Carnosol has been proved to have anti-breast cancer effect in previous research. But its ER subtype's specific regulation and mediation mechanisms remain unclear. The aim of this study is to observe the effect of carnosol on cell proliferation and its estrogen receptor α and β's specific regulation and mediation mechanisms with ER positive breast cancer T47D cell. With estrogen receptor α and β antagonists MPP and PHTPP as tools, the MTT cell proliferation assay was performed to observe the effect of carnosol on T47D cell proliferation. The changes in the T47D cell proliferation cycle were detected by flow cytometry. The effect of carnosol on ERα and ERβ expressions of T47D cells was measured by Western blot. The findings showed that 1 x 10(-5)-1 x 10(-7) mol x L(-1) carnosol could significantly inhibit the T47D cell proliferation, which could be enhanced by MPP or weakened by PHTPP. Meanwhile, 1 x 10(-5) mol x L(-1) or 1 x 10(-6) mol x L(-1) carnosol could significantly increase ERα and ERβ expressions of T47D cells, and remarkably increase ERα/ERβ ratio. The results showed that carnosol showed the inhibitory effect on the proliferation of ER positive breast cancer cells through target cell ER, especially ERβ pathway. In the meantime, carnosol could regulate expressions and proportions of target cell ER subtype ERα and ERβ.
Blotting, Western ; Breast Neoplasms ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Diterpenes, Abietane ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Estrogen Receptor Modulators ; pharmacology ; Estrogen Receptor alpha ; antagonists & inhibitors ; metabolism ; Estrogen Receptor beta ; antagonists & inhibitors ; metabolism ; Female ; Flow Cytometry ; Humans ; Molecular Structure ; Pyrazoles ; pharmacology ; Pyrimidines ; pharmacology

In order to understand the alcohol's toxicity to the quantitative alternations of synapses in mouse visual cortex, the expression of synaptophysin after prenatal alcohol exposure was investigated. In present study, the experimental mice at P0, P7, P14 and P30 were grouped, as control, 2 g x kg(-1) alcohol treatment and 4 g x kg(-1) alcohol treatment. The pre-synaptic elements which were used to represent synapses were marked with synaptophysin (a synaptic vesicle associated protein) by immunocytochemistry technique. The synaptophysin positive boutons in layer VI of visual cortex were imaged under laser confocal microscope. With stereological methods, the number cal density of synapse in visual cortex was calculated in different groups at various ages. Moreover, Western blotting was carried out to detect the expression of synaptophysin in visual cortex. The results showed that prenatal alcohol exposure could cause synaptic loss with long-term effect and in a dose dependent manner. For instance, there were significant difference among the different treatment groups of P0, P14 and P30 as well (P < 0.05). Western blotting supported the results of immunofluorescent labeling. In conclusion, prenatal alcohol exposure can induce the synaptic loss dose dependently and with long-term effect. Our findings implicate that the synaptic loss with long-term effect in CNS probably contributes to the lifelong mental retardation and memorial lowliness associated with childhood FAS.
Animals ; Dose-Response Relationship, Drug ; Ethanol ; administration & dosage ; toxicity ; Female ; Male ; Mice ; Mice, Inbred C57BL ; Microscopy, Confocal ; Pregnancy ; Prenatal Exposure Delayed Effects ; physiopathology ; Random Allocation ; Synapses ; drug effects ; Synaptophysin ; metabolism ; Visual Cortex ; drug effects ; physiopathology

OBJECTIVETo explore the method for obtaining olfactory ensheathing cells from human fetal olfactory mucosa by cell culture for selective adhesion in the presence of neurotrophin-3 (NT3) and low-concentration serum.

METHODSThe olfactory ensheathing cells were cultured alternatively in DMEM/F12 culture medium containing 10% fetal bovine serum (FBS) and the medium containing NT3 and 2.5% FBS every 72 h. The cells were observed for morphological changes and identified using immunocytochemistry with P75NTR and GFAP, and the cell purity was estimated.

RESULTSThe olfactory ensheathing cells from human fetal olfactory mucosa were positive for P75(NTR) and GFAP, and in in vitro culture, the cells exhibited dipolar or tripolar appearance with long thin neurites. On the 9th day of cell culture, the purity of the olfactory ensheathing cells reached about 83%.

CONCLUSIONThe olfactory ensheathing cells can be obtained by in vitro culture for selective adhesion in the presence of NT3 and low-concentration serum.

