The regulatory region and the signal peptide sequence of the sacB gene has been amplified by PCR using Bacillus subtilis chromosomal DNA as template, and an inducible secretion vector has been developed based on this sequence, which was ligated with Bacillus subtilis alkaline proteinase gene. Transform Bacillus subtilis DB403 with this vector, and the expression of the inserted Bacillus subtilis alkaline proteinase gene can be induced by addition of sucrose into the medium.

OBJECTIVETo develop a human ovarian carcinoma SKOV3 model in severe combined immunodeficiency (SCID) mouse and to study its biologic characteristics.

METHODSHuman ovarian carcinoma SKOV3 cells were injected intraperitoneally into female SCID mouse to establish a transplantation model of human ovarian carcinoma. The biological characteristics, metastasis and morphology of transplanted tumors were studied.

RESULTAll tumors grew progressively with no sign of regression. The tumor cells spread around the peritoneal cavity and mainly on the diaphragm, mesentery, peritoneum and around the liver, which was confirmed by histopathology. The morphology, growth pattern and CA125 secretion of primary culture of transplanted cells remained as same as those of ovarian carcinoma cell line SKOV3.

CONCLUSIONAn intraperitoneal transplantation model of human ovarian carcinoma SKOV3 in SCID mice has been developed successfully, which can simulate the biological behavior of peritoneal metastasis of human ovarian carcinoma.

Animals ; Disease Models, Animal ; Female ; Humans ; Mice ; Mice, SCID ; Neoplasm Transplantation ; Ovarian Neoplasms ; pathology ; ultrastructure ; Peritoneal Neoplasms ; secondary ; Transplantation, Heterologous

BACKGROUNDPreoperative tumor grading becomes one of the most important predictors for lymphadenectomy at primary surgery for clinical stage I endometriod adenocarcinoma. However, there is an inconsistency of tumor grade between preoperative curettage and final hysterectomy specimens, and its associated factors are poorly understood. This study aimed to evaluate the accuracy of tumor grade by preoperative curettage so as to achieve a better stratified management for clinical stage I endometriod adenocarcinoma.

METHODSClinical data of totally 687 patients with clinical stage I endometriod adenocarcinoma who underwent preoperative curettage and primary surgery were retrospectively collected. Compared with final hysterectomy specimens, the sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of tumor grade by preoperative curettage were calculated and their associations with clinicopathologic parameters, including age, status of menopause, position of uterus, location and size of lesion, histological grade, depth of myometrial invasion, cervical invasion, extrauterine spread, peritoneal cytology, metastasis to retroperitoneal lymph node, serum CA125 level, and hormone receptor status, were analyzed.

RESULTSIn final hysterectomy specimens, 139 of 259 grade 1 patients by curettage were upgraded to grade 1 or 2; 31 of 296 grade 2 were upgraded to grade 3, with a significantly discrepant rate of 40.9% (281/687) and an upgraded rate of 24.7% (170/687). The specificity and negative predictive value for grade 3 were 90.7% and 89.9%, while the sensitivity and positive predictive value for grade 1 were 67.1% and 40.9%, respectively.

CONCLUSIONSPreoperative tumor grade by curettage does not accurately predict final histological results, especially in those classified as grade 1. Complete surgical staging seems to be necessary for clinical stage I endometriod adenocarcinoma.

Adenocarcinoma ; diagnosis ; pathology ; surgery ; Adult ; Curettage ; methods ; Endometrial Neoplasms ; diagnosis ; pathology ; surgery ; Female ; Humans ; Hysterectomy ; Middle Aged ; Neoplasm Staging ; methods ; Retrospective Studies

·AIM: To study the therapeutic method of XL -radiofrequency ablation skin apparatus (invented by Rehabilitation medicine, PLA General Hospital) in micro-resecting the lacrimal punctum neoplasm and to observe its clinical effects. ·METHODS:XL-radiofrequency ablation skin apparatus was performed on 7 cases (7 eye lesions) of lacrimal punctum neoplasm including 3 intradermal nevus, 1 boundary nevus, 1 mixed nevus with inflammatory granuloma,1 inflammatory granuloma,and 1 squamous papillary cell tumor by separating layer, point and flake removing methods. The clinical effect was investigated 2-6mo postoperatively and leftover was removed by twice. The postoperative follow- up durations ranged from 6 to 60mo. ·RESULTS: Totally 6 cases (6 eye lesions) were cured successfully by once but 1 case by twice. They were all without recurrence, cicatrix, palpebral margin deformity, obstruction of lacrimal punctum, stenosis of lacrimal punctum or epiphora. Micro - resecting the lacrimal punctum neoplasm using the XL-radiofrequency ablation skin apparatus had several advantages, such as high accuracy,no cicatrix,no bleeding,no epiphora,no damage to the structure of lacrimal canaliculus, short operation time, little pain during the procedure, without hospitalization, slight inflammation reaction after procedure. ·CONCLUSION: The efficacy of XL - radiofrequency ablation skin apparatus is significantly more evident and highly accurate in micro - resecting lacrimal punctum neoplasm.

