Abstract:Objective To study the effect of tubulin tyrosine ligase like 12 (TTLL12) and nitrotyrosine on the growth of prostate cancer cells. MethodsTTLL12 and α-tubulin nitrotyrosination (N-tubulin) were detected by Western blotting analysis in normal prostate epithelial cell lines PWR1E and RWPE1, and prostate cancer cell line DU145; the proliferation of cells was observed by MTT assay. ResultsThe expression of N-tubulin was significantly lower in DU145 cells than those in PWR1E and RWPE1 cells after treatment with nitrotyrosine (P＜0.05). The proliferation of PWR1E and RWPE1 cells treated with nitrotyrosine (cultured with 800 μmol/L nitrotyrosine) were significantly inhibited compared with the control (cultured with 800 μmol/L hydrochloric acid, P<0.05), while there was no visible change in the proliferation of DU145 cells. N-tubulin level was significantly higher in the experimental group (transfected with siTTLL-12) than the control group (transfected with siControl) after effective silence of TTLL12 (P＜0.05); the proliferation of DU145 cells treated with nitrotyrosine in the experimental group was significantly inhibited (P<0.05),while there was no visible change in cell proliferation in the control group (P>0.05). ConclusionTTLL12 allows an escape of prostate cancer cells from the treatment of nitrotyrosine,leading to the escape of prostate cancer cells from the body’s surveillance system and acquirement of their abnormal proliferation.

Abstract:Objective To identify the sex-related DNA copy number alterations(CNA) in hepatocellular carcinoma(HCC). MethodsHigh-resolution array comparative genomic hybridization(array-CGH) was used to examine 17 female and 46 male HCCs. Two-tailed Fisher’s exact test or χ2 test was used to compare the differences in CNA between females and males. ResultsThe overall frequencies and patterns of CNA in female and male cases were similar. However, female HCC tumors presented more copy number gains compared to male on 1q21.3-q22(76.5% vs 37.0%, P=0.009), 11q11(35.3% vs 0.0%, P=0.000 2) and 19q13.31-q13.32(23.5% vs 0.0%, P=0.004), and more loss on 16p11.2(35.3% vs 6.5%, P=0.009). Relative to females, male cases had more copy number loss on 11q11(63.0% vs 17.6%, P=0.002). Further analyses showed that 11q11 gain was correlated with 19q13.31-q13.32 gain(P=0.042) and 16p11.2 loss(P=0.033), while 1q21.3-q22 gain was correlated with 19q13.31-q13.32 gain(P=0.046). ConclusionOur findings suggest that CNA may play an important role in sex-related difference in HCC development.

Abstract:Objective To create transient receptor potential vanilloid 6 (Trpv6) gene knockout mouse model, so as to pave a way for further research of its biological function and its role in bone metabolism in vivo. MethodsMouse genomic DNA sequence of Trpv6 gene was obtained from Ensembl database. Trpv6 gene knockout vector (pBR322-MK-Trpv6) was constructed. Trpv6 knockout vector was transferred into the embryonic stem (ES) cells by electroporation and screening of both G418 and Ganciclovoir resistant clones were performed routinely. The homologous recombined ES cell clones were identified by PCR. The correct homologously recombined ES cells were microinjected into C57BL/6J mouse blastocysts to obtain chimera mouse. Male mice with a chimera rate of 50% were mated with C57BL/6J female mice; the offsprings with gray fur were obtained, which were identified as heterozygote mice by PCR. Heterozygote mice were intercrossed to generate homozygote mice. ResultsTargeting vector pBR322-MK-Trpv6 were successfully constructed. A total of 24 correct homologously recombined clones were gained after electroporation. The efficiency of homologous recombination was 25%. Four male mice with a chimera rate of more than 50% were acquired after homologously recombined clones through microinjection. After the chimera mice were mated with C57BL/6J mice, 57 grey-fur mice originated from ES cell were gained, including 17 (29.8%) with heterozygous genotype. Heterozygote mice were intercrossed to generate homozygote mice. Western blotting analysis showed no Trpv6 protein expression in homozygote mice. ConclusionWe have successfully established Trpv6 gene knockout mouse model, and there is no embryonic lethality in homologous mutant mice.

