Flapless one-knot suture: a novel surgical technique for transscleral fixation of C-loop intraocular lenses.

AIM: To describe a novel suture approach for transscleral fixation of C-loop intraocular lenses (IOL) and to compare the surgical outcomes with the four-haptics posterior chamber (PC)-IOL technique. METHODS: We retrospectively analyzed 16 eyes of 16 patients who underwent transscleral fixation of C-loop PC-IOLs using a flapless one-knot suture technique, which were followed up for longer than 17mo. In this technique, the capsulorless IOL was suspended using a single suture for transscleral fixation of four feet. Then we compared its surgical outcomes and complications with the four-haptics PC-IOLs using the Student's t test and Chi-square test. RESULTS: Sixteen patients of 16 eyes with a mean age of 58.3±10.1y (42-76y) who received transscleral C-loop IOL implantation due to trauma, vitrectomy, or cataract surgery with inadequate capsule support showed improved visual acuity. The difference was not significant between two IOLs except the surgery time (P>0.05). The mean operation times of C-loop IOL surgery was 24.1±1.83min and 31.3±4.47min of the four-haptics PC-IOL method (P<0.0001). In the C-loop IOLs group, there was statistical difference between the preoperative and the postoperative UCVA (logMAR, 1.20±0.50 vs 0.57±0.32, P=0.0003). There was no statistical difference between the preoperative and the postoperative BCVA (logMAR, 0.66±0.46 vs 0.40±0.23, P=0.056). However, there was no statistically significant difference in postoperative UCVA and BCVA between the two IOLs (P>0.05). We did not detect any optic capture, IOL decentration or dislocation, suture exposed, or cystoid macular edema in patients underwent C-loop IOLs surgery. CONCLUSION: The novel flapless one-knot suture technique for transscleral fixation of C-loop IOL is a simple, reliable, and stable technique.

Screening specific polypeptides of breast cancer stem cells from a phage display random peptide library.

The present study aimed to identify polypeptides that specifically bond to breast cancer stem cells from a phage display random 12 peptide library, in addition to the affinity and specificity of polypeptides. A phage display random 12 peptide library was screened using breast cancer stem cells as targets isolated from the MDA-MB-231 cell line using the serum-free culture technique with hs578bst and MDA-MB-231 cells as subtract-screening cells. Positive and specific binding clones were amplified and sent for sequencing. The affinity and specificity of the positive clones were subsequently identified by ELISA and 3,3'-diaminobenzidine staining. The results demonstrated that phages were gathered ~500 times following three rounds of biopanning. ELISA identified that the affinity to breast cancer stem cells of the no. 6 phage was 6.14 times higher than that in the control group. In addition, immunohistochemistry observed that the no. 6 phage exhibited high-specificity bonding to breast cancer stem cells, and the peptide sequence of the positive phage was GYSASRSTIPGK following DNA sequencing and translation. Thus, the present study isolated a specific peptide that bonds to breast cancer stem cells from a phage display random peptide library, which may facilitate further studies regarding the stem cell-targeted therapy of breast cancer.

Antisense oligodeoxynucleotide against human telomerase reverse transcriptase inhibits the proliferation of Eca-109 esophageal carcinoma cells.

Previous studies have demonstrated that the growth of tumor cells may be inhibited by antisense oligonucleotides (ASODNs) targeted against human telomerase (hTR) or human telomerase reverse transcriptase (hTERT), resulting in antitumor activity in a wide variety of tumors. However, few studies have investigated the effect of hTERT gene-targeted ASODNs on telomerase activity and cell proliferation in human esophageal cancer. In the present study, an MTT assay was used to determine the growth inhibition rate of Eca-109 cells treated with a hTERT-targeted phosphorothioate-ASODN (PS-ASODN). An inverted microscope was used to observe the morphologic changes of the cells following treatment with 5 µM PS-ASODN for 10 days. Telomerase activity was detected using the silver staining semi-quantitative telomeric repeat amplification protocol (TRAP) assay. Following treatment with the PS-ASODN (1-5 µmol/l), the proliferation of the Eca-109 cells was inhibited. The differences in inhibition rate between the PS-ASODN and blank control groups were statistically significant (P<0.05) when the concentration of the PS-ASODN was ≥2 µmol/l, whereas no statistically significant difference was identified between the non-specific-ASODN and blank control groups. The inhibition rate increased gradually as the concentration of the PS-ASODN increased and with time, suggesting that the PS-ASODN inhibited the growth of Eca-109 cells in a concentration-dependent, time-dependent and sequence-specific manner. The growth rate of the cells incubated with the PS-ASODN was reduced compared with that of the control cells. Cells treated with the PS-ASODN became round, suspended and reduced in size. The PS-ASODN was also found to inhibit telomerase activity. The ability of the PS-ASODN to inhibit the telomerase activity and cell proliferation of the Eca-109 cell line suggests that ASODNs have the potential to be novel therapeutic agents for the treatment of esophageal cancer.

