OBJECTIVETo isolate breast cancer stem cells from breast cancer patients and identify their biological characteristics.

METHODSMammospheric cells were purified and enriched from the tumor tissues of breast cancer patients using mammosphere culture. Their expressions of CD44 and CD24 were analyzed by flow cytometry, and ALDH1, ESA and Oct4 expressions were determined by Western Blotting. The primary mammospheric and adherent cells, at the density of 2×10(4), 2×10(5) or 2×10(6), were inoculated into NOD/SCID mice to observe their tumorigenic and metastatic activities.

RESULTSWith mammosphere culture method, 62.36% of the mammospheric cells showed CD44(+)/CD24(-/low) phenotype. The expressions of ALDH1, ESA and Oct4 in the mammospheric cells were significantly higher than those in the adherent culture-derived breast cancer cells (P<0.05). Primary mammospheric cells were at least 100-fold more tumorigenic than the adherent cells; the mammospheric cells were associated with liver or lung metastases, but the adherent cells were not.

CONCLUSIONMammosphere culture can be employed to obtain breast cancer stem cells from the tumor tissues of breast cancer patients.

Adult ; Animals ; Aryldialkylphosphatase ; metabolism ; Breast Neoplasms ; pathology ; CD24 Antigen ; metabolism ; Cell Culture Techniques ; methods ; Female ; Humans ; Hyaluronan Receptors ; metabolism ; Isoenzymes ; metabolism ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Middle Aged ; Neoplastic Stem Cells ; cytology ; metabolism ; Octamer Transcription Factor-3 ; metabolism ; Primary Cell Culture ; Retinal Dehydrogenase ; metabolism

ObjectiveTo evaluate the value of intraoperative frozen section examination (IFS) in the diagnosis and surgical procedures selection for renal occupying lesions. MethodsFrom January 2006 to December 2010,IFS was used in 114 men and 81 women with renal occupying lesions.The mean age was 52 years (range 17 -78).In 104,89,and 2 patients,lesions were in the right,left and bilateral kidneys,respectively.All patients underwent physical examination,129 were asymptomatic at presentation while clinical symptoms were observed in 66.The largest dimension of the tumors were 4 cm or less in 128 patients,4- 7 cm in 49,and larger than 7 cm in 18,respectively.The outcomes between IFS and postoperative routine paraffin section examination were compared.In cases with renal tumors nephrectomy or partial nephrectomy was performed.The results of IFS were compared between the 2 groups. ResultsThe sensitivity and specificity of IFS for renal malignant lesions was 96.6％ and 100％,respectively.The total accuracy rate of IFS for renal occupying lesions was 97.4％.By subgroup analysis,the accuracy rate of clear cell carcinoma,papillary cell carcinoma,chromophobe cell carcinoma,sarcomatoid cancer,nephroblastoma,benign tumor and urothelial cancer was 94.3％,25.0％,16.7％,0,0,97.6％ and 100.0％,respectively.Partial nephrectomy and nephrectomy were performed in 57 and 123 patients with renal tumors,respectively.The surgical procedures selection was significantly associated with the lesion size (4 cm or less for 80.7％ vs 62.6％,P =0.015) and the malignant lesion diagnosed by IFS (31.6％ vs 93.5％,P＜0.001). Conclusion The accuracy of frozen section analysis for renal malignant lesions during surgery is reliable and significantly high,and the results can exert an important impact on surgical procedures selection.

BACKGROUNDOverexpression of G-protein coupled receptor 34 (GPR34) affects the progression and prognosis of human gastric adenocarcinoma, however, the role of GPR34 in gastric cancer development and progression has not been well-determined. The current study aimed to investigate the effect of GPR34 knockdown on the proliferation, migration, and apoptosis of HGC-27 gastric cancer cells and the underlying mechanisms.

