[Reference value of urine tangible component analyzer for healthy adults in Guangzhou area].

Objective: To establish the normal reference for iQ200 urine component analyzer of healthy adults in Guangzhou area. Methods: Five hundreds and four normal urine samples were screened from the healthy population with the normal dry chemistry results of urine and normal biochemical results of plasma, and the RBC, WBC and epithelial cells were determined by iQ200 urine component analyzer. Then 1 ml urine was accurately collected from the samples by centrifugation and the RBC, WBC and epithelial cells were counted with neubauer counter by two inspectors. The samples were divided into 4 groups according to the age (18-45 years and >45 years old) and sex (male and female). The counts of RBC, WBC and epithelial cell of 4 groups with iQ200 urine component analyzer and neubauer counter were statistically analyzed. Results: The difference was not significant in RBC, WBC and epithelial cells between the results of iQ200 urine component analyzer and the results of neubauer counter(all P>0.05). There was not significant difference of RBC, WBC and epithelial cells of the same sex and different age groups (all P>0.05). There was significant difference of RBC, WBC and epithelial cells between the different gender groups(all P<0.05). The 95% confidence interval reference range of iQ200 urine component analyzer were as follows: RBC 0-7 cells/µl (male), 0-8 cells/µl (female); WBC 1-6 cells/µl (male), 1-8 cells/µl (female); epithelial cells 0-2 cells/µl (male), 0-8 cells/µl (female). Conclusion: The reference of this research of iQ200 urine component analyzer will be useful for the laboratory and clinic, but still need the verification experiment before using in the laboratory.

[Relationship between phenotype and genotype of ABCB11 deficiency in siblings and literature review].

Objective: To explore the relationship between genotype and phenotype of ABCB11 deficiency. Methods: Clinical data of two siblings with ABCB11 deficiency were retrospectively analyzed. Related literature from PubMed, CNKI and Wangfang databases was reviewed to date (up to August 2017) with 'ABCB11 gene' or 'bile salt export pump', 'cholestasis' and 'child' as key words. Results: The patients were siblings. Both of them presented as jaundice, pruritus and hepatosplenomegaly since 3 days after birth. Significant laboratory findings on admission of the older sister included high total bilirubin, 170 µmol/L;conjugated bilirubin, 115.8 µmol/L;alanine aminotransferase, 168 U/L;total bile acid 186.3 µmol/L and normal gamma-glutamyl transpeptidase. While routine laboratory data of the younger brother were as follows: total bilirubin, 148.8 µmol/L;conjugated bilirubin, 96.3 µmol/L;alanine aminotransferase, 232.8 U/L;total bile acid 226 µmol/L, and normal gamma-glutamyl transpeptidase.Both received ursodeoxycholic acid and fat-soluble vitamins. Liver pathology of the younger brother showed giant hepatocytes with ballooning degeneration, focal necrosis and intrahepatic cholestasis. Both the patients harbor the same compound heterozygous mutations in ABCB11 gene, c.145C>T (p.Q49X) and c.1510G>A (p.E504K). The sister is 9 years old now, with normal liver function. Jaundice faded around 3 months after birth, pruritus relieved at age 5, and medications was stopped since then. The brother progressed to liver failure after an operation on perianal abscess when he was 8-month-old, and received living-related liver transplantation when he was 9 month and 20 days old (from his mother). Now he is 1 year and 5 months old, with normal liver function. Both are under our follow-up. Literature review revealed 18 ABCB11 deficiency patients from 7 families who had apparent different prognoses, within each family the siblings had the same ABCB11 gene mutation. Seven cases relieved after ursodeoxycholic acid therapy and/or partial external biliary diversion, 5 received orthotopic liver transplantation, 2 developed hepatocellular carcinoma and 4 cases died in childhood. Conclusions: The clinical manifestations of ABCB11 deficiency may vary greatly in patients carrying the same genotype, even in siblings. Patients should be managed in individualized maner.

[The relationship between the radioresistance of pancreatic cancer cell SW1990 and the induction of the Epithelial-Mesenchymal Transition: an in vitro study].

