Objective To compare the transfection efficiency of galactosylated chitosan nanoparticle vehicle with chitosan nanoparticles vehicle,and observe the therapeutic effect of pGL3-hTERTp-TK on HCC cell line HepG2. Methods Preparing the chitosan and galaetosylated chitosan. Constructing the pGL3-hTERTp-TK plasmid and the Ch/DNA and GC/DNA complexes.Transfecting the HepG2 and the normal hepatic cell L-02 with chitosan/DNA and galactosylated chitosan/DNA complexs.Detecting the fluorescence and the expression of luciferase gene using the fluorescent microscope and the scintillation counter.Detecting the cell growth and apoptosis through the Caspase-3 and the flow cytometry. Results Two clear straps appeared in the agarose gel.The locations were 300 bp and 1100 bp.The relative lueiferase activity of pGL3-hTERTp-Luc+mediated by galactosylated chitosan was powerful than which of pGL3-hTERTp-Luc+mediated by chitosan in HepG2 by the scintillation counter.However,the relative luciferase activity was very weak in L-02.The same results were observed by fluorescent microscope.When the concertration of the GCV was 10 μg/ml(t=51.40,P=0.000),the HepG2 cell inhibition which was transfected by GC-pGL3-hTERTp-TK was obviously different from the L-02 cell inhibition which was transfected by GC-pGL3-hTERTp-TK in the statistics.The significant apoptotic rate was 65.28%in HepG2 which was transfected with GC/DNA,whereas it was only 10.80% in L-02. The significant apoptotic rate in HepG2 which was transfected with GC/DNA was very higher than the other groups (LSD, P ＜0.05). The significant apoptotic rate (10.80%) in L-02 which was transfected with GC/DNA was very higher than the group which was Ch-pGL3-control (LSD, P =0.000). The average fluorescence intensity of the HepG2 which is transfected by the Ch-pGL3-hTERTp-TK was 168.02± 3.68. The average fluorescence intensity of the HepG2 which is transfected by the GC-pGL3-hTERTp-TK was 204.45 ±3.45. The two groups had a significant difference in the statistics (t=-12.504, P＜0.05). Conclusion Galactosylated chitosan has higher transfection efficiency than chitosan. GC-pGL3-hTERTp-TK could specially attack HCC cell line and almost has no influence on normal hepatic cells.

Objective:To investigate the association between the C1GALT1 rs1008898,the DEFA rs2738081 polymorphisms and susceptibility to IgA nephropathy in Chinese Han and Hui population in area of Gansu Province and explore molecular markers to predict IgA nephropathy Methods: In this study,there were 146 patients with IgAN and 180 normal controls in Han people and 83 patients with IgAN and 100 normal controls in Hui people.Two SNPs as rs1008898 and rs2738081 were analyzed with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and polymerase chain reaction-gene sequencing technology.The genotype and allele frequency of rs1008898 and rs2738081 were compared between patients with IgAN and normal controls.Results: Rs1008898 GG genotypes and G allele were over represented in IgAN patients compared with controls.Distribution of rs1008898 polymorphism in patients with IgAN and normal controls showed no difference in Hui people.Neither Han nor Hui population,rs2738081 polymorphism had difference between IgAN patients and normal controls.Conclusion: The G allele of rs1008898 probably has correlation with the genetic susceptibility of IgAN in Gansu Han people.

