PURPOSE: Laparoscopic cholecystectomy (LC) has been accepted as one of the best methods for treatment of patients with gallbladder (GB) disease. Diagnosis of malignant gallbladder disease by radiologic finding is possible; however, some cases can be diagnosed incidentally as cancer of the gallbladder after a LC. Prognosis of GB cancer is very poor; however, a complete recovery can be achieved with early detection and resection. This study was performed in order to determine incidence and clinical features of incidentally detected GB cancer after LC. METHODS: A retrospective analysis of 10 Patients diagnosed as GB polyp prior to surgery, but diagnosed incidentally as GB cancer after LC from January 2002 to February 2011 was conducted. RESULTS: Of 124 cases of GB polyp, 10 patients were diagnosed as incidental GB cancer, with incidence rate of approximately 8%. Mean age was 60.9. Mean diameter of polyps was 1.1 cm, with six cases of pedunculated polyps, and four cases of sessile polyps. GB stones were diagnosed by radiologic study prior to surgery in four patients. Four patients had pT1a tumors (limited to mucosa) and six patients had pT2 (invaded perimuscular tissue) or pT3 (perforated serosa, direct invasion into liver). CONCLUSION: Malignancy potential of GB polyps was 8%. Surgeons should be aware of the possibility of malignancy and frozen section biopsy should be performed for examination of suspected GB tissue. If needed, radical cholecystectomy should be performed.
Biopsy ; Cholecystectomy ; Cholecystectomy, Laparoscopic ; Frozen Sections ; Gallbladder ; Gallbladder Diseases ; Gallbladder Neoplasms ; Humans ; Incidence ; Polyps ; Prognosis ; Retrospective Studies ; Serous Membrane

OBJECTIVE: To evaluate the application of MSCT and post-processing images to fractures of nasal bone in forensic identification. METHODS: 134 cases were examined by thin slice scanning with MSCT and all of the data were sent to workstation for MPR and SSD. The result of MSCT was compared with that of X-ray. RESULTS: There are 55 (41.04%) cases of linear fracture, 46 (34.33%) cases of comminuted fracture, 27 (20.15%) cases of depressed fracture and 6 (4.48%) cases of no fracture in this sample. With X-ray or CR, 48 (35.82%) cases were misdiagnosed or underdiagnosed. 133 (99.25%) cases were confirmed by MSCT. Significance difference was found between X-ray and MSCT (chi2= 45.0816, P<0.001). CONCLUSION MSCT and post-processing images might be the chief evidence for nasal fractures in forensic identification.
Adolescent ; Adult ; Chromatography, Thin Layer ; Female ; Forensic Medicine/methods* ; Fractures, Comminuted/diagnostic imaging* ; Humans ; Imaging, Three-Dimensional ; Male ; Middle Aged ; Nasal Bone/injuries* ; Skull Fractures/diagnostic imaging* ; Tomography, Spiral Computed/methods* ; Young Adult

Hemodynamics changes in pregnant women,cardiac output increases by 50％,increasing the burden of cardiovascular system.It leads to further aggravation of original cardiovascular diseases during pregnancy,and non-pregnancy potential cardiovascular diseases will show up.Advanced age,obesity and diabetes etc.are risk factors of cardiovascular diseases,and they are more common in pregnant women than those non-pregnant,thus incidence rate of cardiovascular diseases increase in pregnant women consequently,making 1％～4％pregnancy more compli-cated than the normal pregnancy,which is also an important reason leading to pregnant woman death and adverse outcomes such as premature birth and stillbirth.Treatment during pregnancy should consider impact of fetus,there-fore its more complicated and difficult for clinical treatment of cardiovascular diseases during pregnancy.The pres-ent article mainly elaborates influence of cardiovascular diseases on pregnancy,including gestational hypertension,gestational diabetes and perinatal cardiomyopathy.

OBJECTIVETo identify the distribution and differentiation of ABL kinase domain mutation in the Chinese Han nationality imatinib resistant chronic myeloid leukemia (CML) and Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+)ALL).

METHODSBone marrow or peripheral blood samples of 112 imatinib resistant CML patients and 21 Ph(+)ALL patients were obtained from the first affiliated hospital of Soochow university according to local law. Total RNA was extracted from the mononuclear cells using a TRIzol reagent. ABL kinase domain (KD) mutation was detected by direct sequencing.