Objective: To investigate the relationship between the radioresistance of pancreatic cancer cell SW1990 and the induction of the Epithelial-Mesenchymal Transition (EMT). Methods: The radio-resistant pancreatic cancer cell SW1990-R were established by using the method of radiating pancreatic cancer cell SW1990 step by step and repeatedly. Then the changes of the morphology of the cell was observed by inverted phase contrast microscope, the radioresistance of SW1990-R was detected by colony-forming assay, and the apoptosis rate of the two cell lines after radiation were measured by flow cytometry. Then invasiveness and EMT-related genes was measured by trans-well test and qRT-PCR. Finally, the model of transplanted tumor on nude mouse was used to confirm the relationship between the radioresistance of pancreatic cancer cell SW1990 and the induction of EMT. Results: Compared with SW1990, SW1990-R had a lower radiosensitivity (survival fraction in 2 Gy, SF2: 0.326 3±0.007 3 vs 0.840 8±0.001 9, P<0.05) and lower apoptosis rate[(6.12±1.27) % vs (16.87±1.73)%, P<0.05]. Meanwhile, the invasive ability of SW1990-R were significant higher than that of SW1990 cell. According to the result of both in vivo and in vitro experiment, SW1990-R had a higher expression level of Vimentin and Snail, and lower expression level of E-cadherin when compared with SW1990. Conclusion: Compared with SW1990, the radio-resistant pancreatic cancer cell SW1990-R can induce the Epithelial-mesenchymal transition.

IGFBP-3 A-202C and C2133G polymorphisms and colorectal cancer risk: a meta-analysis of case-control studies.

Recent evidence suggests that genetic variations in the IGFBP-3 gene may impact susceptibility to colorectal cancer, but individually published results are inconclusive. Our meta-analysis was aimed at providing a more precise estimation of these associations. An extensive literature search was conducted for appropriate articles published before May 1, 2013. This meta-analysis was performed using the STATA 12.0 software. The crude odds ratios (OR) with 95% confidence interval (CI) were calculated. Eleven case-control studies were included with a total of 11,895 colorectal cancer patients and 17,147 healthy controls. Our meta-analysis indicated that the G variant of IGFBP-3 C2133G polymorphism may be associated with increased colorectal cancer risk. However, no statistically significant association was noted between IGFBP-3 A-202C polymorphism and colorectal cancer risk. No publication bias was detected in this meta-analysis. The current meta-analysis suggests that the IGFBP-3 C2133G polymorphism may confer susceptibility to colorectal cancer. The G variant of the IGFBP-3 C2133G polymorphism may serve as a useful biomarker for predicting the risk of colorectal cancer.

HIF1A gene Pro582Ser polymorphism and susceptibility to digestive tract cancers: a meta-analysis of case-control studies.

Many existing studies have demonstrated that common polymorphisms in the hypoxia inducible factor-1α (HIF-1A) may contribute to the development of digestive tract cancers, but individually published studies showed inconclusive results. This meta-analysis aimed to derive a precise estimation of the relationships between HIF1A Pro582Ser polymorphism and the risk of digestive tract cancers. We searched CISCOM, CINAHL, Web of Science, PubMed, Google Scholar, EBSCO, Cochrane Library, and CBM databases from inception through May 1, 2013. Meta-analysis was performed using the STATA 12.0 software. We assessed 6 case-control studies that included a total of 911 digestive tract cancer patients and 2774 healthy controls. Our meta-analysis indicated that HIF1A Pro582Ser polymorphism was associated with an increased risk of digestive tract cancer. Subgroup analysis by ethnicity suggested that HIF1A Pro582Ser polymorphism might increase an individual's susceptibility to digestive tract cancer in Asian populations. However, similar association was not observed in Caucasian populations. In conclusion, our findings suggest that HIF1A Pro582Ser polymorphism may contribute to the risk of digestive tract cancers, especially in Asian populations.

Circular closure is associated with the lowest rate of surgical site infection following stoma reversal: a systematic review and multiple treatment meta-analysis.