This study is to investigate the protective effect of mangiferin on NF-kappaB (P65) and IkappaBalpha expression in peripheral blood mononuclear cell (PBMC) in rats with cigarette smoke induced chronic bronchitis. The rat model with chronic bronchitis was established by cigarette smoke. Real-time fluorescence RT-PCR was executed for evaluating the NF-kappaB (P65) and IKkappaBalpha gene expression in mononuclear cell, and flow cytometry for their protein expression. The serum hs-CRP (high-sensitivity C-reactive proteins) and TNF-alpha (tumor necrosis factor-alpha) were detected by enzyme-linked immunosorbent assay. The histopathological score was obtained from lung tissue HE staining slides of lung tissue. The results showed that mangiferin could markedly suppress the NF-kappaB (P65) mRNA and protein expression in mononuclear cell, while promote the IkappaBalpha mRNA and protein expression. Furthermore, mangiferin could lower serum hs-CRP and TNF-alpha level, and reduce the chronic inflammatory damage of bronchiole. These results suggested that mangiferin could notably ameliorate chronic bronchiole inflammation induced by cigarette smoke, and this protective effect might be linked to the regulation of NF-kappaB (P65) and IkappaBalpha expression in mononuclear cell.
Animals ; Bronchi ; pathology ; Bronchitis, Chronic ; blood ; etiology ; metabolism ; pathology ; C-Reactive Protein ; metabolism ; I-kappa B Kinase ; genetics ; metabolism ; Leukocytes, Mononuclear ; metabolism ; pathology ; Male ; Mangifera ; chemistry ; Plants, Medicinal ; chemistry ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tobacco Smoke Pollution ; Transcription Factor RelA ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; blood ; Xanthones ; isolation & purification ; pharmacology

AIM: To further elucidate the mechanism underlying the protective effect of Chinese herb Fu Sheng powder on vascular dementia. METHODS: Primary passage of neural cells of new born rats were subjected to the ischemia-reperfusion-like injury of hypoxia plus glucose deprivation followed by reoxygenation plus glucose, and the effect of “Fu-Sheng powder” on neural cells was examined. RESULTS: Both 5 hours of “ischemia” and 5 hours of “ischemia” plus 5 hours of “reperfusion” led to severe injury to neural cells, the formation of MDA and intracellular calcium concentration increased significantly, however, the activities of SOD and fluidity of neurons decreased significantly. It was also observed that Fu-Sheng powder could significantly alleviate this injury. CONCLUSION: Fu-Sheng powder had direct protective effect on neurons subjected to iscehmia-reperfusion-likeinjury, which might be one of the mechanisms underlying its therapeutic effect on vascular dementia. [

Background Tumstatin is the most active endogenous angiogenesis inhibitor,which has a marked inhibitory effect on pathological neovascularization,and Tum5 is an angiogenesis inhibitors fragment of fulllength tumstatin.Objective This study was to investigate the effects of adenovirus-mediated overexpression of recombinant Tum5 gene on the proliferation,migration and tube formation of human umbilical vein endothelial cells (HUVECs) in physiological status.Methods The empty adenoviral vector expressing green fluorescent protein (rAd-GFP) and the viral vector expressing recombinant Tum5 gene were constructed.The HUVECs cultured in RPMI1640 medium were divided into normal control group,empty vector group (rAd-GFP group) and Tum5 gene infection group (rAd-GFP-Tum5 group).The rAd-GFP and rAd-GFP-Tum5 adenoviral particles at the density of 1 × 1010/ml were added into the medium to infect the cells for 48 hours.The proliferation of the cells was assayed at 24,48 and 72 hours by cell counting kit-8 (CCK-8) to evaluate the proliferative rate;the migration number of the cells was detected at 48 hours after infection by Transwell chamber;the tube formation number of the cells were detected by Matrigel method.The concentration of vascular endothelial growth factor (VEGF) in cell supernatants was assayed by ELISA at 24,48,and 72 hours following adenoviral infection.Results The cultured cells showed green fluorescence in the rAd-GFP group and rAd-GFP-Tum5 group under the inverted fluorescence microscope,and the infection efficiency of rAd-GFP and rAd-GFP-Tum5 was 55.13％ and 50.31％,respectively.No significant difference was found in cell proliferative rate among normal control group,rAd-GFP group and rAd-GFP-Tum5 group both at 24 and 48 hours after infection (both at P＞0.05),and the cell proliferative rate was significantly lower in the rAd-GFP-Tum5 group than that in the normal control group and rAd-GFP group at 72 hours after infection (both at P＜0.01).The migration number of the cells at 48 hours after infection was 2 260.25-±930.44,2 370.00±441.06 and 723.75± 363.80 in the normal control group,rAd-GFP group and rAd-GFP-Tum5 group,showing a significant difference among the groups (F =8.524,P =0.008),and the migrated cells were evidently decreased in the rAd-GFP-Tum5 group compared with the rAd-GFP group and the normal control group (both at P＜ 0.01).The tube number at 48 hours after infection was 95.67±5.86,88.00±4.58 and 20.67±3.51 in the normal control group,rAd-GFP group and rAdGFP-Tum5 group,showing a significant difference among the groups (F=226.498,P＜0.01),and the tube number in the rAd-GFP-Tum5 group was significantly reduced in comparison with the normal control group and rAd-GFP group (both at P＜ 0.01).The considerably differences in VEGF concentration in the cell supernatants were found in different groups and various time points (Fgroup =73.260,P＜0.01;Ftime =73.477,P＜0.01),and VEGF concentration in the cell supernatants was significantly decreased in the rAd-GFP-Tum5 group compared with the rAd-GFP group at both 48 hours and 72 hours (both at P＜0.01).Conclusions The overexpression of the recombinant Tum5 can inhibit the proliferation,migration and tube formation of the HUVECs in physiological status,which may be associated with Tum-5-mediated down-regulation of VEGF protein in the cell supernatant.