Abstract:Objective To investigate a new method for creating experimental model of noninvasive fungal sinusitis in rabbits. MethodsTwenty-five New Zealand white rabbits(50 sides)were divided into seven groups: control group (n=6 sides),sham-operation group (n=6 sides),mucosal-scratched group (n=6 sides),ostia-blocked group(n=6 sides), Aspergillus fumigatus inoculated group(n=8 sides), mucosal-scratched plus Aspergillus fumigatus inoculated group(n=9 sides),and ostia-blocked plus Aspergillus fumigatus inoculated group(n=9 sides).The experiment was divided according to whether the maxillary sinus mucosa was wounded or not and whether the natural ostia were blocked or not. On days 14 and 28 after Aspergillus fumigatus inoculation, histopathologic and mycological examinations of both maxillary cavities and mucous membrane were performed to confirm the establishment of the model. ResultsAspergillus fumigatus plus ostial obstruction group had a positive rate of 88.9% (8/9 sides) on day 14 and 77.8% (7/9 sides) on day 28 in fungal culture. Aspergillus fumigatus plus mucosa wound yielded a positive rate of 44.4%(4/9 sides) on day 14 and 22.2%(2/9 sides) on day 28. ConclusionAspergillus fumigatus inoculation combined with an blocked ostium can be used to establish experimental model of noninvasive maxillary fungal sinusitis.

Abstract:Objective To study the role and related mechanisms of pax5 gene during the development of the embryos, especially in the early development of B cell and midbrain, and to observe the expression of pax5 gene in wildtype zebrafish embryos. MethodsTotal RNA of Tuebingen wildtype zebrafish embryos was extracted to obtain cDNA of pax5 gene by RT-PCR with specific primers. The cDNA of pax5 gene and pCS2+ vector were double digested with EcoRⅠ and XbaⅠ, and then ligated by T4 DNA ligase. The recombinant vector was verified by double digestion, colony PCR screening and sequencing. The verified recombinant vector was then used to synthesize digoxin labeled anti-sense mRNA probe of pax5 gene using T3 RNA polymerase in vitro transcription system. The generated probes were used to detect pax5 gene expression in zebrafish embryos by whole-mount in situ hybridization. ResultsThe pCS2+-pax5 recombinant plasmid and the probe of digoxin-labeled anti-sense mRNA of pax5 gene was successfully constructed. Whole-mount in situ hybridization showed that pax5 gene was expressed in the cerebellum and midbrain hindbrain boundary from 18-72 h post-fertilization(hpf). In the cochlea pax5 gene was expressed from 24-72 hpf and the expression was increased as time went by. From 18-48 hpf pax5 gene is found in the notochord. ConclusionPax5 gene is highly expressed in the brain and notochord of zebrafish embryos. Pax5 gene expression is found in the cochlea of the Tuebingen wildtype zebrafish’s embryos for the first time. It is suggested that pax5 may play an important role in the development of the nervous system in the early zebrafish embryos.

Abstract:Objective To investigate the effect of liver X receptor α (LXRα) gene silencing on lipid metabolism-related genes in HepG2.2.15 cells. MethodsHepG2.2.15 cells were divided into blank control group (without transfection), negative control group (transfected with HK plasmid), and shLXRα group(transfected with shLXRα plasmid). The shLXRα plasmids carrying LXRα gene were constructed and were used to transfect HepG2.2.15 cells using PolyJetTM reagent. Green fluorescent protein and LXRα protein expression were examined by fluorescence microscope and Western blotting analysis 24-96 h after transfection, so as to identify the best interference time. Then cells were treated with agonist T0901317 for 24 h or 48 h; and the content of triglyceride (TG) was observed to detect the degree of steatosis by biochemical assay. The expression of sterol regulatory element binding protein-1c (SREBP-1c) mRNA was detected by RT-PCR and the expression of hepatitis B virus X (HBx) protein and fatty acid synthase (FAS) protein was tested by Western blotting analysis. ResultsThe shLXRα plasmid was constructed and transfected into HepG2.2.15 cells successfully. Compared with blank and negative control groups, LXRα protein was markedly decreased in the shLXRα group, with the lowest level found at 48-72 h after transfection (P＜0.01). After cells were stimulated with T0901317, HBx and FAS protein expression, the content of TG, and SREBP-1c mRNA expression gradually increased with the prolongation of stimulation period, and there was no significant difference in HBx expression at the same time point between different groups. FAS protein, TG contents, and SREBP-1c mRNA in shLXRα group were significantly lower than those in the other two groups (P<0.01). ConclusionHBx can regulate lipid metabolism through LXRα/SREBP-1c/FAS pathway.