RESULTSOf the 112 imatinib resistant CML patients, 54.46%(61 cases) had ABL KD mutation. Twenty-three mutants were identified in 20 amino acid sites and 23.21% (26 cases) ABL KD mutations were in P-loop region. ABL KD mutations were also detected in 71.43% (15 cases) imatinib resistant Ph(+)ALL patients, with 10 mutations in 8 amino acid sites. The most frequent mutation was T315I (28.57%), followed by E255K/V (19.05%) and Y253F/H (14.29%). The frequency of T315I was much higher in imatinib resistant Ph(+) ALL than that in imatinib resistant CML (P = 0.001). Ph(+)ALL with additional chromosomal aberrations also had a higher rate of ABL KD mutation than that of CML (P = 0.010). Ph(+)ALL gained ABL KD mutation faster than CML (P < 0.010).

CONCLUSIONChinese imatinib resistant CML and Ph(+)ALL patients had different characteristics in ABL KD mutation. The rate of ABL KD mutation in Ph(+)ALL with additional chromosomal aberrations was much higher than that of CML with additional chromosomal aberrations.

Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Benzamides ; pharmacology ; Chromosome Aberrations ; Drug Resistance, Neoplasm ; genetics ; Female ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Middle Aged ; Mutation ; Philadelphia Chromosome ; Piperazines ; pharmacology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Protein-Tyrosine Kinases ; genetics ; Proto-Oncogene Proteins c-abl ; genetics ; Pyrimidines ; pharmacology ; Young Adult

OBJECTIVETo observe the effects of Stragalus membranaceus injection on nitric oxide and endothelin levels of intestinal mucosa in reperfusion injury after hemorrhage shock.

METHOD32 SD rats were randomly divided into four groups: normal group, model group, low dosage group, (treated with Astragalus membranaceus 10 g x kg(-1)); high dosage group (treated with Astragalus membranaceus 20 g x kg(-1)). Models of hemorrhagic shock for 60 minutes and reperfusion for 90 minutes were created. The animals were administrated 3 mL therapeutic solution before reperfusion. At the end of study, intestinal pathology was observed, and the concentration of lactic acid (LD), nitric oxide (NO), endothelin (ET) of intestinal mucosa were detected.

RESULTThe intestinal pathology showed that intestinal mucosa epithelial cells damage in model group was severe, in low dosage group was medium, in high dosage group was slight, and no obvious damage was found in normal group. The concentration of LD and NO of small intestine mucous membrane in model group and low dosage group were significantly higher than those in high dosage group and normal group (P < 0.05), but there were no significant differences between high dosage group and normal group (P > 0.05). The concentration of ET of small intestine mucous membrane in model group was the highest of the four groups (P < 0.05). The concentration of ET in low dosage group was significantly higher than that in high dosage group and normal group (P < 0.05), but there were no significant differences between high dosage group and normal group (P > 0.05).

CONCLUSIONStragalus membranaceus injection can reduce small intestine mucous damage by protecting endothelium function in injury after hemorrhage shock-reperfusion.

Animals ; Astragalus membranaceus ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Endothelins ; metabolism ; Ileum ; metabolism ; pathology ; Injections, Intravenous ; Intestinal Mucosa ; metabolism ; pathology ; Lactic Acid ; metabolism ; Male ; Nitric Oxide ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; metabolism ; pathology ; Shock, Hemorrhagic ; complications

OBJECTIVETo evaluate the prevalence of TET2 gene mutation in acute myeloid leukemia (AML) patients, and analyze their clinical characteristics and prognosis.

METHODSPolymerase chain reaction (PCR) and direct sequencing were used to sequence exon 3 to 11 of TET2 gene.

RESULTSAmong 96 AML patients, TET2 gene mutation was detected in 13 (13.54%) patients (95%CI 6.70% - 20.38%). The median age was 54 years in mutated group and 41 years in unmutated group (P = 0.010). Mutated and unmutated patients did not significantly differ in gender, white blood cells (WBC) count at diagnosis, platelet count, PB and BM blast percentage and chromosome karyotype, excepting for hemoglobin level 84 (70 - 108) g/L in mutated group versus 70 (55 - 87) g/L in unmutated group (P = 0.032). TET2 gene mutation had no significant correlation with C-KIT, FLT3, JAK2V617F mutations, but did with NPM1 mutation. TET2 mutated patients had lower CR1 rate and 2-year overall survival than unmutated in non-M(3) patients (P < 0.05).