Objective: To investigate the effect of Xileisan temperature-sensitive gels on endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor A (VEGF-A) and tumor necrosis factor-α (TNF-α) expression in rats with bleeding internal hemorrhoids, so as to provide insights into the illustration of the pathogenesis of internal hemorrhoid hemorrhage. Methods: Thirty six-week-old SPF-graded rats of the SD strain were randomly divided into the normal group, model group and Xileisan temperature-sensitive gel group, of 10 rats in each group (half male and half female). Cotton balls were soaked with 0.16 mL of croton oil mixture and then inserted into the anus of rats in the model group and Xileisan temperature-sensitive gel group for 10 s. After 6 h when the rectal mucosa tissues presented remarkable swelling, the perianal mucosa was rubbed repeatedly with a rough glass rod until the glass rod was bloody. Following successful modeling, rats in the Xileisan temperature-sensitive gel group was given rectal administration of Xileisan temperature-sensitive gel at a dose of 0.5 mL/d, while animals in the normal group and model group were given rectal administration of the blank gel at the same dose. Following administration for 7 successive days, rats were sacrificed, and the hemorrhoids tissues were collected for pathological examinations. The eNOS, VEGF-A and TNF-α expression was determined using immunohistochemistry and compared among groups. Results: Compared with the normal group, the rat hemorrhoids mucosa showed inflammatory changes in the model group, with submucosal congestion and edema, blood vessel congestion and dilation, and visible new blood vessels, and remarkable improvements were seen in the hemorrhoid mucosal inflammation in the Xileisan temperature-sensitive gel group. There were significant differences in the integrated option density (IOD) of eNOS and VEGF-A expression in rat hemorrhoids tissues among the three groups (P<0.05), and no gender-specific differences were seen (P>0.05). The IOD values of eNOS (45.84±13.66) and VEGF-A expression (45.89±9.06) were higher in rat hemorrhoids tissues in the model group than in the normal group (23.11±5.64 and 27.91±11.65) and the Xileisan temperature-sensitive gel group (27.41±8.89 and 33.44±6.20) (P<0.05), while no significant differences were detected in the IOD of TNF-α expression in rat hemorrhoids tissues among the three groups (P>0.05). Conclusion Xileisan temperature-sensitive gel may alleviate inflammation and internal hemorrhoids hemorrhage through inhibiting eNOS and VEGF-A expression in rat hemorrhoids tissues.