METHODSThe expression of GPR34 in gastric cancer cell line HGC-27 was detected by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. HGC-27 cells were employed to construct the stable GPR34 knockdown cell model in this study. Real-time RT-PCR and Western blotting were applied to validate the effect of short hairpin RNA (ShRNA) on the expression of GPR34 in HGC-27 gastric cells. The proliferation, migration of these cells were examined by Cell Counting Kit-8 and transwell. We also measured expression profile of PI3K/PDK1/AKT and ERK using Western blotting.

RESULTSThe ShRNA directed against GPR34 effectively inhibited both endogenous mRNA and protein expression levels of GPR34, and significantly down-regulated the expression of PIK3CB (P < 0.01), PIK3CD (P < 0.01), PDK1 (P < 0.01), phosphorylation of PDK1 (P < 0.01), Akt (P < 0.01), and ERK (P < 0.01). Furthermore, GPR34 knockdown resulted in an obvious reduction in HGC-27 cancer cell proliferation and migration activity (P < 0.01).

CONCLUSIONSGPR34 knockdown impairs the proliferation and migration of HGC-27 gastric cancer cells in vitro and provides a potential implication for therapy of gastric cancer.

Apoptosis ; genetics ; physiology ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; genetics ; physiology ; Humans ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Receptors, Lysophospholipid ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism

OBJECTIVETo analyze the causes of the complications associated with the pedicle screw fixation during and after operation, as well as the methods for prevention and treatment.

METHODSFrom January 1988 to December 1999, 475 patients who underwent pedicle screw fixation for various spinal disorders were reviewed retrospectively with statistical analysis.

RESULTSFour hundred and twenty-eight patients (90.1% of all) were followed up for (6.0 +/- 3.7) years. Complications of CD, DRFS, Dick, RF and steffee among 72 cases (80% of all) occurred during the operation of pedical screw fixation in 26 cases with the incidence of 5.5% and after the operation in 64 cases with the incidence of 15%. The incidence of operative/postoperative complications were 0-12.9% in CD, 6.8%-11.9% in DRFS, 7.3%-12.7% in Dick, 2.4%-19.5% in RF, and 10.1%-21.5% in Steffee respectively. The main complications included pseudoarthrosis, pedicle screw broken, pedicle screw malposition and so on. The complications were mainly due to unskilled technique of pedicle screw fixation, implant defect in design and so on.

CONCLUSIONSThe complications associated with the technique of pedicle screw fixation must not be neglected. The main causes are laid on unskilled technique of pedicle screw fixation and implant defect in design.

Equipment Failure ; statistics & numerical data ; Female ; Follow-Up Studies ; Humans ; Incidence ; Internal Fixators ; Male ; Postoperative Complications ; epidemiology ; etiology ; Pseudarthrosis ; epidemiology ; etiology ; Retrospective Studies ; Spinal Diseases ; surgery ; Spinal Fusion ; methods

OBJECTIVETo observe the therapeutic effect of acupuncture at different stages on pseudobulbar palsy. Methods Two hundred and forty cases of pseudobulbar palsy were divided into 4 groups according to different courses of disease, i.e. group I, the course within 10 days; group II, between 10-30 days; group III, between 1-3 months; group IV, between 3-6 months. They were treated with acupuncture at Fengchi (GB 20) for 2 courses, and then their therapeutic effects were ohserved.

RESULTSThe effective rate was 100.0% in the group I, 96.7% in the group II, 83.3% in the group III and 76.7% in the group IV, with a significant difference among the 4 groups (P < 0.01).

CONCLUSIONAcupuncture at Fengchi (GB 20) at any stage has therapeutic effect on pseudohulbar paisy, hut the earlier treatments, the better the therapeutic effects.

Acupuncture Points ; Acupuncture Therapy ; methods ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Pseudobulbar Palsy ; therapy

OBJECTIVETo investigate the expression of MAGE-B genes in hepatocellular carcinoma (HCC) in order to find new targets for immunotherapy.