Anticancer effect of salidroside on A549 lung cancer cells through inhibition of oxidative stress and phospho-p38 expression.

Oxidative stress is important in carcinogenesis and metastasis. Salidroside, a phenylpropanoid glycoside isolated from Rhodiola rosea L., shows potent antioxidant properties. The aim of the present study was to investigate the roles of salidroside in cell proliferation, the cell cycle, apoptosis, invasion and epithelial-mesenchymal transition (EMT) in A549 cells. The human alveolar adenocarcinoma cell line, A549, was incubated with various concentrations of salidroside (0, 1, 5, 10 and 20 µg/ml) and cell proliferation was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Propidium iodide (PI) staining was used to determine the cell cycle by flow cytometry. Cell apoptosis was detected by Annexin V-fluorescein isothiocyanate and PI double-staining, and tumor invasion was detected by Boyden chamber invasion assay. Western blot analysis was performed to detect the expression of EMT markers, Snail and phospho-p38. The results showed that salidroside significantly reduced the proliferation of A549 cells, inhibited cell cycle arrest in the G0/G1 phase and induced apoptosis. Salidroside inhibited transforming growth factor-ß-induced tumor invasion and suppressed the protein expression of Snail. As an antioxidant, salidroside inhibited the intracellular reactive oxygen species (ROS) formation in a dose-dependent manner in A549 cells, and depletion of intracellular ROS by vitamin C suppressed apoptosis by salidroside treatment. Salidroside was also found to inhibit the expression of phospho-p38 in A549 cells. In conclusion, salidroside inhibits cell proliferation, the cell cycle and metastasis and induces apoptosis, which may be due to its interference in the intracellular ROS generation, thereby, downregulating the ROS-phospho-p38 signaling pathway.

[Inheritance of the white-flower and its influence to related characters in eggplant (Solanum melongem Linn)].

The inheritance of the white-flower and its influence to related characters in eggplant were studied with the white-flowered mutant of XIANLVQIE and its maternal bred. Result showed that the flower color was attributed to a couple of complete dominance genes. Purple color flower "Col" was dominant to the white "col". Compared with the purple-flowered strain, the white-flowered strain not only grew more blooming, but also had more stamens in one flower, less pollens in one anther and less seeds in one fruit. In the meanwhile, in white-flowered eggplants, microspore and fruit was bigger, and yield was higher than purple-flowered. The white-flowered strain could be used as a variety and the white-flowered character could be used as a marker to appraise purity of hybrid in eggplant.

[In vitro killing effects of STI571 on esophageal carcinoma cell lines CE-48T and CE-81T].

BACKGROUND & OBJECTIVE: Tyrosine kinase mediates cell proliferation and differentiation, and plays important roles in tumorigenesis and development of esophageal carcinoma. STI571 is a tyrosine kinase inhibitor of platelet-derived growth factor receptor beta (PDGFR-beta) which is overexpressed in esophageal carcinoma. This study was to explore the in vitro killing effects of STI571 on esophageal carcinoma cell lines CE-48T and CE-81T. METHODS: The expression of PDGFR-alpha and PDGFR-beta in CE-48T and CE-81T cells was detected by Western blot. The killing effects of STI571 on CE-48T and CE-81T cells were evaluated by MTT assay. Cell apoptosis was analyzed by flow cytometry with Annexin V/PI labeling. The expression of p-PDGFR-beta was detected by Western blot before and after treatment of STI571. RESULTS: CE-48T cells expressed PDGFR-beta, but did not express PDGFR-alpha; CE-81T cells did not express both PDGFR-alpha and PDGFR-beta. The 50% inhibitory concentration (IC50) of STI571 was significantly lower for CE-48T cells than for CE-81T cells [(8.32+/-1.50) micromol/L vs. (41.02+/-7.64) micromol/L, P=0.002]. When treated with 10 micromol/L STI571 for 12 h, the apoptosis rate of CE-48T cells was (52.43+/-5.30)%, but the apoptosis rate did not increase as the treatment time and concentration increased. After treatment of STI571, the expression of p-PDGFR-beta was inhibited in CE-48T cells, but didn't change in CE-81T cells. CONCLUSIONS: STI-571 could induce the apoptosis of PDGFR-beta-positive esophageal carcinoma CE-48T cells. p-PDGFR-beta might be the target of STI571.