AIM: Stoma reversal is frequently complicated by surgical site infection (SSI). To reduce SSI, several techniques for skin closure have been studied, with no agreement on which is best. The aim of this study was to identify the skin closure technique associated with the lowest rate of SSI following stoma reversal. METHOD: We systematically searched MEDLINE (PubMed and OvidSP), Scopus and clinical registries from 1 January 1980 to 24 March 2012, and included original reports on adult patients following stoma reversal. A network of treatments was created to map the comparisons between skin closure techniques, including primary closure, primary closure with a drain, secondary closure, delayed primary closure, loose primary closure and circular closure. Pairwise meta-analyses were performed for all available direct comparisons of closure types and heterogeneity was assessed. A multiple-treatments meta-analysis was conducted to estimate relative treatment effects between competing closure types (reported as an odds ratio with 95% credible interval, and a probability that each treatment is best). Several sensitivity analyses were performed. RESULTS: Fifteen studies were identified with a total of 2921 cases of stoma reversal. Overall, study quality was poor with observed low (one study), moderate (seven studies) and high (seven studies) risk of bias. Circular closure was associated with the lowest SSI risk (OR 0.12; 95% CI 0.02-0.40) and was the best of six skin closure techniques (probability of being best = 68.9%). Circular closure remained the best after sensitivity analyses. CONCLUSION: This study showed that circular closure is the best skin closure technique after stoma reversal in terms of SSI rate, but the quality of supporting evidence is limited, precluding definite conclusions.

Isolation and identification of renal cell carcinoma-derived peptides associated with GP96.

We determined the possible associated determinants and analyzed whether gp96-_associated antigenic peptides can be found in renal cell carcinoma (RCC). The gp96-peptide complexes were chromatographically purified from resected tumor tissue of RCC patients. SDS-PAGE and Western blot analysis confirmed gp96 using the gp96 monoclonal antibody, and its concentration was measured using BCA. Approximately 20 to 50 µg gp96-peptide complexes was obtained from 1 g RCC tissue. The mass spectrometry (MS) analysis of the eluted peptides included the initial profiling using matrix-assisted laser desorption/ionization time-of-flight MS. Quadrupole time-of-flight MS combined with the Mascot search engine was used to identify the peptides and find proteins from primary sequence databases. MS analysis results demonstrated that the mass range of peptide associated with gp96 was from 1046.48 to 3501.56 Da. Further research confirmed the sequences of two gp96-associated peptides, namely, LVPLEGWGGNVM and PPVYYVPYVVL. However, the original protein of the two peptides could not be found. The results demonstrated that the gp96-associated peptides are small molecular peptides, and the two peptides are deduced to be RCC-associated peptides. The identified peptides were confirmed to be associated with gp96 using the protocols described above. However, the specificity and relevance of the association to the immunogenicity of gp96 remains to be examined. Further analysis must be accomplished before the findings can be applied in peptide vaccine.

Strategies to improve the killing of tumors using temozolomide: targeting the DNA repair protein MGMT.

Alkylating agents such as temozolomide (TMZ) are effective anticancer drugs for treating a variety of solid tumors including melanoma, glioma, and astrocytoma. TMZ exerts its effects mainly via the mutagenic product O(6)-methylguanine, a cytotoxic DNA lesion. This damage may be repaired by the DNA repair enzyme O(6)-methylguanine DNA methyltransferase (MGMT), a key player in the resistance of cancers to TMZ. Several strategies are presently being pursued to improve the killing of tumor cells by TMZ, with inhibition of MGMT being the most promising. In this review, we provide an overview of recent advances in this field.

Retrospective analysis of thyroid nodules by clinical and pathological characteristics, and ultrasonographically detected calcification correlated to thyroid carcinoma in South China.