Abstract:Objective To investigate the expression of Toll-like receptor 4 (TLR4), NF-κB p65 and the negative regulating factor Toll-interacting protein (Tollip) in colonic mucosa of rats with 2，4，6-trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis. MethodsForty healthy male SD rats were randomly divided into normal control group and model group. The expression of TLR4, NF-κBp65 and Tollip was detected and compared between the two groups by immunohistochemistry(IHC) and RT-PCR. ResultsRT-PCR revealed a significant higher expression of TLR4 and NF-κB p65 mRNA in the model group compared with the normal control group (TLR4: 0.376±0.029 vs 0.215±0.049, t= 2.731, P=0.013； NF-κB p65: 0.746±0.049 vs 0.206±0.063, t=6.055, P=0.000). The expression of TLR4 and NF-κB p65 was positively correlated with the general injury score (TLR4: r=0.754, P=0.000; NF-κB p65: r=0.548, P=0.012) and the histological injury score (TLR4: r=0.866, P=0.000; NF-κB p65: r=0.919, P=0.000). Expression of Tollip mRNA was not significantly different between the two groups as detected by RT-PCR (0.288±0.050 vs 0.140±0.046, t=1.993, P=0.061). IHC revealed that Tollip protein expression was mainly assembled in the colonic mucosal lamina propria, and the expression in the model group was higher than that in the normal control group. Tollip expression was negatively correlated with the general injury score (r=-0.497, P=0.026) and the histological injury score (r=-0.551，P=0.012). ConclusionThe unbalanced expression of TLR4 and the negative regulating factor Tollip participates in the pathogenesis of inflammatory bowel disease.

Abstract:Objective To observe the expression of YKL-40 protein in the pulmonary tissues of rats with bleomycin-induced pulmonary fibrosis(PF). MethodsThe PF model group was induced with intratracheal instillation of bleomycin solution (7.5 mg/kg) and the control group was treated with normal saline. On day 7, 14, 21 after bleomycin challenge, rats were sacrificed and the pulmonary tissues were harvested. H-E staining, Masson staining and Szapiel score were employed to determine alveolitis and pulmonary fibrosis. Expressions of YKL-40 in lung tissues were detected by real-time RT-PCR, Western blotting analysis and immunohistochemistry method. ResultsBleomycin instillation induced alveolitis in the lung of rats, with inflammation score being significantly higher on day 7 (2.8±0.45, P<0.01) and on day 21 (1.8±0.84, P<0.05) compared with that of control group (0.42±0.25). Pulmonary fibrosis degrees in model group was significantly higher on day 14 (1.7±0.73, P<0.05) and on day 21 (2.9±0.56, P<0.01) compared with that of control group (0.2±0.45). YKL-40 mRNA (YKL-40/β-actin) expression was significantly increased on day 7 (3.71±0.25) after bleomycin treatment compared with the control group (P<0.05). Western blotting analysis showed that YKL-40 protein expression was significantly increased on day 14(0.56±0.24,P<0.05) and on day 21(1.15±0.19, P<0.01) after bleomycin treatment compared with the control group (0.23±0.07). The results of immunohistochemistry showed that bleomycin up-regulated YKL-40 expression in a time-dependent manner, and YKL-40 expression was mainly located in the smooth muscle cells, alveolar macrophages, alveolar epithelium and fibroblasts. ConclusionYKL-40 expression may contribute to the pulmonary fibrosis, and may participate in the pathogenesis of pulmonary fibrosis.