CONCLUSIONSTET2 gene mutation is more prevalent in older AML patients and has a certain correlation with clinical characteristics and outcome. It may be a molecular marker for poor prognosis in AML.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA Mutational Analysis ; DNA-Binding Proteins ; genetics ; Exons ; Female ; Humans ; Karyotype ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Proto-Oncogene Proteins ; genetics ; Young Adult

OBJECTIVETo assess the prevalence of several tyrosine kinases (TKs) gene mutations including c-Kit, FLT3 and JAK2 V617F in core binding factor related acute myeloid leukemia (CBF-AML), and analyze their impact on clinical characteristics and prognosis.

METHODSMutations of c-Kit, FLT3-ITD and FLT3-TKD were detected by genomic DNA PCR and sequencing, and JAK2 V617F mutation screening by allele-specific PCR in 58 newly diagnosed CBF-AML patients [28 AML with inv(16) and 30 with t(8;21)], and analyze the patients clinical characteristics and prognoses.

RESULTSc-Kit aberrations were detected in 32.8% cases, including 6 cases mutated in exon 8 (mutKIT8) and 13 mutated in exon 17 (mutKIT17). MutKIT8 was more prominent in inv(16) than in t(8;21) patients (21.4% vs 0, P = 0.009). Only 2 cases had FLT3-ITD and 7 (12.1%) FLT3-TKD mutations. The result of JAK2 V617F mutation screenings in these CBF-AML patients was negative. The frequency of receptor tyrosine kinases(RTK) mutations was 46.6% and only one case had two kinds of missense mutations (mutKIT8 & TKD(+)). Median age of onset was higher for mutKIT17 than for wide-type c-Kit (wtKIT) patients (55 vs 31, P = 0.003). c-Kit mutations were significantly associated with decreased overall survival (OS) and continuous complete remission (CCR) rates (P = 0.053, and 0.048 respectively), and so did more for exon17 mutated patients reduced (P = 0.005, and 0.013 respectively). FLT3-TKD mutation showed no effects on prognosis of CBF-AML patients.

CONCLUSIONSRTK mutations are common in patients with CBF-AML. c-Kit mutations frequently and JAK2V617F mutation rarely appear in CBF-AML. c-Kit mutations, especially mutKIT17 confers higher relapse risk and poorer prognosis.

Adolescent ; Adult ; Aged ; Core Binding Factors ; DNA Mutational Analysis ; Female ; Humans ; Janus Kinase 2 ; genetics ; Leukemia, Myeloid, Acute ; diagnosis ; etiology ; genetics ; Male ; Middle Aged ; Mutation ; Prognosis ; Protein-Tyrosine Kinases ; genetics ; Proto-Oncogene Proteins c-kit ; genetics ; Young Adult ; fms-Like Tyrosine Kinase 3 ; genetics

OBJECTIVETo evaluate the prevalence and distribution of C-kit, NPM1 and FLT3 gene mutations in patients with acute myeloid leukemia (AML), and to analyze the relationship between the gene mutations and their prognosis.

METHODSMutations in exon 8 and 17 of C-kit gene, exon 12 of NPM1 gene, exon 20 of FLT3-TKD gene, and exon 14/15 of FLT3-ITD gene were detected by direct sequencing. Clinical data was collected and followed up if the patient had accepted treatment in our hospital.

RESULTSAmong the 656 AML patients, mutations in C-kit exon 8 were found in 6 patients (0.9%), C-kit exon 17 in 33 (5.0%), NPM1 in 169 (25.8%), FLT3-TKD in 46 (7.1%), and FLT3-ITD in 178 (27.1%). Six subtypes of mutations were detected in C-kit exon 8, 8 in C-kit exon 17, 11 in FLT3-TKD, 15 in NPM1, of which 5 were not reported before. C-kit exon 17 mutations were more frequently detected in patients with t(8;21) and exon 8 in patients with inv(16) cytogenetic abnormality. No other gene mutations except FLT3 were detected in M(3) patients. NPM1 and ITD mutations were often detected in individuals with normal cytogenetics or M(5) and M(1) of FAB classification, and accompanied with high white blood cell counts in peripheral blood, high blast counts in bone marrow and low CD34 expression. The older the patients were when diagnosed, the more gene mutations and the higher white blood cell count were detected. More mutations were found in individuals with normal karyotype than that with other karyotypes. It appeared that FLT3-ITD was significantly associated with shorter overall survival (OS) (P = 0.004), NPM1 was not significantly associated with OS, but NPM1(+)/ITD(-) patients had the longest OS.