Negative implication of C-MYC as an amplification target in esophageal cancer.

Chromosomal aberrations (amplifications and deletions) underlie the genesis or development of cancer. Amplification of 8q24 is one of the most frequent events in esophageal cancer. To define whether C-MYC is the target gene for 8q24 amplification, we performed fluorescence in situ hybridization using a MYC (8q24.12 approximately q24.13) probe in esophageal cancer from southern China. Furthermore, we detected the expression status of several genes including C-MYC, TRIB1 (alias C8FW), and FAM84B (alias NSE2) in the regions of 8q24 via reverse transcriptase-polymerase chain reaction or immunohistochemical analysis (or both). Distinct amplification of 8q24 was found in esophageal carcinomas. Only 4 of 46 cases showed obvious protein expression in part of the esophageal cancerous nest. In particular, increased protein expression of C-MYC was shown only in a small part of a cancerous nest in the four cases. Positive C-MYC staining was detected mainly in the cytoplasm of esophageal cancer cells. No expression of TRIB1 was detected in esophageal squamous cell carcinomas. Of 59 cases, 39 (66%) cases showed increased expression of FAM84B in esophageal carcinomas. The results suggest that C-MYC and TRIB1 may not be the amplification target of 8q24 in esophageal cancer. FAM84B might be involved in the genesis or development of esophageal cancer in southern China. Whether FAM84B is the amplification target of esophageal cancer awaits further investigation.

[Clinical significance of hMAM mRNA detection in bone marrow of breast carcinoma patient].

OBJECTIVE: To investigate the expression of the human mammoglobin (hMAM) mRNA in bone marrow and its clinical significance in the breast cancer patient. METHODS: Expression of hMAM mRNA was detected using nested reverse transcription polymerase chain reaction (RT-PCR) in the bone marrow aspiration sample from 75 breast cancer patients, 15 patients with benign breast lesions and 8 healthy volunteers as control. The possible correlation of hMAM mRNA expression with clinico-pathological parameters and related molecular markers such as Ki67, p53 and VEGF were analyzed. RESULTS: The sensitivity of RT-PCR in this series reached 10(-6). The hMAM mRNA was found to be positively expressed by RT-PCR in 21 of 75 breast cancer patients with a positive rate of 28.0%. However, hMAM mRNA expression was not detected in the bone marrow aspiration samples from patients with benign breast lesions and healthy volunteers. The hMAM mRNA expression was positively correlated with axillary nodal involvement and progesterone receptor (PR) status (P < 0.05) as well as Ki67 expression in breast cancer tissue (chi2 = 4.936, P = 0.026), but not with age, tumor size, clinical stage, or estrogen receptor (ER) status (P > 0.05). CONCLUSION: RT-PCR is quite sensitive and has a high specificity in detecting the presence of hMAM mRNA in the bone marrow from breast cancer patients. Thereupon, hMAM mRNA may be useful as a molecular biomarker in detecting disseminated tumor cells (DTC) in the bone marrow of breast cancer patients. Positive hMAM mRNA expression result may have an impact upon therapeutic recommendations and patients' prognostic judgement.

[Methods of preserving intercostobrachial nerve during breast cancer operation and its clinical value].

