Objective To study the gene expression of Wnt signal transduction pathway in experimental liver fibrosis and to investigate its role in liver fibrosis. Methods Liver fibrosis model was induced with carbon tetrachloride in 8 SD rats. Another 8 healthy rats were served as control. The gene expression in liver tissues of models and controls were examined using real time PCR array. The differential gene expression was identified as either up- or down-regulated 2-fold. The expressions of smooth muscle actin (SMA), Wnt4, Frizzled2 and β-catenin in the tissues were examined by immunohistochemistry and Western blot. Results The examination confirmed that 36 genes were differentially expressed, including 25 genes up-regulated and 11 genes down-regulated. Compared with the controls, the expressions of Wnt4, Wnt5 a and W nt11 were up-regulated more than 13.9-, 16.5-and 2.17-fold respectively, while the expressions of Wntl and Wnt3 were down-regulated more than 2.32- and 2.15-fold respectively in fibrotic liver. Immunohistochemistry and Western blot showed that the expressions of SMA, Wnt4 and Frizzled2 in fibrotic liver were remarkably higher than those in normal controls. While the level of phosphorylated β-catenin was decreased. Conclusion Both canonical and noncanonical Wnt signal transduction pathway may involve in the mechanism of liver fibrogenesis.

Objective To evaluate the prognostic value of dynamic delta model of end stage liver disease(MELD)in patients with decompensated liver cirrhosis.Methods Ninty-seven patients with decompensated liver cirrhosis were enrolled in the study and followed for 1 year followed up.Child-Turcotte- Pugh(CTP)score and MELD score were calculated twice for each patient on the first day of admission and one month later.The difference between two MELD scores represented the delta MELD.The predictive value related with delta MELD,MELD and CTP scores was determined by the area under receiver operating characteristic(ROC)curve.Results Ten patients died within 3 months,whose delta MELD(3.23?2.77) were higher than those of survivors(0.15?0.39)(P

Objective:To compare the difference between the combined diagnostic effect of plasma Septin9(SEPT9) and polyligand Syndecan-2(SDC2) methylation with four serum tumor markers in the diagnosis of colorectal cancer.Methods:In this study, 128 patients who were treated in the affiliated Hospital of Xuzhou Medical University from March to December in 2019 were selected for a case-control study.All the subjects were examined by gastroenteroscopy.According to the pathological results, they were divided into three groups: colorectal cancer group( n=74) and colorectal adenoma group( n=7). The patients with no abnormal or inflammatory polyps or proliferative polyps examined by gastroenteroscopy were taken as the control group( n=47). The methylation levels of SEPT9 gene and SDC2 gene were detected by Roche Lightcycler 480 II real-time fluorescence quantitative polymerase chain reaction, and the concentrations of alpha-fetoprotein, carcinoembryonic antigen, carbohydrate antigen 125 and carbohydrate antigen 199 were detected by Roche Cobas 8000 electrochemiluminescence instrument.Chi-square test was used to compare the positive rate of each marker in the three groups.Medcalc was used to draw the receiver operating characteristic curve (ROC) curve of the subjects′ working characteristic curve, and the value of each index in the diagnosis of colorectal cancer was analyzed. Results:The positive rates of SEPT9 gene and SDC2 gene methylation were 81.1%(60/74) and 67.6%(50/74) respectively in colorectal cancer group, and increased to 85.1%(63/74) after combined detection.The positive detection rates of alpha-fetoprotein, carcinoembryonic antigen, carbohydrate antigen 125 and carbohydrate antigen 199 in colorectal cancer group were 1.4%(1/74), 33.8%(25/74), 6.8%(5/74) and 13.5%(10/74), respectively.When the four tumor markers were detected together, the positive detection rates were only increased to 43.2%(32/74), except for AFP and carbohydrate antigen 125(χ 2=3.847, 2.430, all P>0.05). The differences were statistically significant (χ 2=48.230, 30.487, 43.285, 3.847, 8.788, 6.988, 8.722, all P<0.05). The area under the curve (AUC) of SEPT9 methylation, SDC2 methylation, alpha fetoprotein, carcinoembryonic antigen, carbohydrate antigen 125 and carbohydrate antigen 199 were 0.854 (0.781, 0.910), 0.795 (0.715, 0.861), 0.575 (0.485, 0.662), 0.685 (0.597, 0.764), 0.603 (0.513, 0.689) and 0.631 (0.541, 0.715), respectively.The AUC of combined detection of two DNA methylation markers was better than that of alpha fetoprotein, carcinoembryonic antigen, carbohydrate antigen 125 and carbohydrate antigen 199, and the differences were statistically significant (alpha fetoprotein: Z=4.990, P<0.001; carcinoembryonic antigen: Z=3.743, P<0.001; carbohydrate antigen 125: Z=4.951, P<0.001; carbohydrate antigen 199: Z=3.983, P<0.001). The combined detection of two kinds of gene methylation was better than the combined detection of four kinds of serum markers in the diagnosis of colorectal cancer, and the difference was statistically significant ( Z=3.334, P<0.001). Conclusion:The combined detection of SEPT9 gene and SDC2 gene methylation in plasma is more suitable for non-invasive diagnosis of colorectal cancer than the combined detection of 4 serum tumor markers.