Cell Culture Techniques ; methods ; Cell Separation ; methods ; Cells, Cultured ; Culture Media ; Fetus ; Humans ; Neurotrophin 3 ; pharmacology ; Olfactory Bulb ; cytology ; Olfactory Mucosa ; cytology

Objective To evaluate the safety,efficacy and advantages of laparoscopic colorectal surgery for elderly patients with colorectal cancer.Methods Operation safety,complications,and postoperative recovery of 35 patients aged 70 years and over who underwent laparoscopic colorectal surgery(LAPA group) were analyzed retrospectively and compared with those of 78 elderly patients performed with conventional open surgery(OPEN group) and 35 patients younger than 65 years performed with laparoscopic colorectal surgery(LAPB group) between December 2002 and December 2003.Results No surgery-related death occurred in LAPA group and LAPB group,but 2 deaths were found in OPEN group because of severe pulmonary infection and anastomotic leakage respectively.There were 33 (42.3%) cases of complications in the OPEN group which were significantly more than those in LAPA and LAPB group (P0.05).Local recurrence rate,metachronous metastases rate and cumulative survival probability at 30 months were similar in the LAPA and OPEN groups.Conclusions Laparoscopic colorectal surgery is safe and beneficial to the patients aged 70 years and over and it could be adopted widely.

Objective To evaluate the laparoscopic management in diagnosing and treating small intestinal stromal tumors.MethodsSeventeen patients with small intestinal stromal tumors,who underwent laparoscopic diagnosis and surgical procedures,were retrospectively analyzed with the size of tumor,operative time,blood loss,length of incision,time for passage of flatus,post-operative hospital stay,operative complications and result of follow-up.Results All the 17 patients were diagnosed and treated by laparoscopic partial intestinal resections.The mean diameter of tumors was(3.6?1.3)cm,operative time(62.1?25.7)min,blood loss(17.1?15.2)mL,length of incision(3.4?1.1)cm,time for passage of flatus(2.2?1.2)d and post-operative hospital stay(7.8?2.0)d.As for the complications,one(5.9%)anastomosis bleeding was found and was treated by non-surgical procedure.After follow-up for 2-34 months,no recurrent tumor was found.Conclusion Laparoscopic exploration is useful to diagnose small intestinal stromal tumor accurately,and laparoscopic partial intestinal resection is a safe,effective and less invasive procedure for small intestinal stromal tumors.

Objective To evaluate the clinical application of laparoscopic local resection for gastric tumors.Methods Twenty-three patients with gastric tumors who were performed laparoscopic gastric local resection were retrospectively analyzed with the size of tumor,location of tumor,operative time,blood loss during the operation,time for passage of flatus,post-operative hospital stay,operative complications,post-operative pathological findings and result of follow-up.Results Twenty-three patients were successfully performed laparoscopic local resection,including 8 laparoscopic wedge resection(LWR)and 15 intragastric mucosal resections(IGMR),with no conversion to open surgery.The mean size of gastric tumor was(2.8?1.3)cm,the mean operative time was(82.2?35.5)min,the mean blood loss was(26.5?15.3)mL,the length of incision was(3.1?1.1)cm,the time for passage of flatus was(2.1?0.9)d,and the mean post-operative hospital stay was(7.8?2.0)d.Two patients(8.7%)were found with postoperative gastric mucosal blee-ding and were recovered well through non-operative treatment.The median time of follow-up was 12 months(2-45 months),and no recurrent tumor was observed.Conclusion Laparoscopic local resection is a feasible,safe,effective and less invasive procedure for gastric tumors.

OBJECTIVETo study the changes of lower esophageal sphincter (LES) high-pressure zone, and to determine the accurate length of myotomy on the esophageal and gastric sides.

METHODSThere were 15 patients undergoing the Heller's cardia-myotomies and Toupet fundoplications from May 2006 to December 2007. Among them, 9 patients were female and 6 was male. The age ranged from 28 to 61 years old, and the disease duration ranged from 6 months to 9 years. The intraoperative oesophageal manometry underwent in the surgical procedures to investigate the changes of the lower esophageal sphincter pressure and the length of myotomy.

RESULTSThere was no postoperative death. After (5.3 +/- 1.5) cm of esophageal side myotomy and (0.8 +/- 0.4) cm of gastric side myotomy, the mean LES pressure decreased from (33.6 +/- 13.3) mm Hg (1 mm Hg = 0.133 kPa) to (9.7 +/- 4.6) mm Hg and (4.8 +/- 3.1) mm Hg respectively (P < 0.05). The lower esophageal sphincter length ranged from 5 to 8 cm.

CONCLUSIONIntraoperative esophageal manometry helps determine the accurate myotomy length of myotomy on the esophageal and gastric sides of the gastroesophageal junction and provides valuable information for the Heller's myotomy.

Adult ; Esophageal Achalasia ; surgery ; Esophagus ; physiopathology ; surgery ; Female ; Humans ; Intraoperative Care ; Male ; Manometry ; Middle Aged ; Monitoring, Intraoperative

OBJECTIVETo investigate JAK2 exon 12 mutations in patients with Philadelphia (Ph) chromosome-negative myeloproliferative neoplasms (MPN) and the clinical characteristics of patients with JAK2 exon 12 mutants.

