Objective To investigate and evaluate the polymorphism distribution of arsenic(+3 oxidation state)methyhransferase(AS3MT)5'-UTR VNTR in Chinese populations.Methods Genomic DNA was extracted from peripheral blood anti-coagulated with ACD of 1440 individuals in a standard phenol-chloroform protocol.The phenotypes of AS3MT 5'-UTR VNTR were determined by polymerase chain reaction(PCR)associated with agarose gel electrophoresis.Results Of the 1440 individuals,771(53.5%),426(29.6%),211(14.7%),16(1.1%)and 16(1.1%)were carriers of the V2/V3(AB/A2B),V3/V3(A2B/A2B),V2/V2(AB/AB),V2/V4(AB/A3B)and V3/V4(A2B/A3B)genotype,respectively.The AB(V2),A2B(V3)and A3B(V4)allele frequency was 41.9%,57.0%,1.1%respectively.The differences of AB(V2)and A2B(V3)allele frequency were all significant between the northern and southern populations respectively(χ2=23.39,χ2=33.28,P＜0.007).Conclusions In different regions the AB(V2)and A2B(V3)allele frequency is different,the AS3MT 5'-UTR VNTR polymorphism can be used to evaluate the susceptivity of arsenieosis.

Objective To investigate subacute exposure of airborne particulate matter (PM) on pregnancy and fetal development in female mice. Methods Forty female and forty male ICR adult mice group (A), small (B) , middle (C) , large (D) or overdose (E) PM challenge groups (n = 8 - 11), and were administered with 30 μl of phosphate buffered solution (A) or resuspended standard PM SRM 1649a at 0.09 (B), 0.52 (C), 1.85 (D) or 69.2 (E) μg/μl, once per trid from d 0 till d 19 of pregnancy via instillation onto the base of the tongue. Fetal mice were harvested by cesarean section at the time when spontaneous delivery occurred. Body weight of the pregnant mice, gestational days, intrauterine survival and growth, hepatic and pneumonic histopathological changes of the fetal mice were investigated. Lung/body and liver/body weight ratios were calculated. Expressions of mRNA and protein of CYP1A1 in the fetal lung and CYP1 A2 in the fetal liver were assayed. Results (1) All of the pregnant mice survived pregnancy throughout the entire experiment. Body weight of the pregnant mice was not significantly different among all the groups at gestational d 1 and 7 (P ＞ 0.05), but significantly lower in group E [(41.8 ± 5.8) and (48.9 ± 8.9) g] than in group A [(45.9 ± 1.8) and (56.2 ± 4.9) g] at gestational d 14 and 18 (P ＜0.05). The gestational days were significantly decreased in group E [(19.3 ± 1.3) d] when compared with group A [(20.5 ± 0.7) d; P ＜ 0.05] and were not significantly different among groups A, B, C and D (P ＞ 0.05). Lung/body and liver/body weight ratios of the fetal mice were significantly increased in group E [(1.21 ±0.18) and (4.68 ±0.21)%] as compared with groups A, B, C and D (P＜0.05). (2)Mortality rates of the fetuses were significantly higher in group E (23.0%) than in groups A (0.8%), B (0.9%), C (1.7%) and D (3.7%) (P ＜ 0.05), but were not significantly different among groups A,B, C and D (P ＞ 0.05) despite of an increasing tendency. (3) Pathological changes in the liver and lung of the fetuses were conspicuous in group E. The fetal liver injury was histopathologically evidenced by deranged tissue structure, degenerated parenchyma of hepatic cells, and mildly stained cytoplasm. Adipose degeneration was represented by clear-boundary intracytoplasmic vacuoles in most of the liver cells, and cell pyknosis with heavily stained cytoplasm was observed in some of the liver cells. Inflammatory cell infiltration and focal necrosis were occasionally found in the hepatic tissue. The fetal lung exhibited bronchiole with narrow lumina, vascular engorgement in the submucosal layer, interstitial and alveolar edema, thickened alveolar septum, granulocyte and lymphocyte infiltrations within the pulmonary alveoli and around the bronchioles. The above pathological changes were lesser in groups C and D, and were not or least found in groups A and B. (4) Protein expressions of CYP1A1 in the fetal lung and CYP1A2 in the fetal liver were significantly increased in group E (1.20 ± 0.40 and 2.55 ± 0.89) when compared with group A (0.77 ±0.36 and 2.08 ±0.31) (P ＜ 0.05). mRNA expressions of CYP1A1 in the fetal lung were significantly increased in groups C (0.36 ±0.12), D (0.41 ±0.08) and E (0.43 ±0.11) compared with group A (0.21 ±0.10), and significantly increased in groups D and E compared with group B (0.28 ±0.10,P＜0.05). mRNA expressions of CYP1 A2 in the fetal liver were significantly increased in groups C (0.37 ±0.13), D (0.36 ±0.14) and E (0.43 ±0.16) compared with group A (0.21 ±0.03), and significantly increased in group E compared with group B (0.24± 0.11, P ＜ 0.05). Conclusions PM elicited embryotoxigenicity and resulted in adverse pregnancy outcomes in mice by intrauterine exposure of overdose PM. The expressions of cancer-related genes CYP1A1 and CYP1A2 were up-regulated in organs after the middle- and large-dose subacute exposure of PM, which may have a potential role on the future development.

