Effects of Notch-1 down-regulation on malignant behaviors of breast cancer stem cells / 华中科技大学学报（医学）（英德文版）

This study examined the effect of Notch-1 signaling on malignant behaviors of breast cancer cells by regulating breast cancer stem cells (BCSCs). BCSCs were enriched by using serum-free medium and knocked out of Notch-1 by using a lentiviral vector. Real-time polymerase chain reaction (RT-PCR) and Western blotting were used to detect the Notch-1 expression levels in breast cancer cell lines and BCSCs, and flow cytometry to detect the proportion of BCSCs in BCSC spheres. The BCSC self-renewal, migration, invasion, and tumorigenicity were examined by the tumor microsphere-forming assay and transwell assay and after xenotransplantation. The results showed that the Notch-1 silencing reduced the number of BCSC spheres, the proportion of BCSCs, and the number of cells penetrating through the transwell membrane. It also decreased the size of tumors that were implanted in the nude mice. These results suggest that Notch-1 signaling is intimately linked to the behaviors of BCSCs. Blocking Notch-1 signaling can inhibit the malignant behaviors of BCSCs, which may provide a promising therapeutical approach for breast cancer.

Effects of Notch-1 down-regulation on malignant behaviors of breast cancer stem cells / 华中科技大学学报（医学）（英德文版）

This study examined the effect of Notch-1 signaling on malignant behaviors of breast cancer cells by regulating breast cancer stem cells (BCSCs). BCSCs were enriched by using serum-free medium and knocked out of Notch-1 by using a lentiviral vector. Real-time polymerase chain reaction (RT-PCR) and Western blotting were used to detect the Notch-1 expression levels in breast cancer cell lines and BCSCs, and flow cytometry to detect the proportion of BCSCs in BCSC spheres. The BCSC self-renewal, migration, invasion, and tumorigenicity were examined by the tumor microsphere-forming assay and transwell assay and after xenotransplantation. The results showed that the Notch-1 silencing reduced the number of BCSC spheres, the proportion of BCSCs, and the number of cells penetrating through the transwell membrane. It also decreased the size of tumors that were implanted in the nude mice. These results suggest that Notch-1 signaling is intimately linked to the behaviors of BCSCs. Blocking Notch-1 signaling can inhibit the malignant behaviors of BCSCs, which may provide a promising therapeutical approach for breast cancer.

Accurate assessment of HER2 gene status for invasive component of breast cancer by combination of immunohistochemistry and chromogenic In Situ hybridization / 华中科技大学学报（医学）（英德文版）

The specimens of ductal carcinoma in situ (DCIS) with early invasion, and specimens collected by core needle biopsy (CNB) tend to contain limited amount of invasive component, so it is imperative to explore a new technique which can assess HER2 gene status accurately for the limited invasive cancer component in these specimens. Dual staining technique of combining immunohistochemistry (IHC) for myoepithelial cells and single or dual probe chromogenic in situ hybridization (CISH) for HER2 gene was performed on routinely processed paraffin sections from 20 cases diagnosed as having DCIS with invasive cancer. Among them, 10 had fluorescence in situ hybridization (FISH)-confirmed amplification of HER2 and 10 had FISH-confirmed non-amplification of HER2. We successfully detected HER2 genetic signals and myoepithelial IHC markers (SMM-HC or CK5/6) simultaneously on a single section in all 20 specimens. Myoepithelial markers and HER2 signals detected by dual staining assay were consistent with those by individual technique performed alone. HER2 gene amplification results determined by dual staining assay were 100% consistent with those of FISH. Dual staining technique which allows simultaneous detection of myoepithelial marker protein and cancerous HER2 gene is feasible, and it has potential to be used in clinical practice for effective determination of HER2 amplification in limited invasive component.