Abstract:Objective To discuss the relationship of CXCR4 expression with clinical pathology, post-operative metastasis, recurrence and prognosis of gastric cancer patients. MethodsA total of 141 gastric cancer tissues and the corresponding adjacent tissues were taken from patients who were treated in the Department of General Surgery, The Affiliated Provincial Hospital of Anhui Medical University between November 2007 to November 2008.Immunohistochemistry was performed to detect the CXCR4 expression in the gastric cancer tissues and adjacent tissues; the relationship of CXCR4 expression with clinical pathology of the patients was analyzed. The relationship of CXCR4 expression with recurrence and prognosis was analyzed based on a 3 year-follow-up. ResultsThe positive rate of CXCR4 was 60.3%(85/141) in gastric cancer tissues, which was significantly higher than that in the corresponding adjacent tissues 26.2%(37/141, P<0.05). CXCR4 expression in the gastric cancer tissues was positively correlated with the TNM stage, depth of invasion, invasion of the pancreas capsule, and lymphatic metastasis (P<0.05).Logistic multivariate regression analysis showed that lymphatic metastasis and recurrence after surgery were correlated with the expression of CXCR4 in gastric cancer tissues(P<0.05).The 3-year survival rate of patients with CXCR4 expression was significantly lower than patients without CXCR4 expression(43.1% vs 56.6%,P<0.05). ConclusionCXCR4 is over-expressed in gastric cancer tissues, and the expression is positively correlated with the lymphatic metastasis and recurrence after surgery. CXCR4 positive patients have a lower survival rate than patients without CXCR4 expression; all these makes CXCR4 a possible molecular marker for lymphatic metastasis, postoperative recurrence and prognosis in gastric cancer.

Abstract:Objective To observe the effects of voluntary exercise on the learning ability, memory and hippocampus growth-associated protein 43 (GAP43) expression in senescence-accelerated prone mouse (SAMP8), so as to explore the possible mechanism of exercises on improving the cognitive ability and delaying aging. MethodsA total of 60 three-month old female SAMP8 mice were evenly assigned to running cage environment (RCE) group and standard environment (SE) group at random. After three months, Morris water maze test was used to test the platform-seeking latency and search strategy. Then 10 mice were sacrificed in each group for RT-PCR analysis of hippocampus GAP43 mRNA expression, 10 for Western blotting analysis of hippocampus GAP43 protein expression, and 10 for immunohistochemistry staining of hippocampus GAP43 expression. ResultsMorris water maze test showed that RCE mice had a significant shorter platform-seeking latency than SE mice(P<0.01, P<0.05), and RCE mice had a significant longer time in the first quadrant (P<0.01) and a shorter time in the fourth quadrant (P<0.05) compared with SE mice. RCE mice had a significantly higher GAP43 expression in the hippocampus compared with SE mice (P<0.01). ConclusionVoluntary exercise can improve the learning ability and memory of SAMP8, which might be associated with the increase of GAP43 in the hippocampus.

Abstract:Objective To explore the factors affecting the circadian rhythm of blood pressure in treated hypertensive patients. Methods Based on the clinical data, ambulatory blood pressure, 24-hour mean blood pressure (24hMBP) and circadian rhythm, 125 consecutive patients receiving antihypertensive treatment were divided into the following groups: normal 24hMBP and dipper type (typeⅠ), abnormal 24hMBP and dipper type(typeⅡ), abnormal 24hMBP and non-dipper type(typeⅢ), and normal 24hMBP and non-dipper type (type Ⅳ). The clinical data and ambulatory blood pressure of the latter three groups were compared. Non-conditional Logistic regression was used to analyze the factors for ambulatory blood pressure. Results Ninety patients had abnormal 24hMBP (72.0%), and 106 (84.8%) had abnormal circadian rhythm. Among these cases, 2 (1.6%) patients had typeⅠ, 17 (13.6%) had typeⅡ, 73 (58.4%) had type Ⅲ, and 33 (26.4%) had type Ⅳ. The incidence of kidney disease in patients with abnormal 24hMBP was significantly higher than those with normal 24hMBP (P<0.05). The incidence of diabetes mellitus in patients with abnormal circadian rhythm (non-dipper type) was higher than those with normal circadian rhythm (dipper type), with the highest incidence seen in patients of type Ⅳ (P<0.05); the rates of calcium channel blocker (CCB) use and a daily morning dose administration in type Ⅳ patients were higher than those in type Ⅱ and type Ⅲ patients (P<0.05). Type Ⅳ group had the most severe abnormal ambulatory blood pressure among the latter three groups. Logistic regression analysis showed that complication with renal diseases (OR=0.301,95%CI:0.124~0.729, P=0.008), use of CCB (OR=2.191,95%CI: 0.967~4.966,P=0.048), and administration of a morning dose (OR=2.384,95%CI: 1.017~5.591,P=0.046) were the factors of ambulatory blood pressure. Conclusion Abnormal circadian rhythm of blood pressure is high in patients receiving antihypertensive treatment. Complication with kidney diseases, use of CCB, and a daily morning dose are the factors for ambulatory blood pressure, indicating it is be reasonable to analyze factors for circadian rhythm by combining the 24hMBP and the components of circadian rhythm.