To investigate the effects of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor simvastatin (SV) on proliferation, apoptosis and the PI3K/AKT signaling pathway in human acute monocytic leukemia cell line SHI-1. SHI-1 cells were incubated with different concentrations of SV (5, 10, 15 µmol/L). Otherwise, SHI-1 cells without any treatment were used as control. Cells in different groups were collected at 24, 48 and 72 h after incubation for further detection. MTT method was used to assay the growth inhibition rate and flow cytometry was used to detect the early stage apoptosis ratio. The human PI3K-AKT Signaling Pathway RT(2) Profiler(TM) PCR Array was used to detect the expression of 84 genes involved in PI3K-AKT signaling. The results indicated that the SV inhibited the proliferation and inducted the apoptosis of SHI-1 cells in time- and dose-dependent manners significantly. The growth inhibition rates of SHI-1 cells treated with 15 µmol/L SV for 24, 48 and 72 h were 26.82, 47.09 and 63.92, respectively; and their early stage apoptosis ratios were 5.75, 13.25 and 15.59, respectively. Compared with the control group, expression levels of 39 genes were changed in the group of 15 µmol/L SV at 48 h, among them 26 genes were down-regulated and 13 genes were up-regulated. It is concluded that the SV can inhibit proliferation and induce apoptosis of SHI-1 cells, and the mechanism may be associated with the changes of gene expression level in PI3K-AKT signaling pathway regulated by SV.
Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Down-Regulation ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia, Monocytic, Acute ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; drug effects ; Simvastatin ; pharmacology

Animals ; Female ; Liver ; enzymology ; Liver Cirrhosis, Experimental ; enzymology ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-2 ; biosynthesis ; genetics

OBJECTIVETo investigate neuroendocrine differentiation and its mechanism in ovarian epithelial tumors.

METHODSNeuroendocrine (NE) cells were identified by immunohistochemical staining for chromogranin A and synaptophysin in 79 cases of ovarian epithelial tumor and 22 cases of normal ovary. Double-labeling technique was used for simultaneous detection of CgA and epithelial membrane antigean (EMA), and the staining intensity was quantitatively evaluated using an image analysis system.

RESULTSThe positive staining rate for CgA and SYN in ovarian epithelial tumors was 59.4% and 65.36%, respectively, which was higher than that in normal ovary (P=0.000), in which numerous NE cells were found. Both the number and staining intensity of NE cells in ovarian epithelial tumor were increased as compared with normal ovary. Cells co-expressing CgA and EMA were detected in the ovarian epithelial tumors.

CONCLUSIONThe presence of NE cells in ovarian epithelial tumor suggests heterogeneity of the tumors, and the occurrence of "multidirectional differentiation cells" within the these tumors indicates that NE cells might derive from malignant cells with multidirectional differentiation capacity.

Adult ; Aged ; Case-Control Studies ; Cell Differentiation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Middle Aged ; Neoplasms, Glandular and Epithelial ; genetics ; metabolism ; pathology ; Neuroendocrine Cells ; metabolism ; pathology ; Ovarian Neoplasms ; genetics ; metabolism ; pathology ; Ovary ; cytology ; metabolism ; pathology ; Young Adult

Objective To compare the clinical efficacy and safety of levetiracetam tablet and carbamazepine tablet in the treatment of children's intractable epilepsy.Methods A total of 96 children with intractable epilepsy were randomly divided into control group and treatment group with 48 cases per group.Control group was given carbamazepine 4-8 mg · kg-1 · d-1,tid,oral.Treatment group was given levetiracetam 4 mg · kg-1,bid,the maximum dose was 16 mg · kg-1 at the speed as 4 mg · kg-1 with every 2 weeks.Two groups were treated for 8 months.The clinical efficacy,neurocognitive function test [verbal intelligence quotient(VIQ),performance intelligence quotient (PIQ),total intelligence quotient(TIQ) and short-term visual memory],and adverse drug reactions were compared between two groups.Results After treatment,the total effective rates in treatment and control groups were 87.50％ (42 cases/48 cases) and 79.17％ (38 cases/48 cases) with significant difference (P ＜ 0.05).After treatment,the main indexes in treatment and control groups were compared:VIQ were (106.97 ± 5.65) and (95.25 ± 3.28) points,PIQ were (116.45 ± 5.16) and (103.61 ± 2.74) points,TIQ were(119.92 ± 4.69) and(95.20 ± 3.24) points,short-term visual memory were (18.45 ± 2.17) and (13.84 ± 1.81) s,the differences were statistically significant (all P ＜ 0.05).The adverse drug reactions of two groups were based on emotional,drowsiness,palpitations and dizziness,also,the incidences of adverse drug reactions in treatment and control groups were 12.50％ and 16.67％ without significant difference (P ＞ 0.05).Conclusion Levetiracetam tablet has a definitive clinical efficacy in the treatment of children's intractable epilepsy,which is better than carbamazepine tablet.Levetiracetam tablet can improve the cognitive ability for children's intractable epilepsy,without increasing the incidence of adverse drug reactions.