OBJECTIVE: To investigate the clinical and pathological characteristics of thyroid nodules, as well as to evaluate the significance of ultrasonographically detected thyroid calcification in the diagnosis of thyroid carcinomas. METHODS: Retrospective data were studied from 1,051 consecutive patients who underwent a thyroidectomy in the Provincial Hospital of Fujian Medical University in South China between January 2003 and July 2006 for nodular thyroid disease. Complete sonographical information before surgery was only collected from 758 of the 1,051 patients. RESULTS: Among the 1,051 patients, benign lesions were found in 857 (81.54%) patients, of whom 612 (71.41%) were nodular goiter; malignant lesions were found in 194 (18.46%) patients, in whom benign thyroid lesions were also found in 85 (43.81%) patients. A total of 48 patients suffered from microcarcinomas, of whom 37 patients had benign lesions; these 37 accounted for 43.53 and 77.08%, respectively, of the 85 malignant cases with benign lesions and the 48 cases with microcarcinomas. In the 758 patients who underwent thyroid ultrasonography before surgery, intrathyroidal calcifications were apparent in 243 patients (32.06%). The incidence of calcification was significantly higher in patients with thyroid carcinoma (54.17%) than in those with benign lesions (26.87%; p < 0.005). Detection of calcification in thyroid lesions by ultrasound had a sensitivity of 32.38% and a specificity of 87.35%, with an OR of 3.31 (95% CI, 2.24-4.63), positive likelihood ratio of 2.56, negative likelihood ratio of 0.77 and a kappa value of 0.23. CONCLUSION: Thyroid carcinoma, especially microcarcinoma, often coexists with benign thyroid disease. Calcification detected by thyroid ultrasound represents a risk factor for malignancy, but is of limited use as a sole marker of malignancy.

Production of xylobiose from the autohydrolysis explosion liquor of corncob using Thermotoga maritima xylanase B (XynB) immobilized on nickel-chelated Eupergit C.

In this study, a thermostable recombinant xylanase B (XynB) from Thermotoga maritima MSB8 was immobilized on nickel-chelated Eupergit C 250L. This immobilized XynB was then used to hydrolyze the autohydrolysis explosion liquor of corncob (AELC) in a packed-bed enzyme reactor for continuous production of xylooligosaccharides, especially xylobiose. When tested in batch hydrolysis of AELC, the immobilized XynB still retained its relative activity of 92.5% after 10 cycles of hydrolysis at 90 degrees C. The immobilized XynB retained 83.6% of its initial hydrolysis activity even after 168 h of hydrolysis reaction at 90 degrees C and demonstrated a half-life time of 577.6 h (24 days) for continuous hydrolysis. HPLC showed that xylobiose (49.8%) and xylose (22.6%) were the main hydrolysis products yielded during continuous hydrolysis. Xylobiose was adsorbed on an activated charcoal column and eluted with a linear gradient of 15% (v/v) ethanol to yield xylobiose with 84.7% of recovery. Also, the purity of xylobiose was up to 97.2% as determined by HPLC. Therefore, the immobilized XynB was suitable for the efficient production of xylobiose from AELC. This is the first report on the immobilization of xylanase for xylobiose production.

Synergic CSE1L/CAS, TNFR-1, and p53 apoptotic pathways in combined interferon-gamma/adriamycin-induced apoptosis of Hep G2 hepatoma cells.

Many cancers are chemotherapy-resistant. Chemotherapy combined with immunotherapy offers a potential avenue for the treatment of chemotherapy-resistant cancers. In this study, we investigated the apoptotic pathways induced by combined interferon-gamma/adriamycin treatment in Hep G2 cells. Our data showed that Hep G2 cells treated with combined interferon-gamma/adriamycin enhanced cell apoptosis in comparison with that of cells treated with adriamycin. Interferon-y increased TNFR-1, CSE1L/CAS (cellular apoptosis susceptibility protein), Bax, and Bad levels. Adriamycin increased p53 and Bax, but not TNFR- 1 and CAS levels. Interferon-y did not increase p53 accumulation; nevertheless it enhanced adriamycin-induced p53 accumulation. Overexpression of IRF-1 augmented the combined interferon-gamma/adriamycin-induced p53 accumulation. Interferon-gamma co-treatment increased the stability of p53 protein induced by adriamycin. Our data suggest that TNF-gamma may greatly enhance the combined interferon-gamma/chemotherapeutic drug-induced apoptosis of cancers. Our findings also indicate that CAS, TN-FR-1, p53, Bax, and Bad may be the targets for the interferon-y-based chemo-immunotherapy of the chemotherapy-resistant cancers.