Objective To evaluate the influence of belly beard device and the distended bladder on the dose distribution of PTV and the dose-volume histograms(DVHs)of organs at risk(OARs)for postoperative radiation tIlerapy of rectal cancer.Methods A total of 23 patients(8 and 15)with distended bladder receiving 3-field postoperative radiation therapy were dealed with or without a special belly beard in the prone position.At the same time,15 cages with belly board were scanned with empty bladder.The volume of irradiated small bowel was calculated for doses between 5-50 Gy at 5 Gy intervals.With prescription dose in plan target volume(PTV)of 50 Gy,we compared the dose distribution,DVH of OARs,conformity index(CIPTV),the volume of irradiated small bowel and the acute toxicity under the condition of thlee different moulds.Results There was no significant difference in PTV's converge,DVHs of femoral head and CI among 3 moulds(P＞0.05).With the belly board,the high-dose volume of irradiated small bowel(V20-V52.5)was significantly decreased(P＜0.05),specially with distended bladder.However,the low dose volume(V5-V15)was slightly increased.The bladder distension significanfly decreases the volumes of the irradiated small howel at dose levels from 15-52.5 Gy(P＜0.05).Furthermore,the mean volume(V5-V30)of irradiated small bowel differed significantly between patients experiencing Grade 0.1 and ≥2 diarrhea(P＜0.05).Conclusions The combination of belly board and distended bladder was more effectively to reduce the irradiated small bowel volume among 3 moulds,so as to minimized acute diarrhea toxicity.

Objective To study the effect of magnetic stimulation on the expression of B cell lymphoma/leukemia gene 2 ( Bcl-2 ) and caspase-3 genes, and the apoptosis of neurons in rats with spinal cord injury (SCI).Methods Sixty rats were randomly divided into a magnetic stimulation group, a model group and a sham-operation group. An SCI model was established in the magnetic stimulation and model groups. The magnetic stimulation was applied at the 6th, 12th, 24th and 72nd hour after the operation to the rats in the magnetic stimulation group, and sham magnetic stimulation was given to the model group and sham-operation group rats at the same time points. Two hours after treatment, 5 rats of each group were sacrificed and their injured spinal cords were sectioned. The gene expressions were detected using immunohistochemical techniques, and apoptosis of neurons was observed by the TUNEL method. Results Few apoptotic cells were found in the sham-operation group, but more were found in the model group. Apoptotic cells in the magnetic stimulation group were significantly fewer than in the model group. The expression of both Bcl-2 and caspase-3 in the magnetic stimulation and model groups was significantly higher than in the sham-operation group at the different time points. Expression of Bcl-2 in the magnetic stimulation group was significantly higher than in the model group, but expression of caspase-3 in the magnetic stimulation group was significantly lower than in the model group. Conclusions Magnetic stimulation up-regulates the expression of Bcl-2 genes and down-regulates the expression of caspase-3 in injured neurons. Magnetic stimulation might have protective and rehabilitative effects after human SCI.

