The present study investigated the relationship between DNA-dependent protein kinase (DNA-PK) and radiosensitivity of nasopharyngeal carcinoma (NPC) cell lines. The dose-survival relationship for NPC cell lines, CNE1 and CNE2, was analyzed using clonogenic formation assay, the activity of DNA-PK of the two cell lines was measured using the Signa TECT DNA-PK assay kit, and the localization and expression of Kus (a heterodimer) and DNA-PKcs protein in CNE1 and CNE2 before irradiation and 15 min, 1 h, 6 h, 12 h, 24 h after 4 Gy irradiation were analyzed by immunofluorescence, laser scanning confocal microscope (LSCM) and Western blot. The results showed that the surviving fraction of CNE1 was higher than that of CNE2 at each dose. The DNA-PK activity of CNE1 was also significantly higher than that of CNE2 before and after irradiation (P<0.05), while the expression of total Ku70/Ku80 in CNE1 and CNE2 had no significant difference. Increasing translocation of Ku70 and Ku80 from the cytoplasm to the nuclei in the two cell lines was observed with increase of irradiation time as detected by Western blot, and the immunofluorescence of the DNA-PK complex subunits showed greater nuclear translocation in CNE1 than CNE2 after irradiation. The results suggest that the relatively higher radio-resistance of CNE1 correlates with the higher activity of DNA-PK as compared to that of more radiosensitive CNE2 (or lower radio-resistance) before and after irradiation. Thus, DNA-PK activity may be a useful predictor of radiosensitivity of NPC.
Carcinoma ; Cell Line, Tumor ; enzymology ; radiation effects ; DNA-Activated Protein Kinase ; metabolism ; Humans ; Nasopharyngeal Neoplasms ; enzymology ; Radiation Tolerance

BACKGROUNDThe prevalence of malocclusion in modern population is higher than that in the excavated samples from the ancient times. Presently, the prevalence of juvenile malocclusion in the early stage of permanent teeth is as high as 72.92% in China. This study aimed to observe and evaluate the prevalence and severity of malocclusions in a sample of Xia Dynasty in China, and to compare these findings with the modern Chinese population.

METHODSThe material consisted of 38 male and 18 female protohistoric skulls of Xia Dynasty 4000 years ago. Of 86 dental arches, 29 cases had the jaw relationships. Tooth crowding, diastema, individual tooth malposition and malocclusion were studied.

RESULTSOf the samples, 23.3% showed tooth alignment problems including crowding (8.1%), diastema (9.3%), and individual tooth malposition (5.8%). The prevalence of malocclusion was 27.6%, mainly presented as Angle Class I.

CONCLUSIONSIt is indicated that over thousands of years from Neolithic Age (6000 - 7000 years ago) to Xia Dynasty (4000 years ago), the prevalence of malocclusion did not change significantly. The prevalence of malocclusion of Xia Dynasty samples was much lower than that of modern population.

China ; epidemiology ; Diastema ; Female ; History, Ancient ; Humans ; Male ; Malocclusion ; epidemiology ; history

OBJECTIVETo explore the effect of the improved plastic and reconstruction of the anus in situ.

METHODSImproved plastic and reconstruction of anus in situ was performed in 38 cases of low rectal cancers operated while Miles radical operation. Improvement includes: (1) The internal sphincter was rebuilt with 4 layers of muscle layer of the endmost of colon. (2) The last of gracilis was divided into 2 parts to reconstruct the superficial part and deep part of external sphincter muscle. (3) The rectum cape improvement is to firmly stitch the levator ani outside the external sphincter muscle in front of the colon. (4) The rectum valve is improved into three artificial rectum valves.

RESULTSThe form and function and their long term survival rate were good, the rate of superior anus function was 94.73%.

CONCLUSIONIt mains the results of improved plastic and reconstruction of anus in situ is near that of normal persons.

Adult ; Aged ; Anal Canal ; surgery ; Female ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Rectum ; surgery

OBJECTIVETo evaluate the feasibility, toxicity and tumor control of intensity modulated radiation therapy (IMRT) for recurrent nasopharyngeal carcinoma.

METHODSFourty-nine patients (Karnofsky performance status (KPS) >or= 80) with local-regional recurrence in the nasopharynx were treated with full course IMRT. Three patients with cervical lymph node metastasis (N1 2 and N3 1) were further supplemented with 5 to 6 courses of chemotherapy (Cisplatin + 5-Fu) after IMRT.

RESULTSThe results of treatment plan showed that the mean dose of covering gross tumor volume (GTV) (D(95)) in the nasopharynx was 68.09 Gy and the mean volume of GTV (V(95)) receiving the 95% dose was 98.46%. The mean dose of GTV, clinical target volume CTV1 and CTV2 in the targets were 71.40 Gy, 63.63 Gy and 59.81 Gy. The median follow-up time was 9 months (range 3 to 16 months). The local-regional progression-free survival was 100% with local-regional residual disease in 3 (6.1%) cases but was complicated with nasopharyngeal mucosa necrosis in 14 (28.6%) cases after IMRT.