Abstract:Objective To study the values of standard 12-lead electrocardiogram (ECG) in differential diagnosis of acute pulmonary embolism (APE) and non-ST elevation myocardial infarction (NSTEMI). MethodsA retrospective analysis was conducted on 126 patients who were treated in the First Affiliated Hospital of Wenzhou Medical College during Jan. 2005 to Jan. 2011. The patients included 42 patients with APE (mean age [61±12] year) and 84 with NSTEMI (mean age [72±15] year). The data of all patients were complete and true, and the two groups were comparable in age and sex distribution. The standard 12-lead ECG records on admission were analyzed and the parameters which could be used for differential diagnosis were screened. Results Frequencies of right bundle branch block (RBBB) and SⅠQⅢTⅢ or SⅠSⅡSⅢ pattern were similar in the two groups ([11.9%] APE vs [14.3%] NSTEMI, [26.2%] APE vs [15.5%] NSTEMI patients, respectively). Negative T waves in leads Ⅴ1-Ⅴ3 together with negative T waves in inferior wall leads Ⅱ, Ⅲ, aVF (OR 1.32, 95%CI[1.15-1.69]) yielded a positive predictive value of 88% and specificity of 82% for APE. However, ST depression in leads Ⅴ5-Ⅴ6 and negative T waves in leads Ⅴ5-Ⅴ6 (OR 1.85, 95%CI [1.14-3.01]) yielded a positive predictive value of 89% and specificity of 50% for NSTEMI. ConclusionRBBB and SⅠQⅢTⅢ or SⅠSⅡSⅢ pattern may not help to differentiate between APE and NSTEMI. Coexistence of negative T waves in precordial leads Ⅴ1-Ⅴ3 and inferior wall leads Ⅱ, Ⅲ, aVF may suggest APE diagnosis. Coexistence of negative T waves and ST segment depression in precordial leads Ⅴ5-Ⅴ6 may suggest NSTEMI diagnosis.

Abstract:Objective To investigate the expression of TNF receptor-associated factor 6 (TRAF6) in the peripheral blood mononuclear cells (PBMCs) of primary Sjgren’s syndrome (pSS) patients and its clinical significance. Methods The expression of TRAF6 mRNA was determined by quantified reverse transcriptional PCR in the PBMCs of 23 pSS patients and 23 healthy controls. The correlation of TRAF6 mRNA level with the laboratory findings, including serum rheumatoid factor (RF), globulin, anti-SSA and anti-SSA antibody, were analyzed by Spearman test. Nine of the pSS patients were followed up and their TRAF6 mRNA expression was detected after glucocorticoid treatment for 1-2 months. Results The TRAF6 mRNA level in pSS patients was 2.77 times higher than that in the healthy controls(P<0.01). The TRAF6 mRNA level in pSS patients was positively correlated with serum RF (r=0.45, P=0.03) and globulin (r=0.43, P=0.04), but was not associated the anti-SSB and anti-SSA antibody. TRAF6 mRNA level was decreased by about 50% in the patients who were treated with glucocorticoids for 1-2 months (P<0.05). Conclusion TRAF6 in PBMCs is involved in the pathogenesis of pSS and it may be a potential biomarker for pSS.