Adenoma, Villous ; metabolism ; pathology ; surgery ; Aged ; Diagnosis, Differential ; Female ; Humans ; Keratin-20 ; metabolism ; Keratin-7 ; metabolism ; Mullerian Ducts ; pathology ; Papilloma ; pathology ; Vaginal Neoplasms ; metabolism ; pathology ; surgery

OBJECTIVETo identify the Cynomorii Herba and its analogues species using DNA barcoding technique.

METHODTotal genomic DNA extracted from all materials using the DNA extraction kit. The internal transcribed spacer 2 (ITS2) regions were amplified using polymerase chain reaction (PCR), and purified PCR products were sequenced bi-directionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner 3.7.1. The Kimura 2-Parameter (K2P) distances and GC content were computed using MEGA 5. 0. Species identification analyses were conducted through the species identification system for traditional Chinese medicine and neighbor-joining (NJ) trees.

RESULTThe ITS2 sequence lengths of Cynomorii Herba were 229 bp. The average intra-specific genetic distances of Cynomorii Herba were 0.003. The average inter-specific genetic distances between Cynomorii Herba and its adulterants species were 0.760. The results showed that the minimum inter-specific divergence is larger than the maximum intra-specific divergence. The species identification system for traditional Chinese medicine and NJ trees results indicated that Cynomorii Herba and its adulterants species can be easily identification.

CONCLUSIONThe ITS2 region is an efficient barcode for identification of Cynomorii Herba, which provide a new technique to ensure clinical safety in utilization of traditional Chinese medicine.

Cynomorium ; classification ; genetics ; DNA Barcoding, Taxonomic ; DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Polymerase Chain Reaction

Objective: To compare the prompt fixation strength and antifatigue strength of the locking and non-locking anterior cervical plating systems. Methods:AO Cervical Spine Locking Plate (CSLP), Danek Orion plate and AcroMed Acroplate were used on the lamb cervical spines. The CSLP and Orion were tested with screws locked and unlocked, and the Acroplate with unicortical and bicortical purchase. The fixation strength and pull-off strength of the screw-plate constructs were performed initially and after fatigue. Results: Locked CSLP and Orion constructs were more rigid than all unlocked unicortical systems initially and after cyclic loading. After fatigue testing, the strength of all unlocked constructs decreased significantly. There was no significant difference in pull-off strength between the CSLP, the Orion and the unicortical Acroplate. Conclusion: The locking mechanism significantly increases the prompt fixation strength and antifatigue strength of the tested screw-plate systems.

Objective: To investigate the long-term outcome of anterior decompression and bone graft fusion for cervical spondylotic myelopathy(CSM) and factors affecting the outcome. Methods: Two hundred and forty-five patients with CSM were treated with anterior cervical decompression and auto iliac bone graft fusion, of whom 31 had a second operation between 4 months and 2 years after operation. Follow-up studies were carried out within 5 to 15 years after operation, averaging 6.8 years. Results: Function evaluation: excellent in 118 cases (48.16%), good in 71 (28.98%), passable in 35 (14.29%) and poor in 21 (8.57%). According to the 40 points score method, there was an average of 8 point increase in all cases, of which 101 were between 36 to 40 points, 54 between 31 to 35 points. Conclusion: The long-term outcome of surgical treatment for CSM is definite. Significant factors affecting the outcome include timing of operation, degree of pathology and technique of surgery.