CONCLUSIONIntensity modulated radiation therapy, as a re-treatment option for recurrent nasopharyngeal carcinoma, is able to improve the tumor target coverage and spare the adjacent critical structures. As high dose IMRT can result in radio-necrosis of nasopharyngeal mucosa, the prescription dose of GTV should be suitably decreased to 60 - 65 Gy.

Adult ; Aged ; Carcinoma, Squamous Cell ; pathology ; radiotherapy ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; pathology ; radiotherapy ; Neoplasm Recurrence, Local ; radiotherapy ; Neoplasm Staging ; Radiation Injuries ; pathology ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; Radiotherapy, Conformal ; methods

OBJECTIVETo establish a favorable formula of estimation adult standard liver volume (SLV) based on the data of living donors for liver transplantation.

METHODSFrom March 2005 to December 2007, 90 Adult-to-Adult living donor liver transplantation were performed. The donors' anthropometric data of gender, year, body weight and body height was collected prospectively. The total liver volume (TLV) of 90 living donors was measured by computed tomography. The correlation between TLV and several factors including body weight index (BMI) and body surface area (BSA) were analyzed. Then multiple stepwise linear regression analysis was performed and a new equation predicting SLV to improve approximate TLV was determined. TLV was then compared with the estimation standard liver volume (ESLV) calculated using our formula and published formulas in literature previously.

RESULTSAll the subjects had a mean body weight of (62.4 +/- 8.7) kg. The mean total liver volume was (1319.1 +/- 167.0) ml. There were positively correlated between TLV and body weight (BW), which could be expressed in the equation: TLV (ml) = 12.5 x BW (kg) + 536.4 (r(2) = 0.43, P < 0.01). Compared TLV with ELSV, which were calculated using previously published formulas based on the anthropometric data of the 90 subjects, the difference was be statistically significant.

CONCLUSIONA new simple formula is established that might be a more suitable to calculate TLV in Chinese adults.

Adult ; Female ; Humans ; Linear Models ; Liver ; anatomy & histology ; diagnostic imaging ; Liver Transplantation ; Living Donors ; Male ; Middle Aged ; Organ Size ; Tomography, X-Ray Computed

OBJECTIVETo investigate the changes in brain injury after the induction chemotherapy in children with acute lymphoblastic leukemia (ALL) by cranial MRI.

METHODSThe clinical data and cranial MRI results of 62 children with ALL who were hospitalized from March 2014 to June 2015 were analyzed retrospectively.

RESULTSBefore chemotherapy, MRI showed bone marrow infiltration of the skull in 33 patients (53%); the children with WBC<20×10(9)/Lhad a significantly lower incidence rate of bone marrow infiltration of the skull than those with WBC≥20×10(9)/L (16 patients/42% vs 17 patients/71%; P<0.05), and the high-risk group had a significantly higher incidence rate of bone marrow infiltration of the skull than the non-high-risk group (71% vs 44%; P<0.05). Before chemotherapy, there were 4 cases (7%) of brain atrophy, and 2 cases (3%) of abnormal signals in the sensory conduction bundle. MRI reexamination in 28 patients after 3 months of chemotherapy showed 3 new cases (11%) of brain atrophy and 1 aggravated case of brain atrophy.

CONCLUSIONSThe children with ALL have bone marrow infiltration of the skull, brain atrophy, and abnormal signals in the sensory conduction bundle before chemotherapy, especially bone marrow infiltration of the skull, and some changes in brain injury disappear after treatment.

Adolescent ; Bone Marrow ; pathology ; Brain ; drug effects ; pathology ; Child ; Child, Preschool ; Female ; Humans ; Induction Chemotherapy ; adverse effects ; Infant ; Magnetic Resonance Imaging ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; pathology ; Retrospective Studies ; Skull ; pathology

OBJECTIVETo investigate the correlations of serum interleukin-18 (IL-18) level and IL-18 gene promoter polymorphisms to the development of sepsis in children.

METHODUsing enzyme-linked immunosorbent assay (ELISA), the authors tested the serum IL-18 level in 90 patients with sepsis and 123 normal controls, and their single nucleotide polymorphisms of the promoter region of IL-18 gene at position -607C/A and -137G/C were detected using polymerase chain reaction with sequence specific primers method and sequencing technique.