Accurate assessment of HER2 gene status for invasive component of breast cancer by combination of immunohistochemistry and chromogenic In Situ hybridization / 华中科技大学学报（医学）（英德文版）

The specimens of ductal carcinoma in situ (DCIS) with early invasion, and specimens collected by core needle biopsy (CNB) tend to contain limited amount of invasive component, so it is imperative to explore a new technique which can assess HER2 gene status accurately for the limited invasive cancer component in these specimens. Dual staining technique of combining immunohistochemistry (IHC) for myoepithelial cells and single or dual probe chromogenic in situ hybridization (CISH) for HER2 gene was performed on routinely processed paraffin sections from 20 cases diagnosed as having DCIS with invasive cancer. Among them, 10 had fluorescence in situ hybridization (FISH)-confirmed amplification of HER2 and 10 had FISH-confirmed non-amplification of HER2. We successfully detected HER2 genetic signals and myoepithelial IHC markers (SMM-HC or CK5/6) simultaneously on a single section in all 20 specimens. Myoepithelial markers and HER2 signals detected by dual staining assay were consistent with those by individual technique performed alone. HER2 gene amplification results determined by dual staining assay were 100% consistent with those of FISH. Dual staining technique which allows simultaneous detection of myoepithelial marker protein and cancerous HER2 gene is feasible, and it has potential to be used in clinical practice for effective determination of HER2 amplification in limited invasive component.

Pulmonary embolism excluded acute coronary syndrome by coronarography: a retrospective analysis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Pulmonary embolism (PE) is often mistaken as acute coronary syndromes (ACS) because of the considerable overlap in their clinical features. We evaluated the factors causing misdiagnosis of PE as ACS and factors that differentiate PE from ACS to improve the diagnosis efficacy of PE.</p><p><b>METHODS</b>The medical records of 22 consecutive PE patients, between 2001 and 2010, who were initially suspected of ACS were retrieved. ACS was ruled out by coronary artery angiography before a definite diagnosis of PE was given. Twenty-two contemporary cases of ACS matched by age and sex were recruited as controls. Clinical manifestations, electrocardiograms (ECG), and biomarkers of these patients were reviewed retrospectively. The factors causing misdiagnosis of PE as ACS and factors differentiating PE from ACS were evaluated.</p><p><b>RESULTS</b>We found two leading causes of misdiagnosis of PE as ACS. One is that PE can resemble ACS in several clinical aspects (symptoms and signs, ECG findings, plasma cardiac troponin I, and D-dimer). The other is the insufficient recognition of PE by clinicians. Risk factors for venous thromboembolism (VTE), especially deep venous thrombosis (DVT), together with signs of PE, such as unexplained dyspnea or hypoxemia, and right ventricular pressure overload on ECGs are valuable in differentiating the two diseases.</p><p><b>CONCLUSIONS</b>Differentiation between PE and ACS is sometimes challenging. Adequate awareness of the risk factors for VTE and the signs of PE are crucial in the diagnosis of PE.</p>

Depression, anxiety and influencing factors in patients with acute pulmonary embolism / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Psychological distress has been widely studied in many cardiovascular and pulmonary diseases, but the condition in acute pulmonary embolism (APE) is unknown. The purpose of this study was to investigate levels of depression and anxiety and their influencing factors in APE patients.</p><p><b>METHODS</b>Sixty consecutive patients with APE were subjected to investigation of depression and anxiety by the Beck Depression Inventory and State-Trait Anxiety Inventory, and 60 community-based subjects were enrolled as controls. APE patients were stratified as high-risk, intermediate-risk and low-risk according to the disease severity. Scores of depression and anxiety were compared by statistical analysis using paired t tests between APE patients and controls, and by analysis of variance within the APE patients with the three risk stratification. Factors influencing depression and anxiety were evaluated.</p><p><b>RESULTS</b>The mean age of the patients (38 males and 22 females) was (52 ± 12) years. APE patients displayed higher scores of depression (P = 0.04) and anxiety (P = 0.001) compared with controls. Patients in the high-risk group displayed higher scores of depression (P = 0.004) and anxiety (P = 0.001) compared with those in the intermediate- and low-risk groups. Depression scores were highly correlated with anxiety scores (r = 0.60, P < 0.001). Both depression and anxiety inversely related to risk stratification (P < 0.01), age (P < 0.05), and arterial blood oxygen pressure (PaO2) (P < 0.05). Linear regression analysis showed that PaO2 was independently inversely related to both depression (P < 0.01) and anxiety (P < 0.05); risk stratification and age were independently inversely related to anxiety (P < 0.05).</p><p><b>CONCLUSIONS</b>Patients of APE suffered high levels of depression and anxiety, which were negatively influenced by PaO2, risk stratification and age.</p>