Effect of nicotinamide and pentoxifylline on normal tissue and FSA tumor oxygenation.

Nicotinamide (NA) and pentoxifylline (PTX) sensitize experimental murine tumors to radiation without sensitizing normal tissues. They are presumed to exert this effect by reducing hypoxia in tumors. The present study evaluated the individual and combined effects of NA and PTX on oxygen levels in subcutaneous normal tissue and subcutaneous FSa fibrosarcoma tumors in the hind foot dorsum of C3H mice. Oxygen measurements were made using a polarographic needle electrode inserted into the tissue immediately before and/or 15-60 min after intraperitoneal administration of 500 mg/kg of NA, 50 mg/kg of PTX, or saline. The median tumor pO2 increased from a mean +/- S.E.M. of 4.1 +/- 1.1 mm Hg in saline-treated control mice to 6.8 +/- 1.9 mm Hg 15 min after NA, 7.6 +/- 1.4 mm Hg 60 min after PTX, and 6.7 +/- 1.1 mm Hg after NA and PTX in combination. PTX raised the median tumor pO2 level from 21% to 39% of the median subcutaneous normal tissue pO2 (p < 0.01). PTX also significantly reduced the proportion of tumor pO2 values < or = 2 mm Hg from 41 +/- 10% to 8 +/- 7% (p = 0.02). Although NA did increase the proportion of tumor that was well oxygenated, it did not significantly reduce the proportion of tumor pO2 values < or = 2 mm Hg (p = 0.34). The combination of NA and PTX did not add to the tumor oxygenation enhancement achieved by PTX alone. NA increased the median subcutaneous normal tissue pO2 by an average of 5.1 +/- 2.2 mm Hg from a baseline of 17.1 +/- 2.2 mm Hg (p = 0.04). PTX had no effect on the median normal tissue pO2 (p = 0.93). PTX showed greater therapeutic potential in this model system than did NA.

The extent, time course, and fraction size dependence of mouse spinal cord recovery from radiation injury.

PURPOSE: This experiment was designed to assess: (a) the influence of fraction size and time interval between fractions on the tolerance of the spinal cord to high cumulative doses of radiation; and (b) the influence of the long-term recovery process on the tolerance of the spinal cord to reirradiation. METHODS AND MATERIALS: The T10-L2 level of the spinal cord of C3Hf mice was irradiated using a conventionally fractionated regimen of 2.0 Gy once daily, a prolonged fractionated regimen of 1.2 Gy once daily, a hyperfractionated regimen of 1.2 Gy twice daily, or a single dose of 12 Gy followed 0-190 days later by a second dose of 5-20 Gy. Mice in the multifractionated regimen groups were given a single 15 Gy top-up dose 24 h after reaching a cumulative fractionated dose of 24-70 Gy. Hind limb strength was measured weekly for 2 years after the completion of irradiation. RESULTS: Paralysis occurred in a bimodal time distribution, with peaks at 5-10 months and 15-23 months after the completion of irradiation. The cumulative radiation dose was directly associated with the incidence of paralysis in each radiation schedule (p < 0.0001) and inversely associated with the time to onset of paralysis in the 1.2 Gy b.i.d. (p = 0.0001) and 2.0 Gy q.d. schedules (p = 0.03). The median latency of paralysis in each group was inversely associated with the incidence of paralysis in that group (p < 0.001). Decreasing the fraction size from 2.0 to 1.2 Gy once daily markedly increased the radiation tolerance of the spinal cord (p < 0.0001), consistent with a very small alpha-beta value of -0.30 Gy (approximately 95% confidence interval -0.72, +0.18) in the linear-quadratic model. Decreasing the time interval from 24 h to alternating 8 and 16 h periods produced an offsetting diminuation in cord tolerance (p < 0.0001). The 1.2 Gy once daily schedule resulted in ED20 and ED50 values that were approximately double those of the 2.0 Gy once daily and the 1.2 Gy twice daily schedules and a relative risk of paralysis from a given dose that was 0.03 times the risk associated with the other two regimens (p < 0.0001). There was no significant difference between the 2.0 Gy once daily and the 1.2 Gy twice daily dose-paralysis curves (p = 0.86). The residual from a single 12 Gy radiation dose was 17% after 190 days, leaving the retreatment ED50 only 10% below the ED50 of previously unirradiated spinal cord. The relative risk of paralysis after 12 Gy plus a second radiation dose decreased from 1.00 with no time interval between doses to 0.51-0.73 with a 0.25, 1 or 3 day interval, 0.32 with a 7 day interval, 0.11 with a 30 day interval, and 0.06 with a 190 day interval. CONCLUSION: The increased radiation tolerance of the murine spinal cord produced by decreasing the fraction size from 2.0 to 1.2 Gy was offset by the diminished tolerance produced by decreasing the time interval between fractions from 24 to 8-16 h, resulting in no significant difference in the dose-paralysis curves of conventional and hyperfractionated radiation schedules. The rodent spinal cord eliminates the majority of the occult radiation injury produced by a radiation dose equal to half the ED50 during the months following irradiation. This permits retreatment of previously irradiated spinal cord to high doses without the induction of myelopathy.