OBJECTIVE: To study the methods and the clinical value of preserving intercostobrachial nerve during the axillary lymph nodes excision in breast cancer operations. METHODS: One hundred and sixty-two cases of stages I, II, IIIa breast cancer patients were divided into experimental group and control group respectively. The intercostobrachial nerves were preserved in experimental group and not in control group. Both groups were treated following the practice guideline of breast cancer, and found no recurrence during 4 to 36 months following up. RESULTS: The postoperative arm sensory disturbance was 22.2% in the experimental group, which was significantly different from that of the control group 73.3% (chi(2) = 41.80, P < 0.01), the incidence of pain is 12.5% in experimental group, which was also significantly different from that of control group 31.1% (chi(2) = 7.86, P < 0.01). CONCLUSION: Preserving intercostobrachial nerves may significantly decrease the postoperative morbidity of arm sensory disturbance and pain during axillary excision of stage I, II, IIIa breast cancer patients.

[Perioperative changes of cellular immune response of patients with esophageal carcinoma evaluated through detecting AgNOR content in peripheral blood T lymphocytes and T cell subsets].

BACKGROUND & OBJECTIVE: Patients with esophageal carcinoma often have immunosuppression. This study was to detect perioperative argyrophil nucleolar orgnizer region (AgNOR) content in peripheral blood T lymphocytes and T cell subsets of patients with esophageal carcinoma, investigate the influences of cellular immune condition and surgery on cellular immune function of these patients. METHODS: Peripheral blood samples were obtained from 75 healthy adults and 70 patients with esophageal carcinoma before surgery, 3 and 9 days after surgery. AgNOR content in peripheral blood T lymphocytes was measured by tumor immune microphotometry; T cell subsets was measured by flow cytometry. RESULTS: Before operation, AgNOR content, proportions of CD3(+)CD8(+) and CD8(+)CD25(+) T cells were significantly lower in patients than in healthy controls (P < 0.001); proportions of CD3(+) and CD3(+)CD4(+) T cells of patients were not significantly different from that of healthy controls (P > 0.05); proportions of CD4(+)CD25(+) and CD4(+)/CD8(+) T cells were significantly higher in patients than in healthy controls (P < 0.05). In peripheral blood T lymphocytes of the patients after operation, AgNOR content, CD3(+), CD3(+)CD4+(+), and CD4(+)/CD8(+) T cells were the lowest at the 3rd day, and the highest at the 9th day; CD3(+)CD8(+), CD4(+)CD25(+), and CD8+CD25+ T cells gradually ascended from the 3rd day. At the 9th day after operation, CD3(+) and CD3(+)CD4(+) T cells of patients were not significantly different from that of healthy controls (P> 0.05); AgNOR content, CD3(+)CD8(+) and CD8(+)CD25(+) T cells were still lower in patients than in healthy controls (P<0.05); CD4(+)CD25(+) and CD4(+)/CD8(+) T cells were still higher in patients than in healthy controls (P<0.001). CONCLUSION: Surgery injure may cause temporary cellular immunosuppression in patients with esophageal carcinoma, which slowly recovers early after operation.

[Vascular endothelial growth factor expression and microvessel density in stage I-II non-small cell lung cancer and their prognostic significances].

BACKGROUND & OBJECTIVE: TNM staging system is used widely to predict prognosis of non-small cell lung carcinoma (NSCLC) patients, but patients with the same stage may have very different survivals; better prognostic index is needed. Angiogenesis is considered to be essential for tumor development, progression, and metastasis, but the prognostic impacts of vascular endothelial growth factor (VEGF) and microvessel density (MVD) in NSCLC is controversial. This study was to evaluate the prognostic value of VEGF and MVD in NSCLC. METHODS: VEGF and MVD in 214 specimens of stageI-II NSCLC (20 in stage IA, 137 in stage IB, and 57 in stage IIB) were detected by tissue chip and SP immunohistochemistry. No patient underwent postoperative antitumor treatment. RESULTS: VEGF expression didn't relate to gender, age, blood type, pathologic type, and TNM stage (P0.05). MVD correlated with age and pathologic type (P0.05), but did not relate to gender, blood type, and TNM stage (P0.05). The mean value of MVD was 65.8+/-5.2 in VEGF-low patients, and 67.5+/-2.5 in VEGF-high patients (P0.05). The 5-year survival rate was significantly lower in MVD-high patients than in MVD-low patients (34.5% vs. 60.0%, P=0.013). Furthermore, multivariate Cox regression analysis showed that MVD (P=0.000) was an independent prognostic factor of NSCLC. CONCLUSION: High MVD closely relates to poor prognosis of stageI-II NSCLC.