METHODSThe expression of MAGE-B1, B2, A1 and A3 mRNA was detected using RT-PCR in HCC tissues and the corresponding adjacent non-HCC tissues from 47 HCC patients, 30 samples of cirrhosis and normal liver tissues. Four samples selected randomly from MAGE-B1 or B2 with positive RT-PCR results were sequenced to confirm the results of RT-PCR. The relationship between the expression of MAGE-B and some clinicopathological parameters was analyzed.

RESULTSMAGE-B1 mRNA and MAGE-B2 mRNA were detected in 44.7% (21/47) and 61.7% (29/47) of HCC samples, respectively, while neither MAGE-B1 nor MAGE-B2 could be detected in the corresponding adjacent non-HCC liver tissues. In addition, none of 30 samples of cirrhosis and normal liver tissues was shown to express both MAGE-B genes. The DNA sequence confirmed that the RT-PCR products were truly target cDNA. The frequency of the expression of MAGE-A1 and A3 was 74.5% (35/47) and 44.7% (21/47), respectively. There was significant correlation between the expression of MAGE-B and MAGE-A (P < 0.05). However, the positive expression of MAGE-B was observed in 5 out of 12 HCC tissues without expression of MAGE-A1 and/or A3. When all four MAGE genes were examined, the positive rate of expression of one, two, three and four genes was 83.0% (39/47), 55.3% (26/47), 48.9% (23/47), and 38.3% (18/47) of 47 HCC tissues, respectively. No correlation was found between the expression of MAGE-B and clinical parameters such as age, sex, tumor size, degree of tumor differentiation, serum alpha-fetoprotein level and hepatitis B virus or hepatitis C virus infection (P > 0.05).

CONCLUSIONMAGE-B genes are expressed with relatively high frequency and specificity in HCC. Most HCC patients with positive expression of at least one member of MAGE-B or MAGE-A gene family are adequate candidates to receive specific immunotherapy. Frequent co-expression of multiple members of MAGE-B and MAGE-A subfamilies provides the possibility of using polyvalent vaccines to achieve more effective immunotherapeutic results.

Antigens, Neoplasm ; genetics ; Carcinoma, Hepatocellular ; genetics ; Gene Expression ; Humans ; Liver Neoplasms ; genetics ; Melanoma-Specific Antigens ; Neoplasm Proteins ; genetics ; Prognosis ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the effect of inversion-table in the treatment of lower pole renal stones.

METHODSFrom March 2006 to September 2008, 36 patients with lower pole renal calculi were treated by inversion-table, 13 cases with stone diameter from 4 mm to 20 mm received extracorporeal shock wave lithotripsy (ESWL) combined with inversion-table treatment and other 23 cases with stone diameter from 2 mm to 4mm received inversion-table treatment alone.

RESULTSAll of 36 cases underwent inversion-table treatment successfully and no one had cardio-cerebral accident. After following up 3 months, the radiologically documented complete stone clearance rate at 3 months group was 83.3%.

CONCLUSIONInversion-table treatment is a safe and valuable adjunct in assisting passage of lower pole renal stone fragments after mini-invasive treatment of stone and can improve stone-free rate.

Adult ; Female ; Follow-Up Studies ; Humans ; Kidney Calculi ; therapy ; Lithotripsy ; Male ; Middle Aged ; Posture ; Treatment Outcome ; Vibration ; therapeutic use

OBJECTIVETo investigate the efficacy and side effect of topical beta-blocker (Timolol Maleate) in the treatment of periocular hemangioma in a prospective study.

METHODS432 outpatients with infantile hemangioma visited our special clinic service in Shanghai Ninth People's Hospital from July 2010 to December 2011. Among them, 12 superficial periocular lesions were selected in the study. Timolol was used topically on the lesion in every 12 hours. Two independent special doctors evaluated the results according to the pictures before and after four-week application of timolol.

RESULTSWere categorized into four levels: continuous growth (the lesion continues to grow), stable (no visible change), moderate (0-50% of regression) , perfect (more than 50% of improvement). Result of the 12 outpatients, 4 showed perfect result, 2 moderate, 4 stable and 2 continuous growth. No side effect was observed.