Human melanoma antigen AH is an autoantigenic ganglioside related to GD2.

AH antigen, initially defined by an antibody present in a melanoma patient, is a cell surface antigen found on approximately 65% of melanoma cell lines. Absorption analysis indicates that AH is a differentiation antigen marking normal and malignant cells of neuroectodermal origin. The AH determinant has been found to be related to GD2 ganglioside.

Cell surface antigens of human malignant melanoma: definition of six antigenic systems with mouse monoclonal antibodies.

Eighteen mouse monoclonal antibodies were selected for reactivity with cell surface antigens of the immunizing human melanoma cell line SK-MEL-28. Six distinct antigenic systems were defined by direct serological assays and absorption tests with a panel of 41 cell lines derived from normal and malignant human tissues. Biochemical analysis indicated that two of the antigens are glycoproteins with molecular sizes of 95,000 and 150,000 daltons (gp95 and gp150). Two other antigenic systems (O5 and the R24 group) are associated with heat-stable molecules having the characteristics of glycolipids. The remaining two antigens (M19 and R8) are heat labile, but molecular characterization has not been possible. Each of the antigenic systems has a distinctive pattern of distribution on various cell types, varying from a broad representation to a more restricted occurrence. O5 appears to be a species antigen, being present on virtually every human cell type tested. gp95, gp150, M19, and R8 are found on a characteristic proportion of melanomas, astrocytomas, and epithelial cancers and on normal kidney cells. The antigen defined by the R24 antibody has the most restricted distribution of all. Reactivity is found with melanomas and astrocytomas, whereas epithelial cell types, fibroblasts, and cells of hematopoietic origin lack R24. Although occurrence of gp95, gp150, M19, and R8 distinguishes a small subset of melanomas not expressing these antigens, R24 is found on all melanoma cells.

Cell surface antigens of human renal cancer defined by autologous typing.

Sera from 28 patients with renal cancer were tested for reactivity with surface antigens of cultured autologous renal cancer cells. Four serological assays were used to survey sera for autologous antibody. Immune adherence, protein A, and C3-mixed hemadsorption assays detected reactivity in a high percentage of patients (80-100%), whereas mixed hemadsorption assays were negative with sera from all but one patient. Reactive sera from six patients were analyzed by absorption tests with autologous, allogeneic, and restricted to autologous renal cancer cells; class 2 antigens, present on certain allogeneic renal and nonrenal cancer cells; and class 3 antigens, found on a wide variety of normal and malignant cell types. The sera of one patient detected class 1, 2, and 3 antigens, the sera of three patients detected class 2 antigens, and the sera of two patients detected class 3 antigens. This analysis of renal cancer, with the recognition of three classes of surface antigens recognized by autologous sera, resembles the results of autologous typing of three other human malignancies: malignant melanoma, acute leukemia, and astrocytoma. Evidence provided by autologous typing of these cancers indicates that class 1 and class 2 antigens are tumor-restricted and that under certain circumstances these antigens are immunogenic for the autologous host.