CONCLUSIONSOur results showed that the mutation types and amounts had particular distribution in MICM subtypes, and were associated with white blood cell counts in peripheral blood, blast counts in bone marrow and prognosis. Especially for patients with normal karyotype, the genetic mutations could be new molecule marker.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; DNA Mutational Analysis ; Female ; Humans ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; Male ; Middle Aged ; Mutation ; Nuclear Proteins ; genetics ; Prognosis ; Proto-Oncogene Proteins c-kit ; genetics ; Young Adult ; fms-Like Tyrosine Kinase 3 ; genetics

BACKGROUNDCervical keratinocytes are recovered at a low numbers and frequently associated with contaminating human fibroblasts which rapidly overgrow the epithelial cells in culture with medium supplemented with 10% fetal bovine serum (FBS). However, it is difficult to initiate keratinocyte cultures with serum-free keratinocyte growth medium alone because cell attachment can be poor. Therefore, the culture of these cells is extremely difficult. In this study, we described a modified culture medium and coated culture plastics for growing normal human cervical epithelial cells in vitro.

METHODSNormal cervical epithelial tissue pieces were obtained and digested with type I collagenase to dissociate the cells and a single cell suspension produced. The cells were cultured on plastic tissue culture substrate alone or substrate coated with collagen type I from rat tail, with modified keratinocyte serum-free medium (K-SFM) supplemented with 5% FBS. After attachment, the medium were replaced with K-SFM without FBS. The expression of basal keratins of the ectocervical epithelium, K5, K14 and K19 were assayed by immunofluorescence with monoclonal antibodies to identify the cell purity.

RESULTSOur results indicate that cells attached to the culture plastic more quickly in K-SFM supplemented with 5% FBS than in K-SFM alone, as well as to tissue culture plastic coated with collagen type I than plastic alone. The modified medium composed of K-SFM and 5% FBS combined with a specific tissue culture plastic coated with collagen type I from rat tail was the best method for culture of normal cervical epithelial cells. K5, K14 and K19 were assayed and keratinocyte purity was nearly 100%.

CONCLUSIONA novel, simple and effective method can be used to rapidly obtain highly purified keratinocytes from normal human cervical epithelium.

Cell Culture Techniques ; methods ; Cervix Uteri ; cytology ; Epithelial Cells ; cytology ; Female ; Humans ; Keratinocytes ; cytology

OBJECTIVETo investigate the sex chromosomes and the expression of insulin-like growth factor-II (IGF-II) on activated human unfertilized oocytes after intracytoplasmic sperm injection(ICSI) with calcium ionophore A23187 and puromycin.

METHODSAll 95 discarded oocyes that showed no evidence of fertilization at 16-18 h after in vitro maturation and intracytoplasmic sperm injection cycles (IVM-ICSI)/conventional ICSI were exposed to calcium ionophore A23187 (5 micromol/L) for 5 min and then were incubated with puromycin (10 microg/mL) for 4 h. After activation, the oocytes were cultured in vitro for 3-5 days. The sex chromosome analysis was performed by dual color fluorescence in situ hybridization. The expression of IGF-II on the activated embryos, normal embryos, and parthenotes was examined.

RESULTSThe combination of calcium ionophore A23187 with puromycin could activate the unfertilized oocytes 22 h after ICSI. The activated rate, cleavage rate, and quality of activated embryos of the IVM-ICSI group were similar to those of ICSI group, respectively. Sex chromosome analysis indicated that 8 male and 5 female embryos had been derived from two pronucleus and a second polar body. The expression of IGF-II on activated embryos and normal embryos was high and similar, which was much stronger than that of parthenotes.

CONCLUSIONThe combination of calcium ionophore A23187 with puromycin could effectively activate unfertilized oocytes 22 h after ICSI. Moreover, the unfertilized oocytes activated by calcium ionophore A23187 and puromycin had normal sex chromosomes and expression of IGF-II like the normal embryos. These suggest that oocyte activation may be considered as a remedial measure in the presence of total or nearly total fertilization failure in ICSI.

Calcimycin ; pharmacology ; Chromosomes, Human, X ; genetics ; Chromosomes, Human, Y ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Insulin-Like Growth Factor II ; metabolism ; Ionophores ; pharmacology ; Oocytes ; cytology ; drug effects ; metabolism ; Puromycin ; pharmacology ; Sperm Injections, Intracytoplasmic ; methods