Abstract:Objective To characterize the serum metabolite composition of patients with clear cell renal cell carcinoma (ccRCC) by using nuclear magnetic resonance 1H (1HNMR) spectroscopy. Methods The 1HNMR spectra were obtained for the serum samples of 11 patients with low grade（Ⅰ-Ⅱ） ccRCC and 16 healthy humans, and the spectra data were analyzed by principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) to demonstrate the metabonomic changes. Results PCA and PLS-DA results showed that there were noticeable differences in metabonomics between the ccRCC patients and healthy controls. The levels of low-density lipoprotein, very low-density lipoprotein, isoleucine, alanine, N-acetylglycoprotein, acetoacetate, glutamine, taurine, 1-methylhistidine, and phenylalanine were significantly increased in the ccRCC patients (P<0.05, P<0.01); and lactate, acetate, pyruvate, choline, glycerol, and glucose were significantly decreased compared with those in the healthy controls (P<0.05, P<0.01). Conclusion Serum 1HNMR spectroscopy combined with PCA and PLS-DA can identify the difference in serum metabonomes between ccRCC patients and healthy controls, which may offer a novel way for diagnosis of ccRCC.

Abstract:Objective To prepare an absorbable crosslinked microporous hemostatic starch(ACMHS), to observe its hemostasis effect in the liver, kidney and femoral artery, and to discuss the hemostasis mechanism. Methods Thirty adult healthy New Zealand rabbits of either sex, weighing 2,000-2,250 g, were equally randomized into 3 groups: liver bleeding group (group A), kidney bleeding group (group B) and femoral artery bleeding group (group C). Each group was further divided into a test group and a control group randomly (n=5) according to different hemostatic powders; ACMHS was used in the test group(group A1, B1, C1), and AristaTM was used in the control group (group A2, B2, C2). The bleeding amount and bleeding time were accurately recorded. The pathological changes at incision and in the surrounding tissues of the liver, kidney and femoral artery were observed by hematoxylin-eosin staining after test. Results All the bleeding amounts of group A1 and A2 were (1.02±0.10) g and (1.03±0.09) g, and the bleeding time periods of group A1 and A2 were (92.6±5.16) s and (95.6±5.31) s, respectively (P>0.05). All the bleeding amounts of group B1 and B2 were (0.92±0.09) g and (0.94±0.08) g, and the bleeding time periods of group B1 and B2 were (87.5±7.48) s and (88.8±6.54) s, respectively (P>0.05).All the hemostatic efforts failed in group C. Hematoxylin-eosin staining showed accumulation of red blood cells at the incision and in surrounding tissues of the liver and kidney in group A and group B, without evidence of burn. Conclusion ACMHS has the same hemostatic function as AristaTM in parenchymatous organ injury, but it is not suitable for bleeding of femoral artery injury with high pressure.

Abstract:Objective To investigate the expression of macrophage migration inhibitory factor (MIF) in complex regional pain syndrome Ⅰ (CRPS Ⅰ) rat model and the possible efficacy of MIF blockage in treatment of CRPS Ⅰ. Methods Fifty healthy male SD rats were randomly divided into the following 5 groups: control, sham, model, DMSO control, and ISO-1(inhibitor of MIF) treatment. CRPS1 models were created in the last 3 groups; rats in the ISO-1 treatment group were subcutaneously treated with ISO-1 dissolved in 10 μl 5% DMSO at 1 mg/(kg·d) for 14 d. DMSO control group was only given 10 μl 5% DMSO. The pain threshold and thickness of the hindpaws were measured before and after treatment and were compared. The levels of MIF protein were examined in the serum, skin, spinal cord, sciatic nerve, and cerebrospinal fluid using ELISA and Western blotting analysis.Results Rats in the model group had noticeable hindpaw edema and significantly decreased pain threshold compared with the baselihe and that of the control group(P<0.01) . The hindpaw edema and pain threshold were significantly improved in ISO-1 treatment group compared with those in the model group(P<0.05). MIF levels in the serum, skin, spinal cord, cerebrospinal fluid and sciatic nerve were higher in the model, DMSO control, and ISO-1 groups compared with those in the control group (P<0.05 or P<0.01). Conclusion MIF is a key inflammatory factor of CRPS Ⅰ, and anti-MIF treatment might be a new therapy for human CRPS Ⅰ.