CONCLUSIONSTopical timolol is effective and safe in the treatment of superficial periocular infantile hemangioma. It could be considered as the first line treatment of proliferative superficial hemangioma.

Administration, Topical ; Angiogenesis Inhibitors ; therapeutic use ; China ; Facial Neoplasms ; drug therapy ; pathology ; Hemangioma ; drug therapy ; pathology ; Hemangioma, Capillary ; drug therapy ; pathology ; Humans ; Infant ; Prospective Studies ; Skin Neoplasms ; drug therapy ; pathology ; Timolol ; therapeutic use ; Treatment Outcome

OBJECTIVETo prospectively assess the efficacy and safety or propranolol as a first-line treatment for problematic infantile haemangioma in China.

METHODSFrom Mar. 2009 to Feb. 2010, 78 patients with problematic infantile hemangioma were included in the prospective study. The characteristics of the tumor, including sex, age, site, complications, were recorded. The response to treatment at 1 week, at 1 month and at the end of treatment was evaluated. The efficacy of treatment was graded as no response, stabilization, or accelerated regression. The indications for treatment, side effects and relapse after treatment were documented. The mean follow-up period was 16.7 months (range, 12.1-23.6 months).

RESULTSOral therapy was initiated at mean age of 3.7 months (range, 1.1-9.2 months) as first-line therapy. The mean age at the end of treatment was 11.2 months (range, 5.2-22.3 months). The treatment was lasted for 7.6 months (range, 2. 1-18.3 months). One week after treatment beginning, the hemangioma growth was controlled in all the patients. The accelerated regression was achieved in 88.5% (69/78) of patients after one week of treatment, and 98.7% (77/78) of patients after 1 month of treatment and at the end of treatment. Ulceration was occurred in 14 cases before treatment, which was healed after treatment for 2 months. Minor side effects were happened in 15.4% (12/78) of patients. Rebound growth of lesion was noticed in 35.9% (28/78) of patients.

CONCLUSIONSPropranolol is effective in the treatment of infantile hemangioma with minor side effect. We suggest it should be used as the first-line treatment.

Female ; Follow-Up Studies ; Hemangioma ; drug therapy ; Humans ; Infant ; Male ; Propranolol ; administration & dosage ; therapeutic use ; Prospective Studies ; Treatment Outcome

OBJECTIVETo analyze the gene expression level of fibroblast activation protein in HBV related hepatocellular carcinoma patients and discuss its clinical significance.

METHODSFAP gene expression in 33 hepatocellular carcinoma patients cancer tissues, peficancerous tissues, distant relative normal liver tissues and 13 normal liver tissues were examined by reverse transcription PCR; and real-time fluorescent quantitative PCR (qRT-PCR) was used to quantify their expression.

RESULTSFAP were expressed in all the tissues,the relative expression values in cancer tissues, peficancerous tissues and distant relative normal liver tissues were 5.14 +/- 6.69, 1.58 +/- 0.96, 1.63 +/- 0.94, respectively, the differences were statistically significant (F = 4.401, P < 0.05); and in TNM stage I, II, IIII, they were 2.89 +/- 3.35, 4.15 +/- 4.69, 10.09 +/- 9.51 respectively; in well-differentiated, differentiated and poorly differentiated hepatocellular carcinoma were 1.62 +/- 1.74, 3.84 +/- 3.79, 1.26 +/- 13.34 respectively. The differences were all statistically significant (P < 0.05).

CONCLUSIONFAP may play an important role in the occurrence and development of HBV related hepatocellular carcinoma.

Adult ; Aged ; Carcinoma, Hepatocellular ; etiology ; metabolism ; Female ; Gelatinases ; genetics ; physiology ; Gene Expression Regulation, Neoplastic ; Hepatitis B ; complications ; Humans ; Liver Neoplasms ; etiology ; metabolism ; Male ; Membrane Proteins ; genetics ; physiology ; Middle Aged ; RNA, Messenger ; analysis ; Serine Endopeptidases ; genetics ; physiology