RESULT(1) The serum IL-18 level in sepsis groups was (196.56 +/- 157.32) pg/ml that was significantly higher than (66.16 +/- 41.63) pg/ml in normal controls (P < 0.01), the more severe the degree of sepsis was, the more significantly higher the serum IL-18 level was. The serum IL-18 level in non serious sepsis group was (152.87 +/- 114.96) pg/ml that was significantly higher than (66.16 +/- 41.63) pg/ml in normal controls, the serum IL-18 level in serious sepsis group was (191.98 +/- 169.72) pg/ml that was significantly higher than that in non serious sepsis group, and the serum IL-18 level in extremely serious sepsis patients was (323.89 +/- 159.35) pg/ml, the difference was highly significant (P = 0.000). The difference was significant among the groups with different severity of sepsis (P < 0.01). There was a negative correlation between PCIS (pediatric critical illness score) of sepsis and the serum IL-18 level (P < 0.01). (2) There were polymorphisms in IL-18 gene promoter of matched healthy children and sepsis in children. The GG genotype frequency (61.8%) of IL-18-137G/C in healthy children was the highest, followed by GC genotype (35.8%) and CC genotype (2.4%) in sequence. The G allele frequency (79.7%) was higher in IL-18-137G/C of healthy children than C allele (20.3%). The GG genotype frequency (71.1%) of IL-18-137G/C in septic children was the highest, the next were GC genotype (26.7%) and CC genotype (2.2%). The G allele frequency (84.4%) was higher in IL-18-137G/C of septic children than C allele (15.6%). The CA genotype frequency (61.0%) of IL-18-607C/A in healthy children was the highest, followed by CC genotype (26.8%) and AA genotype (12.2%). The C allele frequency (57.3%) was higher in IL-18-607C/A of healthy children than A allele (42.7%). The CA genotype frequency (76.7%) of IL-18-607C/A in septic children was the highest, followed by CC genotype (21.1%) and AA genotype (2.2%) in sequence. The C allele frequency (59.4%) was higher in IL-18-607C/A of septic children than A allele (40.6%). (3) The genotype frequency of IL-18-607 CA was 76.7% in sepsis groups that was significantly higher than 61.0% in normal controls, and the genotype frequency of -607 AA was 2.2% in sepsis groups that was significantly lower than 12.2% in normal controls, the difference was significant (P < 0.05). (4) In the order of -137CC, -137GC, -137GG, the serum IL-18 level in normal controls were as follows: (45.67 +/- 28.36) pg/ml, (53.27 +/- 37.91) pg/ml, (76.91 +/- 42.44) pg/ml, and with (140.50 +/- 60.10) pg/ml, (184.42 +/- 157.33) pg/ml, (237.02 +/- 161.76) pg/ml respectively in sepsis groups. In the order of -607AA, -607CA, -607CC, the serum IL-18 level in normal controls were: (48.80 +/- 32.11) pg/ml, (68.41 +/- 42.53) pg/ml, (70.17 +/- 43.87) pg/ml; and with (141.50 +/- 64.35) pg/ml, (151.21 +/- 121.19) pg/ml, (211.16 +/- 163.64) pg/ml respectively in sepsis groups. The difference was not significant among different groups (P > 0.05).

CONCLUSIONThe serum IL-18 level in sepsis groups was significantly higher than that in normal controls, which was related to the severity of sepsis. It was possible that the genotype of -607CA carriers was susceptible to sepsis, which mean that the genotype of -607CA might be susceptible genotype of sepsis. However, the genotype of -607AA might play an oppose role in the risk of sepsis.

Adolescent ; Child ; Child, Preschool ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Infant ; Interleukin-18 ; blood ; genetics ; Male ; Polymorphism, Genetic ; Promoter Regions, Genetic ; Sepsis ; blood ; genetics

OBJECTIVETo clone the recombinant human islet neogenesis-associated protein (rhINGAP) gene for its secretory expression in Pichia pastoris.

METHODSINGAP gene was amplified with PCR and inserted between Xho I and EcoR I downstream sites of the alpha factor of the recombinant plasmid alpha/pUC18. The fusion gene of alpha factor and INGAP was subsequently inserted between BamH I and EcoR I sites of the plasmid pPIC9K of P. pastoris. After confirmation with restriction enzyme digestion and sequencing, the positive recombinant plasmid that integrated INGAP gene was linearized with Sal I digestion and transformed into the yeast host strain GS115 through electroporation. The yeast transformants that harbored the INGAP gene with high copies were selected with the auxotroph medium and G418, followed then by PCR verification of the positive transformants, from which the expression of recombinant human INGAP was induced with methanol as the only carbone source. The antigenic activity of the desired protein was then detected using Western blotting and enzyme-linked immunosorbent assay (ELISA).

RESULTS AND CONCLUSIONThe recombinant expression plasmid INGAP/pPIC9K was successfully constructed, and 3 positive transformants were obtained. The expressed protein showed good antigenic activity as confirmed by Western blotting and ELISA.