Abstract:Objective To study the value of magnetic resonance spectroscopic imaging (MRSI) with only surface coils in diagnosing prostate cancer. Methods A total of 48 elderly patients with surgery or biopsy-confirmed benign prostatic hyperplasia or prostate cancer were preoperatively examined by surface coils for three-dimensional simple multi-voxel magnetic resonance spectroscopy scans, and the voxel (choline[Cho]+creatine[Cr])/citrate(Cit) values were calculated. According to the pathological findings, the voxels were divided into prostate cancer and non-cancer. The voxel (Cho+Cr)/Cit values of prostate central zone and prostate peripheral zone were analyzed. The voxel nature of (Cho+Cr)/Cit value was used as the gold standard receiver to plot characteristic curve (ROC curve) and to calculate the area under the curve; the maximal Youden index was used as the standard to obtain the cut-off value and the corresponding sensitivity and specificity. Results The voxel (Cho+Cr)/Cit values of different natures were significantly different (P<0.001). When the area under the ROC curve of (Cho+Cr)/Cit value was used for diagnosis of prostate cancer, the central zone was 0.652, and the peripheral zone was 0.821. For the cut-off value, the central zone was (Cho+Cr)/Cit=0.645, with the corresponding sensitivity and specificity being 0.512 and 0.71, respectively; the peripheral zone was (Cho+Cr)/Cit=0.815, with the corresponding sensitivity and specificity being 0.72 and 0.84, respectively. Conclusion Surface coil alone can successfully complete the three-dimensional proton spectroscopic imaging of prostate, and (Cho+Cr)/Cit=0.815 for the cut-off value has a high diagnostic value for peripheral zone prostate cancer.

Abstract:Objective To investigate the effectiveness of systemic moist therapy in treating stage Ⅱ-Ⅳ pressure ulcers. Methods From January 2010 to January 2011, 21 males and 9 females with pressure ulcers,with an age range of 60-80 years and a mean of 71±8.5 years old, were treated with systemic moist dressing changes. Seven cases had Ⅱ degree pressure ulcers, 12 had Ⅲ degree and 11 had Ⅳ degree. Wound granulation and wound healing were observed after systemic moist treatment. Results The wounds were full of flesh granulation and the infection was controlled after the moist dressing therapy; all the wounds were healed within 1-6 weeks. A followed up of 3 months found no recurrence. Conclusion Moist therapy is a satisfactory method to treat deep pressure ulcers, and it is worth popularizing.

Abstract:Oxidative stress is implicated in the pathogenesis and progression of various diseases as well as in the natural aging process. DJ-1/PARK7 gene encodes DJ-1 protein, which participates in malignant transformation, sperm maturation and fertilization, cell proliferation promotion, and anti-oxidative stress. The anti-oxidative stress effect of DJ-1 has been drawing increasing attention in recent years. There have been related in vitro, in vivo and clinical studies. The main mechanisms of its anti-oxidation effects included modifying configuration, playing the role of molecular chaperones, protecting the mitochondrial function, and regulating anti-apoptosis genes and antioxidant-related genes. Further research on DJ-1 may provide a new strategy for the prevention and treatment of oxidative stress-related diseases.

Abstract:Apolipoprotein A5 (ApoA5) is a newly discovered apolipoprotein which is closely associated with high-density lipoprotein (HDL) remodeling. Clinically it demonstrates potent triglyceride reducing effect. Recent studies have found that ApoA5 can increase cholesteroal reverse transportation and HDL function by remodeling HDL, and it can also influence the progression of atherosclerosis in the different ways, showing a potential anti-atherosclerosis effect. Here we briefly review the characteristics of ApoA5 and its potential protective effect against atherosclerosis.

Abstract:Polylactide-co-glycolic acid (PLGA) tissue engineering scaffolds play an important role in the regeneration of bone and cartilage. Due to the poor hydrophilicity of PLGA, it is difficult for cells to attach to the scaffolds. Modification by RGD (Arg-Gly-Asp) peptides can effectively improve the cellular affinity of PLGA and adhesion and proliferation of the seed cells. This review summarizes the recent progress in PLGA tissue engineering scaffolds modified by RGD peptides.

Abstract:Objective To study the relationship of different ages with harvesting of ligamentum flavum and cell growth rate in primary culture, so as to improve the success rate of human ligamentum flavum cells(LFCs) culture in vitro. Methods A total of 21 surgical ligamentum flavum specimens from patients with intervertebral disc herniation and lumbar spinal stenosis were primary cultured with tissue block enzymolytic method. Cell outgrowth, morphology and growth status were observed under an inverted phase microscope. Expression of vimentin and type Ⅰ collagen was detected in the cells of third passage by immunofluorescence staining. The correlation between age and cell growth rate was analyzed. Results Cell morphology and growth status were well under inverted phase microscope and immunofluorescence staining showed vimentin and collagen type Ⅰ expression in the all LFCs. The cell growth rate of primary culture was negatively correlated with the age of patients from whom ligamentum flavum had been collected(r=-0.618, P<0.05). Cell growth rate was high when the patients were young. Conclusion Ligamentun flavum specimens should be taken from young subjects to ensure the success rate of primary culture of LFCs.