Adult ; Aged ; Carcinoma, Renal Cell ; complications ; mortality ; Humans ; Kidney Neoplasms ; complications ; mortality ; Male ; Neoplastic Cells, Circulating ; Prognosis ; Thrombosis ; etiology ; Vena Cava, Inferior

BACKGROUNDLung cancer is the leading cause of cancer-related death in China. Mutation analysis reveals that LKB1 inactivation is present in 30% of non-small-cell lung cancer (NSCLC), indicating its role as a tumor suppressor. However, the molecular mechanism is still not clear. Our study attempted to establish LKB1 stable knockdown NSCLC cell line, detect alterations in gene expression and identify the genes regulated by LKB1.

METHODSLKB1 stable knockdown H1299 cell line was established using a lentiviral short hairpin RNA. To identify the knockdown effect, LKB1 mRNA and protein expression level were evaluated with quantitative real-time PCR and Western blotting. We treated the cell lines with 2-deoxyglucose to determine if LKB1 protein function was impacted. Gene microarray analysis was performed to detect the gene expression alterations in LKB1 stable knockdown H1299 cells.

RESULTSLKB1 mRNA and protein expression were significantly suppressed in LKB1 stable knockdown H1299 cell line. 2-DG treatment had little impact on the phosphorylation of AMPK, which is the downstream target of LKB1, indicating the loss of function of LKB1. The microarray data showed that LKB1 knockdown resulted in expression alterations of 1243 kinds of genes, including those involved in cell migration, cell proliferation and cell apoptosis.

CONCLUSIONSThe establishment of LKB1 stable knockdown H1299 cell line provides us with a great tool to investigate various genes regulated by LKB1 through microarray. The discovery of cell proliferation and migration-related genes regulated by LKB1 is critical for unraveling molecular mechanisms of LKB1's role in the development and metastasis of lung cancer.

Blotting, Western ; Cell Line, Tumor ; Gene Expression Profiling ; methods ; Humans ; Lung Neoplasms ; genetics ; metabolism ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction

OBJECTIVETo study significance and limitations of the ratio of free to total prostate specific antigen (f/t PSA) in differential diagnosis between prostate cancer and benign prostatehyperplasia (BPH) with total PSA (tPSA) levels between 4 and 10 ng/ml.

METHODSWe analysed retrospectively 180 prostate cancer and BPH patients who were diagnosed and treated in our hospital from October 1998 to October 2002 and had serum tPSA levels between 4 and 10 ng/ml. Of the 180 patients, 36 (20%) were histologically confirmed as prostate cancer and 144 (80%) BPH. The tPSA and free PSA (fPSA) in serum were measured by micropartical enzyme immunoassay. Prostate volume was measured by transabdominal ultrasonography. We chose Student's t-test for comparison between prostate cancer and BPH groups. The correlation between prostate volume and f/t PSA was analyzed using Pearson's correlation coefficient.

RESULTSThe mean values of tPSA and f/t PSA were 6.75 ng/ml and 0.17 in patients with prostate cancer, 6.48 ng/ml and 0.25 in patients with BPH. The mean value of tPSA wasn't significantly different between patients with prostate cancer and BPH (P > 0.05). However, the mean value of f/t PSA of patients with prostate cancer was significantly lower than that of patients with BPH (P < 0.01). Furthermore, there were significant and positive correlation between prostate volume and f/t PSA in both groups with prostate cancer and BPH (prostate cancer group's correlation coefficient (r = 0.50, P < 0.01); BPH group (r = 0.24, P < 0.01). There was significant difference in f/t PSA between prostate cancer and BPH patients with prostate volumes more than 40 cm(3) (P < 0.05), but not between these two groups with prostate volumes more than 40 cm(3) (P > 0.05).

CONCLUSIONThe f/t PSA is significant in differential diagnosis between prostate cancer and BPH with tPSA levels between 4 and 10 ng/ml. But prostate volume has an effect on f/tPSA. The f/tPSA has diagnostic value of differentiation only when the prostate volume is less than 40 cm(3).

Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; Diagnosis, Differential ; Humans ; Kidney ; diagnostic imaging ; Male ; Middle Aged ; Prostate-Specific Antigen ; blood ; Prostatic Hyperplasia ; diagnosis ; diagnostic imaging ; Prostatic Neoplasms ; diagnosis ; diagnostic imaging ; Retrospective Studies ; Sensitivity and Specificity ; Ultrasonography