METHODSAllele-specific PCR (AS-PCR) was applied to identify JAK2 V617F mutation. Genomic DNA corresponding to exon 12 of JAK2 gene and epigenetic regulator gene (TET2, ASXL1, EZH2) were amplified by polymerase chain reaction (PCR). Identification of mutants was by direct sequencing and classification of mutation types by sequencing followed by plasmid cloning. SNP genotyping of two 46/1 tag SNPs, rs12340895 and rs10974944, was analyzed using commercially available Taqman assays on the 7500HT real-time PCR instrument according to standard protocols.

RESULTSNo JAK2 exon 12 mutation was detected in patients with ET, PMF or JAK2 V617F positive PV. Among 13 JAK2 V617F negative PV patients, JAK2 exon 12 mutation was detected as N542-E543del in 2(15.4%) patients who presented with a phenotype of predominant erythrocytosis and erythroid colonic grown from their bone marrow samples in the absence of exogenous EPO, reduced serum erythropoietin (EPO) level, and no mutations in TET2, ASXL1 or EZH2 genes. One of the affected patients was heterozygous for 46/1 but the second was negative for this haplotype.

CONCLUSIONThere was no need to detect JAK2 exon 12 mutation in ET, PMF or MPN-U patients without JAK2 V67F mutation. Ph negative MPN patients with JAK2 exon 12 mutations had somewhat unique clinical and laboratory features.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Marrow Neoplasms ; genetics ; DNA Mutational Analysis ; Exons ; Female ; Genotype ; Humans ; Janus Kinase 2 ; genetics ; Male ; Middle Aged ; Myeloproliferative Disorders ; genetics ; Philadelphia Chromosome ; Young Adult

OBJECTIVETo evaluate the clinical use of laparoscopy in diagnosis and surgical treatment in small intestinal tumors.

METHODSClinical data of 42 patients with small intestinal tumor undergoing laparoscopic diagnosis and surgical procedure from Sep. 2003 to Dec. 2005 were analyzed retrospectively. The operative time, blood loss,length of incision, time for passage of flatus, post-operative hospital stay and operative complications were evaluated.

RESULTSAll the patients were diagnosed and treated by laparoscopic procedure successfully, including 4 laparoscopic local resection of the tumors, 36 laparoscopy-assisted partial intestinal resections, 1 right hemicolectomy, and 1 laparoscopic exploration. The mean operative time was (73.1+/-32.9) min, the mean blood loss was (20.7+/-31.2) ml, the mean length of incision was (3.7+/-1.2) cm, the mean time for passage of flatus was (2.2+/-0.8) d, and the post-operative hospital stay was (8.0+/-3.1) d. Postoperative complications occurred in 2 patients (4.8% ) including anastomosis bleeding and adhesive intestinal obstruction in one case respectively. After follow-up from 3 to 30 months, no recurrent tumor was found except one patient with advanced intestinal carcinoma.

CONCLUSIONLaparoscopic exploration can diagnose small intestinal tumors accurately, and laparoscopy-assisted surgical treatment is safe with less trauma and fast recovery.

Adult ; Aged ; Female ; Humans ; Intestinal Neoplasms ; diagnosis ; surgery ; Intestine, Small ; Laparoscopy ; Middle Aged ; Retrospective Studies

OBJECTIVETo evaluate laparoscopic surgery for gastric and small intestinal stromal tumors.

METHODSThe clinical data of patients with laparoscopic resection of gastric and small intestinal tumors, admitted to our center from Dec. 2003 to Jul. 2006, were retrospectively analyzed. The data included the surgical procedure, operative time, blood loss, length of incision, time for passage of flatus, postoperative length of stay, operative complications, pathology and the results of follow-up.

RESULTSAll the 33 patients with gastric and small intestinal stromal tumors were treated by laparoscopic procedures successfully, including 8 laparoscopic gastric wedge resections, 8 laparoscopic transgastric tumor-everting resections, and 17 laparoscopic segmental resections of small intestine. The mean operative time was (73.1+/-27.0) min, the mean blood loss was (19.8+/-14.0) ml, the mean length of incision was (3.3+/-1.1) cm, the mean time for passage of flatus was (2.2+/-1.0) d and the postoperative length of stay was (8.1+/-2.0) d. The bleeding of intestinal anastomosis and gastric mucosa occurred on 3 patients (9.1%), who were healed with conservative therapy. No recurrence was found during 2-33 months follow-up.

CONCLUSIONLaparoscopic surgery is an effective, safe, less invasive procedure for treatment of gastric and small intestinal stromal tumors.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; surgery ; Humans ; Intestine, Small ; Laparoscopy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Young Adult