Operation for differentiated thyroid cancer: a experience of 546 cases / 中华外科杂志

<p><b>OBJECTIVE</b>To approach reasonable operational mode of differentiated thyroid cancer.</p><p><b>METHODS</b>Retrospectively review 546 differentiated thyroid cancer patients who received bilateral thyroidectomy with or without cervical lymph node excision from January 2001 to December 2006.</p><p><b>RESULTS</b>No death case happened during operation and hospitalization. The positive percentage of cervical lymph node metastasis was 76.2% (358/470). The incidence rate of single lateral recurrent laryngeal nerve injury was 1.1% (6 cases), parathyroid gland partly injury was 0.4% (2 cases), superior laryngeal nerve injury was 0.7% (4 cases), bleeding was 0.6% (3 cases) and esophagus injury after operation was 0.2% (1 case). There were no bilateral recurrent laryngeal nerve injury found.</p><p><b>CONCLUSIONS</b>Total thyroidectomy is an essential operational mode for differentiated thyroid cancer. It is necessary to excise cervical lymph node when the tumor's diameter exceeds 1 cm.</p>

Pathological manifestation and feasibility of conservative breast surgery after effective neoadjuvant chemotherapy for breast cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the pathological manifestation of the lesion as well as the feasibility of conservative breast surgery after neoadjuvant chemotherapy of patients with locally advanced breast cancer.</p><p><b>METHODS</b>Clinic data of the 46 patients treated by neoadjuvant chemotherapy effectively and surgical procedures from June 2003 to August 2007 were analyzed. All the patients were female, aged from 28 to 65 years old with a mean of 46.3 years old. The diameter of neoplasm was 7 to 13 cm. All the patients had been treated by neoadjuvant chemotherapy effectively and received modified radical mastectomy or Halsted radical mastectomy. The specimen slices were cut from 6 different directions taking the residual cancerous lesion as the center.</p><p><b>RESULTS</b>Nineteen patients (41.3%) were concentrating contracted, with complete remission in 6 patients (31.6%) and residual focus in peripheral zone in 13 patients (68.4%). Twenty-seven patients (58.7%) responded in a sifter-like pattern.</p><p><b>CONCLUSION</b>For most patients with locally advanced breast cancer treated by neoadjuvant chemotherapy effectively, their pathologic features are not up to the conservative breast surgery criterion, conservative breast surgery should be cautiously considered.</p>

Outcome of reoperation for thyroid cancer: clinical analysis of 72 cases / 中华外科杂志

<p><b>OBJECTIVE</b>To analyze the indication of reoperation of thyroid cancer and to explore the timing, surgical pattern of reoperation.</p><p><b>METHODS</b>Protocols of 72 patients underwent reoperation of thyroid cancer from June 2003 to August 2006 were reviewed retrospectively. Causes for reoperation were as follows: residue of the tumor locally as the inappropriate initial operation; local recurrence and cervical lymph node metastasis; before (131)I ablation which differentiated thyroid cancer with distant place metastasis. The reoperation style included residual lobectomy plus isthmus with single tumor below 2 cm, total thyroidectomy in most the other conditions and selective lymph node dissection in finding or suspected cervical lymph node metastasis.</p><p><b>RESULTS</b>The rate of residual in thyroid confirmed by postoperative pathology was 47.1% (32/68). The rate of residual in cervical lymph node was 81.4% (35/43). The rates of temporary and permanent laryngeal recurrent nerve injury were 5.6% (4/72) and 1.4% (1/72) respectively. The rates of temporary and permanent hypocalcemia were 26.4% (19/72) and 1.4% (1/72) respectively.</p><p><b>CONCLUSIONS</b>Inadequate operation, local recurrence and cervical lymph node metastasis of thyroid cancer need revision surgery undoubtedly. The optimal treatment was total thyroidectomy and level VI, VII central compartment lymph node dissection plus intraoperative frozen-section evaluation.</p>