Antigens, Neoplasm ; genetics ; metabolism ; Biomarkers, Tumor ; genetics ; metabolism ; Blotting, Western ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Humans ; Islets of Langerhans ; metabolism ; Lectins, C-Type ; genetics ; metabolism ; Pancreatitis-Associated Proteins ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; metabolism

OBJECTIVETo explore the feasibility of applying autosomal single nucleotide polymorphisms (SNPs) on parentage testing.

METHODSAll SNP genotyping results of HapMap (r27) were downloaded from the website. With self-made computer programs, SNPs were extracted when their minor allele frequency (MAF) were ≥ 0.30 among all of the 11 HapMap populations. Ninety-six SNPs were chosen and integrated into the Illumina Goldengate bead arrays on the condition that no linkage disequilibrium was found between them. Three father-child-mother trios (9 samples in total) were tested with the arrays. Cumulative paternity index (CPI) was then calculated and compared with genotyping results using 15 short tandem repeats (STRs)(Identifiler(TM)).

RESULTSFamily 1 was found to have nine SNPs or seven STRs that did not conform to the Mendelian laws, Family 2 had 13 such SNPs or seven STRs, and Family 3 only had one such SNP but no STR. For Family 3, when all of the 96 SNPs were used in combine, the CPI was 1207, which had contrasted with the CPI by the 15 STRs, i.e., 355 869.

CONCLUSIONWhen applied to paternity testing, the paternity exclusion (PE) value for a SNP is usually less than 1/3 of that of a STR. The proportion of SNPs not comforming to the Mendelian laws for the tested SNPs may not be as high as that of inconsistent STRs over all tested STRs. Because of the low mutation rate of a SNP, the CPI will be greatly reduced even if one SNP did not conform to the Mendelian laws. Therefore, highly accurate testing methods are required to reduce artificial errors when applying SNPs for paternity testing.

Fathers ; Female ; Genetic Testing ; methods ; Genotype ; HapMap Project ; Humans ; Male ; Mothers ; Paternity ; Polymorphism, Single Nucleotide ; genetics

OBJECTIVEThrough monitoring esophageal dynamic change, and detection of laryngopharyngeal reflux(LPR) and gastroesophageal reflux events,to discuss the relationship of vocal cord polyps with laryngopharyngeal reflux.

METHODSThirty-two patients with vocal cord polyps were diagnosed by electronic laryngoscopy in Nanfang Hospital between October 2011 to May 2012. This study applied high-resolution esophageal manometry (HRM) and ambulatory 24-hour multichannel intraluminal impedance-pH monitoring (MII-pH) to obtain the upper esophageal sphincter(UES) and lower esophageal sphincter pressure, characteristics of sectional esophageal motility; laryngopharyngeal reflux (LPR)and gastroesophageal reflux events, as well as the reflux properties of substances. Sixteen healthy volunteers were recruited as normal controls.

RESULTSUES relaxation duration, duration of UES relaxation time, UES relaxation recovery time and mean length of LES were all shorter than those of the control group (t were 2.244, 2.624, 2.310 and -2.397, P < 0.05). There were 40.6% (13/32) LPR and 50.0% (16/32) gastroesophageal reflux found in vocal polyp patients. Median number (M [P25; P75]) of laryngopharyngeal acid reflux events were 0.5[0.0;3.5] and 0.0[0.0;0.0] in vocal polyp group and the controls, median mean time of laryngopharyngeal acid exposure 0.1[0.0;1.7] and 0.0[0.0;0.0] min, median clearance time of laryngopharyngeal acid were 3.5[0.0;53.5] and 0.0[0.0;0.0] s, median scores of DeMeester were 14.8[1.6;31.3] and 1.8[1.1;4.1] and median frequency of total liquid reflux episodes were 46.5[25.3;69.0] and 32.5[20.0;36.3], respectively. The median numbers of laryngopharyngeal acid reflux events, time of acid exposure, time of acid clearance, DeMeester scores and frequency of total liquid reflux episodes were increased or higher in vocal polyp group than those in the controls (z were 2.481, 2.767, 2.767, 2.344 and 1.980, P < 0.05).

CONCLUSIONSThere are upper esophageal sphincter and Lower esophageal sphincter dismotility in vocal polyp patients with LPR. LPR events were dominated by acid reflux in upright position.Esophageal dynamic disfunction and LPR should be considered in the study of the pathogenesis of vocal cords polyps.

Electric Impedance ; Esophageal Sphincter, Lower ; Esophageal pH Monitoring ; Gastroesophageal Reflux ; diagnosis ; Humans ; Laryngeal Diseases ; diagnosis ; Laryngopharyngeal Reflux ; diagnosis ; Laryngoscopy ; Manometry ; Polyps ; diagnosis ; Vocal Cords