Objective To investigate the effects of lanthanum chloride on proliferation and migration activity of human cervical cancer cells in vitro which may be a new anti-cervical cancer drug and provide experimental data for cervical cancer treatment. Methods HeLa cells cultured in vitro were divided into two groups: experimental group and control group. In experimental group, the cells were respectively treated with lanthanum chloride at different concentrations, 5, 50 and 100 μmol/L, while the cells in the control group were not treated with lanthanum chloride. The cell growth was observed by inverted microscope and the morphology changes of the cells were observed by the laser scanning confocal microscope (LSCM).Proliferation of HeLa cells in the two groups was detected by methyl thiazolyl tetrazolium (MTT) test;apoptosis rate was analyzed by flow cytometry (FCM). Cell migration test was applied to observe the effect of lanthanum chloride on migration. Reverse transcription (RT)-PCR was employed to evaluate the effects of lanthanum chloride on proliferation gene (cyclinD1), anti-apoptosis gene (zinc finger protein A20) and migration-related gene (matrix metalloproteinase 9, MMP-9). Results The status of cell growth was observed under the inverted microscope: with the increased of the lanthanum chloride concentrations, the cell density of reduced, the granule in cytoplasm increased, color intensifying and intercellular space enlarged; some cells became rounding and dead, floating in the culture media; the exfoliated cells increased gradually in the experimental groups. While In the control group, the cells grew adherently, with clear morphology and plump cytoplasm, and adjacent cell grew in lamellar. Observed with LSCM: the nuclear chromatin condensated and marginated with the volume of nuclear decreased in experimental groups. With the increase of the lanthanum chloride concentrations, nuclei in the experimental groups became pyknotic and then underwent karyorrhexis. However, the nuclear of the cells in control group were inact. The growth inhibition rates of lanthanum chloride groups (5, 50, 100 μmol/L) were 24%, 51% and 78%,respectively, in which each was significantly higher than that of the control group (P < 0. 05); the apoptosis rates of lanthanum chloride group were (4. 91 + 0. 39) %, (7. 30 + 0. 71) % and (13.48 + 0. 92) %,respectively, which were all significantly higher than that of the control group [(0. 89 + 0. 11) %, P <0.01]. The migration ability of the cells was also decreased by the treatment of lanthanum chloride, the number of migrated cells in lanthanum chloride groups were 22.2±4. 3, 12. 0±3.2 and 7. 8±2. 6 respectively, which were all significantly lower than that of the control group (41.2±5.4, P < 0. 01). The expression of genes of cyclinD1, A20 and MMP-9, were all decreased by the treatment of lanthanum chloride in a dose-dependent manner. Conclusion Lanthanum chloride can inhibit the proliferation and migration of cervical cancer cells, and induce apoptosis by down-regulating cyclinD1, A20, and MMP-9 expressions in vitro.

OBJECTIVE:To investigate the association of synergistic effects of Wuzhi capsules on tacrolimus with CYP3A5*3 (6986A>G,rs776746) gene polymorphisms. METHODS:One hundred and severty patients underwent renal transplantation receiving tacrolimus maintenance therapy after surgery were selected from our hospital during Jan. 1997-Dec. 2015,and then divided into Wuzhi capsules(+)group(74 cases)and Wuzhi capsules(-)group(96 cases)according to the use of Wuzhi capsules. Both groups received tacrolimus+mycophenolate mofetil+prednisone;Wuzhi capsules (+)group was additionally given Wuzhi capsules,one capsule each time,bid,for more than 12 months. Trough concentration of tacrolimus was detected by CMIA 0,1,3,6,12 months after medica-tion,and the blood concentrations(C0/D)were calculated at different time points after correcting daily dose. CYP3A5*3 gene polymor-phisms was detected by PCR-RFLP. The association of C0/D value with gene polymorphism was investigated by analysis of covariance. RESULTS:Among 170 patients,there were 65 cases of CYP3A5 GG genotype,83 cases of AG genotype and 22 cases of AA geno-type;genotype frequencies were 38.2%,48.8% and 12.9%,which was in line with Hardy-Weinberg balance (P>0.05). There was statistical significance in the distribution frequencies of GG,AG+AA genotype between Wuzhi capsules(+)group and Wuzhi capsules (-)group (P<0.05). After 1 month of medication,C0/D of tacrolimus in GG genotype was significantly higher in Wuzhi capsules (+)group than in Wuzhi capsules(-)group. After 1,3,6,12 months of medication,C0/D of tacrolimus in AG+AA genotype was sig-nificantly higher in Wuzhi capsules(+)group than in Wuzhi capsules(-)group,with statistical significance(P<0.05). There was no statistical significance in C0/D of tacrolimus in GG genotype between 2 groups after 3,6,12 months of treatment(P>0.05). CON-CLUSIONS:Wuzhi capsules can increase C0/D of tacrolimus in CYP3A5*3 AG+AA genotype,but have no significant effect on C0/D of tacrolimus in GG genotype;CYP3A5*3 genotype should be considered when using Wuzhi capsules as synergist of tacrolimus.