Abstract Anorectal malignant melanoma (ARMM) is a subtype of malignant melanoma with low incidence and high invasion, and has a poor prognosis due to its hidden location and rapid progression. In terms of pathogenesis, the molecular landscape and potential carcinogenic driver gene characteristics of ARMM are significantly different from those of cutaneous melanoma, manifested by a lower rate of somatic mutations, no ultraviolet exposure-related mutation characteristics, a high incidence of cell structural variation, and high genomic instability. The tumor-driving genomic mutations are mainly KIT, NRAS, and NF1 mutations, and the incidence of SF3B1 mutations is significantly higher than that in other sites of mucosal melanoma. Surgery is still the main treatment for ARMM, while immunotherapy and targeted therapy need further development. This article reviews the characteristics of carcinogenic driver gene mutations and clinical diagnosis and treatment of ARMM, providing a reference for the prevention and treatment of ARMM.

Objective To explore the clinical and neuroimaging features of a frontotemporal dementia with parkinsonism linked to chromosome 17 ( FTDP-17 ) pedigree caused by mutation of microtubule-associated protein tau ( MAPT) gene.Methods The proband and one patient from a FTDP-17 pedigree were assessed through standardized clinical evaluation , neuropsychology assessment , video-electroencephalogrom ,MRI, genetic sequencing , as well as 18 F fludeoxyglucose ( FDG) SPECT for brain metabolism and 11 C 2β-carbomethoxy-3β-( 4-fluoro ) tropane ( CFT ) PET for dopamine transporter ( DAT ) distribution, respectively.Results A FTDP pedigree with 15 patients (6 still alive) was recruited to this study.The proband and one affected patient were genotyped and confirmed as MAPT c .1788T>G mutation. Parkinsonism was the first symptom for both two patients . Personality, speech changes and dementia accompanied with brain atrophy were developed at the later stage in one patient .The 18 F FDG SPECT studies illustrated asymmetric hypometabolism of the temporal , frontal lobes and basal ganglia in two patients . Regarding to the 11 C CFT PET, one affected patient showed asymmetric decreased uptake of tracer in basal ganglia regions.Conclusions FTDP-17 can display a confusingly broad clinical phenotype , with the parkinsonism as the first symptom . Brain glucose metabolism and DAT distribution could be potential biomarkers in early diagnosis of FTDP-17.

By summarizing the recent literatures on brain mechanisms with acupuncture intervention based on blood oxygenation level-dependent (BOLD)-functional magnetic resonance imaging (fMRI), the BOLD-fMRI examination and analysis methods, the points to be acupunctured, the corresponding meridian activation regions, the specific intensity range, functions and indications of the acupoints, the manifestation of 'bi-directional regulation' characteristics, fMRI performance of chronergy, laterality and needling qi of acupuncture were reviewed to provide the ideas for future research in this area.

Objective To study serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) and its correlation with accumulation degree of 99Tcm-MDP on bone and arthritis and to study the role of sICAM-1 in the diagnosis of patients with early RA. Methods Serum levels of sICAM-1 were measured by immunoenzymetic assay (ELISA). 99mTc-MDP scintigraphy was carried out for the same patients. The significant test between serum levels of sICAM-1 of three groups was performed with analysis of variance; Correlations between serum levels of sICAM-1 and index of semiquatitive 99Tcm-MDP were obtained using sperman's ranks correlation method. Results Compared with control group,serum levels of sICAM-1 were significantly higher in standard group (group A) (P<0.01) and non-standard group (group B) (P<0.05). No significant difference was found between group A and B(P>0.05). Serum levels of sICAM-1 were positively correlated with 99Tcm-MDP scintigraphy. Most patients of non-standard group and standard group showed visible medium-high uptake of 99Tcm-MDP. While control group had not obviously uptake of 99Tcm-MDP. Conclusion Serum levels of sICAM-1, semiquatitive 99Tcm-MDP scintigraphy, accumulation degree of radiopharmuceuutial on bone and arthritis are helpful for diagnosis of early rheumatoid arthritis and judge-ment of the clinical status and improvement of the prognosis.