Abstract:Objective To investigate the serum level of high mobility group box-1(HMGB-1) in patients with liver cancer and its correlation with p53 and clinical pathological features of liver cancer. Methods Serum levels of HMGB-1 and p53 were determined by ELISA in 20 patients with hepatocellular carcinoma, 20 with cholangiocellular carcinoma and 20 healthy volunteers. Results Serum HMGB-1 levels in hepatocellular carcinoma patients ([83.8±17.4] ng/ml) and cholangiocellular carcinoma patients ([95.9±21.4] ng/ml) were significantly higher than that in the healthy volunteers ([8.4±2.6] ng/ml, P<0.05). Serum p53 levels in hepatocellular carcinoma patients ([251.0±45.6] pg/ml) and cholangiocellular carcinoma patients ([267.8±47.4] pg/ml) were also significantly higher than that in the healthy volunteers ([160.0±28.4] pg/ml, P<0.05). Serum HMGB-1 level was positively correlated with the serum p53 level in the hepatocellular carcinoma patients (r=0.660 3, P<0.05) and cholangiocellular carcinoma patients (r=0.614 2, P<0.05). Serum HMGB-1 and p53 levels were not correlated with the age, gender, tumor size, or number of tumors (P>0.05), but were correlated with the TNM stage, differentiation degree and metastasis status (P<0.05).Conclusion Serum HMGB-1 level is positively correlated with p53, and both HMGB-1 and P53 influence the pathogenesis of liver cancer.

Abstract:Objective To analyze the relationship of CAS protein expression with proliferative index, apoptosis index and clinical parameters in breast cancer tissues. Methods Immunohistochemistry for CAS expression, ki-67 (proliferative index) and TUNEL (apoptosis index) were examined in 20 usual ductal hyperplasia (UDH) , 20 atypical ductal hyperplasia (ADH), 10 ductal carcinoma in situ (DCIS), 53 invasive ductal carcinoma(IDC) and 14 normal breast tissues. Results CAS expression increased in order in the normal breast tissues, UDH, ADH, DCIS and IDC, with the positive rates of CAS protein being 14.3% (2/14), 25.0% (0/20), 40.0% (8/20), 60.0% (6/10), and 75.5% (40/53), respectively (χ2=29.382, P=0.000). CAS protein expression was correlated with histological grade, mitotic activity, and lymph node metastasis of IDC (P<0.01, P<0.05), and not with patient age, tumor volume or grade. CAS protein expression was positively correlated with ki-67 index (r=0.439, P=0.003), and not with the apoptosis index (r=0.248, P=0.083). Conclusion CAS protein expression is associated with cell proliferation index in breast cancer tissues.

Abstract:Objective To evaluate the pharmacokinetics and the relative bioavailability of pyridostigmine bromide sustained-release tablets (PBSTs) by comparing with the conventional tablets using a multiple-dose design. Methods Six rabbits were randomly divided into 2 groups and were assigned to a self crossover design. The plasma pyridostigmine bromide concentration was determined by high-performance liquid chromatography as multiple oral dose of PBSTs (90 mg each time, and twice a day) or conventional tablets (60 mg each time, three times a day). The pharmacokinetic parameters and relative bioavailability were calculated. Results A two-compartment model was used to describe the in vivo behavior of PBSTs after oral administration. The main pharmacokinetic parameters for conventional tablets and multiple oral dose of PBSTs were calculated as the follows: tmax（2.00±0） and （4.00±0） h; Cmax（25.48±0.18） mg/L and （19.24±0.45） mg/L; AUC0-∞（321.42±5.00） mg·h·L-1 and （370.08±12.23） mg·h·L-1. The relative bioavailability of the sustained-release tablets was 119.15％ compared with the conventional tablets. Conclusion PBSTs has the property of sustained-relsease and is bioequivalent to the conventional tablets.

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