BACKGROUND: Bone exposed wounds are frequently required to deal with in orthopaedic surgeries, involving the treatment of open fractures, bone tumors, osteomyelitis, and many other diseases, in which the defect of soft tissue caused by open fractures is the most difficult to deal with. Conventional debridement or negative pressure closed drainage technology is difficult to make bone exposed wounds heal, and the process is extremely cumbersome, during which,patients suffer a lot of pain.OBJECTIVE: To evaluate the advantages and disadvantages of the various types of dressings, and review the application of new hydrogel dressing in bone exposed wounds based on its advantages, such as keeping wound environment moisture, restoring skin physical barrier, contributing to routine dressing change.METHODS: A computer-based online retrieval of PubMed and CNKI databases was performed to search papers published between 2000 and 2016 using the key words hydrogel dressing, bone exposed wound, traditional wound dressing, antibiotic in English and Chinese, respectively. A total of 55 papers suitable for final analysis from the application of traditional and new dressings in bone tissue engineering were reviewed.RESULTS AND CONCLUSION: The treatment of bone exposed wounds involves the treatment of many diseases, such as open fractures, bone tumors, osteomyelitis, which is still an orthopedic problem to solve. The novel hydrogel dressings with unique advantages are able to provide better plans for bone exposed wounds, and the use of these dressings solves the regeneration and repair of exposed bone, and improves the infection of antibiosis. In addition, the gel dressings currently have become a hot spot of research because of the characteristics of sustained-release.

Immuno-fluorescence technique can qualitatively determine certain nuclear translocation, of which NF-κB/ p65 implicates the activation of NF-κB signal pathways. Immuno-fluorescence analysis software with independent property rights is able to quantitatively analyze dynamic location of NF-κB/p65 by computing relative fluorescence units in nuclei and cytoplasm. We verified the quantitative analysis by Western Blot. When we applied the software to analysis of nuclear translocation in lipopolysaccharide (LPS) induced (0. 5 h, 1 h, 2 h, 4 h) primary human umbilical vein endothelial cells (HUVECs) , we found that nuclear translocation peak showed up at 2h as with calculated Western blot verification results, indicating that the inventive immuno-fluorescence analysis software can be applied to the quantitative analysis of immuno-fluorescence.
Active Transport, Cell Nucleus ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Fluorescent Antibody Technique ; Human Umbilical Vein Endothelial Cells ; Humans ; NF-kappa B p50 Subunit ; metabolism ; Software

Genetic mutations are important molecular biomarkers for cancer diagnosis and surveillance.Therefore,the development of methods for mutation detection characterized with straightforward,highly specific and sensitive to low-level mutations within various sequence contexts is extremely needed.Although some of the currently available methods have shown very encouraging results,their discrimination efficiency is still very low.Herein,we demonstrate a fluorescent probe coupled with blocker and property of melting temperature discrimination,which is able to identify the presence of known or unknown single-base variations at abundances down to 0.1％ within 20 min.The discrimination factors between the perfect-match target and single-base mismatched target are determined to be 10.15-38.48.The method is sequence independent,which assures a wide range of application.The new method would be an ideal choice for high-throughput in vitro diagnosis and precise clinical treatment.