used for examination of CD4(+) T cell phenotypes, and real-time PCR and Western blotting for detection of transcription factor mRNA and protein expression, respectively. The suppressive function of regulatory T (Treg) cells was determined by a [(3)H]-thymidine incorporation assay. The results showed that AGEs induced higher pro-inflammatory Th1/Th17 cells differentiated from naïve CD4(+) T cells than the controls, whereas did not affect anti-inflammatory Treg cells. However, AGEs eliminated suppressive function of Treg cells. In addition, AGEs increased RAGE mRNA expression in naïve CD4(+) T cells, and RAGE knock-down by shRNA eliminated the effect of AGEs on the differentiation of CD4(+) T cells and the reduction of suppressive function of Treg cells. Furthermore, AGEs inhibited the mRNA expression of PPARγ, not PPARα PPARγ agonist, PGJ2, inhibited the effect of AGEs on naïve CD4(+) T cell differentiation and reversed the AGE-reduced suppressive function of Treg cells; on the other hand, PPARγ antagonist, PD68235, attenuated the blocking effect of RAGE shRNA on the role of AGEs. It was concluded that AGEs may promote CD4(+) T cells development toward pro-inflammatory state, which is associated with increased RAGE mRNA expression and reduced PPARγ activity.

Expression of miR-9 in B lymphocytes and B cell lymphomas cell lines and its significance / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of miR-9 in B lymphocytes, B cell lymphoma and classical Hodgkin's lymphoma (cHL) cell lines and its significance.</p><p><b>METHODS</b>CD19(+) B lymphocytes were sorted from normal lymph node by magnetic beads. Total cellular micro-RNA was extracted from cHL cell line L428, B cell lymphoma cell lines Ly1 and Ly10 (diffuse large B cell lymphoma), Raji cells (Burkitt's lymphoma) and CD19(+) B lymphocytes, respectively. These micro-RNAs were separately transformed into cDNA by reverse transcription. The expression levels of miR-9 were measured by fluorescence quantitative PCR. In situ hybridization was used to detect the expression of miR-9 in cell lines.</p><p><b>RESULTS</b>The expression of miR-9 was high in L428 cells (104.44 ± 1.61), and low in cell lines of B cell lymphoma (Ly1: 2.17 ± 0.38; Ly10: 1 ± 0.015; Raji: 2.65 ± 0.89), and extremely low in CD19(+) B lymphocytes (0.0026 ± 0.00040). Compared with that in the other cell lines, the expression of miR-9 in L428 cells was statistically significant (P < 0.05). miR-9 localized in the cytoplasm diffusely and strongly in L428, but scattered and slightly with some prominent distribution around the nuclear membranes in Ly1 and Ly10, and only weakly in Raji.</p><p><b>CONCLUSIONS</b>miR-9 highly expressed in cHL cell line and might be a molecular marker for diagnosis and treatment of cHL.</p>

Preparation of RNA probe for cd99l2 gene of zebrafish labeled with digoxingenin-UTP / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the expression pattern of cd99l2 gene during zebrafish development, the RNA probes for whole-mount in situ hybridization were prepared in this study.</p><p><b>METHODS</b>The cd99l2 fragment obtained by RT-PCR was cloned into pGM-T Easy, then the plasmids were linearized with the restriction enzymes SacII or SalI. Using Sp6 or T(7) RNA polymerase, the digoxingenin-labeled antisense and sense probes were synthesized and confirmed by whole-mount in situ hybridization.</p><p><b>RESULTS</b>The plasmid cd99l2/pGM-T was constructed. cd99l2 gene expression pattern during embryogenesis of zebrafish was examined using the antisense probe, and intense expression was detected in the central nervous system during zebrafish development.</p><p><b>CONCLUSION</b>The antisense probe can be used for study of the spatial and temporal distribution of cd99l2 during zebrafish development using the sense probe as control.</p>

Primers for detecting gene rearrangement in different regions of immunoglobulin heavy chain genes and their application in diagnosis of paraffin-embedded lymphoma tissues / 南方医科大学学报

<p><b>OBJECTIVE</b>To analyze and optimize the gene rearrangement primers of different frame regions (FR) of immunoglobulin heavy chain (IgH) genes by bioinformatic methods and explore the application of these primers in the detection of paraffin-embedded lymphoma tissues.</p><p><b>METHODS</b>Three pairs of primers from IgH FR1, FR2 and FR3 regions (P1c, P2A and P31, respectively) were selected as the B cell gene rearrangement primers after comparison of the gene fragments in 44 IgH variable and 6 joining regions. Using one pair of T cell receptor (TCR) gamma primer as the T cell gene rearrangement primer, 101 histopathologically confirmed lymphoproliferative samples including 80 B cell lymphomas, 14 T cell lymphomas, and 7 reactive proliferative lymph nodes were examined by PCR for gene arrangement. The DNAs from DG75 and Jurkat cell lines were used as the positive controls for B and T cell lymphoma, respectively, with those from reactive proliferative lymph nodes as the negative control.</p><p><b>RESULTS</b>The positivity rates of IgH primers (P1c, P2A and P31) in the 80 B cell lymphomas were 37.5% (30/80), 52.5% (42/80) and 70.0% (56/80), respectively, and only one of the 14 T cell lymphoma cases was positive for the primers, suggesting significant differences in the detection rates of B cell lymphomas by the 3 primers. The detection rate was increased to 83.9% by combining the results by P31 and P2A primers. No positivity was found in the proliferative reaction tissues.</p><p><b>CONCLUSION</b>Primers from IgH FR3 region genes are more sensitive than that from the FR1 and FR2 regions in the detection of gene rearrangement in paraffin-embedded lymphoma tissues. The detection rates can be increased by combining the results with the primers for IgH FR3 with that of FR2.</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>