Effect of PC-cell derived growth factor shRNA on estrogen dependent of estrogen receptor negative breast cancer cell lines / 中华外科杂志

<p><b>OBJECTIVE</b>To analyze the relationship between estrogen dependence of breast cancer cells and expression level of PC-cell derived growth factor (PCDGF) and investigate the possibility of practicing endocrine therapy in estrogen receptor negative breast cancer.</p><p><b>METHODS</b>Expression level of PCDGF mRNA was detected by fluorescence quantitative polymerase chain reaction and cell growth curve was drawn by cell count kit-8 method, then analyzed the relationship between the expression of PCDGF and dependence of estrogen. PCDGF shRNA vector was constructed and transfected into breast cancer cell lines (MCF-7 and MDA-MB-231) using Lipofectamin 2000, examined and compared the changes of estrogen dependence between the two cell lines.</p><p><b>RESULTS</b>PCDGF was expressed in most breast cancer cell lines, and the growth dependence of estrogen was higher in the cells with high-expressing PCDGF than those with low-expressing PCDGF. Expression of PCDGF could be inhibited by transfected shRNA into MCF-7 and MDA-MB-231 (inhibition rates were 81.1% and 86.7% respectively), inhibition of the expression of PCDGF could lead to higher growth dependence of estrogen, and the dependence of MDA-MB-231 was increased much more than MCF-7(P < 0.05).</p><p><b>CONCLUSIONS</b>The expression level of PCDGF is higher in estrogen receptor negative breast cancer than in estrogen receptor positive breast cancer. Inhibiting the expression of PCDGF could reverse resistance of endocrine therapy especially to estrogen receptor negative breast cancer.</p>

The chemotherapy role of vinorelbine with relapsed breast cancer primary cells in vitro / 肿瘤研究与临床

Objective To know the antitumor effect and value of vinorelbine in chemotherapy with relapsed breast cancer.Methods To receive breast cancer tissue from primary and relapsed patients by surgery,then to obtain mammary cancer cells by collagenase.The antitumor effect of 5 chemotherapy drugs with breast cancer cells was measured by culture in vitro and MTT.Results With primary cancer cells from primary breast cancer,the antitumor effect of paclitaxel and docetaxel(sensitivity is 91.04%,92.54% respec- tively) is better than adriamycin,epirubicin and gemcitabine(sensitivity is 73.13%,74.63%,71.64% re- spectively;P

Cloning and Expression of EGFR Tyrosine Kinase and Construction of the Screening Model for the Enzymatic Inhibitors / 中国生物工程杂志

As a member of tyrosine kinases(TK) family,Epidermal growth factor receptor(EGFR) has an activity of intrinsic protein tyrosine kinase,which plays an essential role in the regulation of signal transduction in the cells.Due to the abnormal expression of EGFR-TK,the certain type cancers may developed and progressed.Based on that,the inhibitors of EGFR-TK could be effective medicines for the treatment of cancer.The EGFR-TK domain was amplified by RT-PCR with RNA of HUVCEs cells as the template and expressed in E.coliBL21(DE3) using plasmid pET30a as vector.The recombinant protein was purified with the affinity chromatography(Ni-NTA),which was identified to have kinase activity catalyzing the substrate phosphorylated with ATP in the enzymatic reaction.Using the recombinant EGFR-TK as target,the screening model for enzymatic inhibitors was constructed.