OBJECTIVETo evaluate immunogenicity after primary vaccination by different sequential program of inactivated poliovirus vaccine (IPV) and oral poliovirus vaccine (OPV).

METHODSChildren of 2 months old (60-89 days) selected in Beijing were assigned to 4 groups, 1 dose IPV plus 2 doses OPV (I-O-O, 122 children), 2 doses IPV plus 1 dose OPV(I-I-O, 103 children), 3 doses IPV (I-I-I, 114 children), and 3 doses OPV (O-O-O, 106 children), and were vaccinated at the age of 2, 3, 4 months. Polio neutralizing antibody titers against poliovirus types 1, 2, and 3 were tested and protective rates were calculated before the 1st dose, after the last dose, and after the 1st and 2nd dose of IPV.

RESULTSAfter the primary immunization, geometric mean titers (GMT) of polio neutralizing antibody titers against poliovirus types 1, 2, and 3 were 788.32, 738.42 and 631.17 in O-O-O group, 212.02, 262.30 and 537.52 in I-I-I group, 940.35, 929.72 and 940.35 in I-O-O group and 901.09, 1102.68 and 1110.12 in I-I-O group (F values were 47.71, 53.84, and 9.81 respectively, all P values<0.01). The protective rate of three types among each group was 98.1% (104/106)-100.0% and the difference was not statistically significant (P>0.05). After the 1(st) dose of IPV, the GMT were 18.88, 37.77, 24.64 and the protective rate was 82.6% (122/138)-96.4% (133/138); after the 2nd dose of IPV, GMT were 177.03, 168.25, 321.86 and the protective rate was 99.1% (108/109)-100.0% (109/109) in antibody types 1, 2 and 3, respectively.

CONCLUSIONGMT of polio neutralizing antibody titers against poliovirus is higher after vaccination by sequential program of IPV and OPV than that by IPV or OPV 3-doses program. High level of protective rate after 2 doses of IPV in I-I-O group may lead to better protection from vaccine associated paralytic poliomyelitis (VAPP). Sequential program of IPV and OPV can be used to maintain high level of herd immunity and to prevent VAPP, and the I-I-O sequential program should be the first choice.

Humans ; Immunization Schedule ; Infant ; Poliovirus Vaccine, Inactivated ; administration & dosage ; immunology ; Poliovirus Vaccine, Oral ; administration & dosage ; immunology ; Vaccines, Attenuated ; immunology

Objective To evaluate the efficacy and safety of short-term treatment including fluoroquinolones anti-tuberculosis drugs for rifampicin resistant pulmonary tuberculosis(TB)in those areas carrying out the 'TB control project'.Methods TB cases involved in this study were from TB drug resistance surveillance in Heilongiiang province,Zhejiang province and Shenzhen city from 2004 to 2006.TB cases with rifampicin resistant were randomly divided into the treatment group(including fluoroquinolones anti-tuberculosis drugs group)and the control group(re-treatment regimen group).The treatment group was treated wim 3RFT AM ofx Pto PAS-INH/5RFT ofx Pto PAS.INH while the control group was treated with 3 H3R323E3S3/5 H3R3E3.Efficacy of short-term treatment was analyzed by per-protocol analysis(PP analysis)and intention-to-treat analysis(ITT analysis)while drug adverse reactions was also observed.Results (1)154 patients with rifampicin resistant pulmonary tuberculosis were recruited among them,25(16.2%)were only resistant to rifampicin,114(74.0%)to MDR-TB and 15(9.8%)to others(resistant R+S,resistant R+E and resistant R+E+S).114 TB cases completed the fuIl course of treatment,with 71 in the treatment group and 43 in the control group.(2)Sputum negative conversion rate of the treatment group and the control group were 78.9%and 65.1%(X2CMH=4.558,P=0.011)respectively,by per-protocol analysis.Sputum negative conversion rate of the treatment group and the control group were 65.9%and 40.6%(X2CMH=0.272,P=0.001)respectively,by intention-to-treat analysis.The sputum negative conversion rate of the treatment group was higher than in the control group when treating rifampicm resistant pulmonary tuberculosis and MDR-TB patients.(3)Three patients withdrew in each of the two groups because ofadverse effects to the drugs.Rates of adverse reaction to drugs appeared to be 23.9%(17/71)and 18.6%(8/43)in the treatment and in the control groups,with no statistically significant difference between the two groups.Conclusion The efficacy of treatment including fluoroquinolones anti-tuberculosis drugs group seemed beaer than the re-treatment regimen group in treating patients with rifampicin resistant pulmonary tuberculosis and those MDR-TB patients.