Single plane Simpson's method for measurement of right ventricular volume by X-ray ventriculography / 中华心血管病杂志

<p><b>OBJECTIVE</b>To study the validity of single plane Simpson's method with conventional X-ray ventriculography for estimation of right ventricular (RV) volume.</p><p><b>METHODS</b>Fifteen human RV casts were obtained from 15 subjects who died from non-cardiac causes within 24 hours after death. These casts were photographed respectively and their volumes were calculated by using the single plane Simpson's method based on a new half-circle model. The actual RV cast volumes were determined by water displacement method.</p><p><b>RESULTS</b>The actual RV volume was (64.23 +/- 24.51) ml and the calculated volume was (58.04 +/- 24.45) ml. The calculated RV volume underestimated the actual volume by (6.19 +/- 12.38) ml, but there was no significant difference between the actual and the calculated RV volume (P > 0.05). There was a significant correlation between the actual cast volume and the calculated volume (r = 0.983, P < 0.01). The regression equation was: RV actual volume = 1.074 x (RV calculated volume).</p><p><b>CONCLUSION</b>RV volume calculated by single plane Simpson's method with conventional X-ray ventriculography is accurate and deserves further study.</p>

Unspecified peripheral T cell lymphoma with distinct lymphoid follicules / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the morphological features and immunophenotype of unspecified peripheral T cell lymphoma with distinct lymphoid follicular growth pattern.</p><p><b>METHODS</b>Three cases of peripheral T cell lymphoma with special pathohistological features were collected. Morphologic analysis and immunohistochemical staining for CD3, CD45RO, CD43, CD20, CD79a, cyclinD1, bcl-2, CD4, CD8 and S-100 were performed. PCR was used to study TCR gamma gene rearrangements.</p><p><b>RESULTS</b>The main symptoms of all the three patients with the primary sites of cervix and lower jaw. There were intermittent fever and skin rashes in the course of the disease. Morphological study showed lymphoid follicular reactive hyperplasia, mantle zone disappear, prominent infiltration of marginal zones by medium-sized tumor cells with clear cytoplasm and significant nuclear atypia. The immunophenotypic profile confirmed that they were T cell lymphomas. TCR gamma gene rearrangements were found in all the three patients.</p><p><b>CONCLUSION</b>In some unspecified peripheral T cell lymphomas, the distinct follicular growth pattern and incomplete effacement of the lymph node architecture make it necessary to differentiate them from reactive hyperplasia, marginal zone B cell lymphoma, follicular B cell lymphoma and mantle cell lymphoma.</p>

Correcting the congenital ala defect of the facial cleft with revolving reposition / 中华整形外科杂志

<p><b>OBJECTIVE</b>To introduce the experience of correcting the congenital ala defect of facial cleft with the method of revolving reposition.</p><p><b>METHODS</b>Five patients were treated with this method. Based on features of the defect, two different operations were employed with the medial crura or the lateral crura of the ala as the revolving pivot.</p><p><b>RESULTS</b>All the operations obtained good results.</p><p><b>CONCLUSION</b>The operation of revolving reposition for congenital ala defect is a simple practical method with satisfactory results.</p>

Non-replicating recombinant vaccinia virus expressing HPV16 E6 and E7 proteins elicits anti-tumor immunity in mice / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the anti-tumor immunity of the non-replicating recombinant vaccinia virus expressing HPV16 E6 and E7 proteins.</p><p><b>METHODS</b>C57BL/6 mice were immunized by non-replicating recombinant vaccinia virus (NTVJmE6E7), and then specific CTLs were determined. Immune protection effects were evaluated by challenges of different doses of TC-1 tumor cells. Immunotherapeutic effects in form of recurrence were evaluated on the tumor-removed mice.</p><p><b>RESULTS</b>Mice immunized by NTVJmE6E7 could generate TC-1 cell specific cytotoxic T lymphocyte (CTL). Mice boosted with NTVJmE6E7 could tolerate the challenge of 1 x 10(4) TC-1 cells. NTVJmE6E7 could effectively prevent the tumor recurrence in the tumor-removed mice.</p><p><b>CONCLUSION</b>NTVJmE6E7 can be taken as a candidate of therapeutic vaccine for HPV-associated tumors and their precursor lesions.</p>