[Expression and clinical significance of cytokeratin 19 in bone marrow of patients with breast cancer].

BACKGROUND & OBJECTIVE: Breast cancer may undergo metastasis in early phase. Distant metastasis, especially bone metastasis, may influence prognosis of breast cancer patients. Bone marrow micrometastasis (BMM) is difficult to detect with routine methods. This study was designed to evaluate expression and clinical significance of cytokeratin 19 (CK19) in bone marrow of patients with breast cancer. METHODS: Expression of CK19 mRNA in bone marrows of 65 breast cancer patients, 15 benign breast disease patients, and 8 healthy volunteers was detected by reverse transcription-polymerase chain reaction (RT-PCR). Correlation of CK19 mRNA expression to clinicopathologic features of the 65 breast cancer patients was analyzed. RESULTS: Positive rate of CK19 mRNA was 33.8% in the 65 breast cancer patients, and 0 in both benign breast disease patients and healthy volunteers. Expression of CK19 mRNA was positively correlated with tumor size and clinical stage (P < 0.05), but was not related to age and lymph node status (P > 0.05). In addition, positive rate of CK19 mRNA was positively correlated with carcinoembryonic antigen (CEA) in peripheral blood (r=0.372, P=0.002). CONCLUSIONS: CK19 mRNA may be used as a molecular marker to detect bone marrow micrometastasis in patients with breast cancer. The detection may help to select proper therapy and predict prognosis of breast cancer patients.

[Predictive value of detection of sentinel lymph nodes on lymphatic metastasis of non-small cell lung cancer].

BACKGROUND & OBJECTIVE: The metastasis status of regional lymph node is an important prognostic factor of non-small cell lung cancer (NSCLC). Sentinel lymph node (SLN) mapping and biopsy is a quick and high efficient technique to intraoperatively detect occult micrometastatic disease, however, its application in NSCLC is immature. This study was designed to investigate the feasibility of detecting SLN in patients with NSCLC during radical surgery, and to evaluate its accuracy of predicting metastasis status of regional lymph node. METHODS: Fifty patients with NSCLC underwent SLN detection. During radical operation, 4 ml of 1% isosulfan blue was injected into the lung tissue around the tumor at 3, 6, 9, and 12 o'clock sites. Location and number of blue dyed SLNs were recorded, and compared with pathologic results to calculate the accuracy and false negative rate of SLN detection. RESULTS: Blue dyed SLNs were seen in 33 patients with a detection rate of 66.0%. SLNs located in N1 lymph node of 24 patients (72.7%), in N2 lymph node of 6 patients (18.2%), in both N1 and N2 lymph nodes of 3 patients (9.1%). Approved by pathology, the accuracy of SLN detection was 87.9% (29/33), the sensibility was 73.3% (11/15), the false negative rate was 26.7% (4/15). CONCLUSION: SLN detection is valuable for predicting hilar and mediastinal lymph nodes metastases in NSCLC.

[Expression correlation of Ki67 to P53,VEGF,and C-erbB-2 genes in breast cancer and their clinical significances].

BACKGROUND & OBJECTIVE: The proliferating antigen Ki67 is an important biological marker in cell proliferation. This study was designed to investigate the expressions of Ki67, P53, vascular endothelial growth factor (VEGF),and C-erbB-2 in breast cancer tissue, and their correlations with cliniopathological significance. METHODS: The expressions of Ki67,p53,VEGF,and C-erbB-2 in 151 cases of breast cancer were assessed by immunohisto-chemistry, and their correlations with clinicopathological factors were statistically analyzed. RESULTS: The positive rate of Ki67 was 78.2%(113/151). The expression of Ki67 correlated with tumor stages (P< 0.05), but didn't relate to age, tumor size, and lymph node status (P >0.05). In addition,the expression of Ki76 had a positive correlation with the expressions of p53, and C-erbB-2 (P< 0.05), but not with the expression of VEGF (P >0.05). The co-expression of Ki67 and VEGF related to tumor size,and clinical stage (P< 0.05). CONCLUSIONS: The expression of Ki67 is an objective biological marker for estimating the occurrence and progression of breast cancer. Combined detection of Ki67 and VEGF may helps to judge the clinical stage of breast cancer.