The biological effects mediated by vitamin D and vitamin D receptor (VDR) are involved in the regulation of multiple pathophysiologic processes,including calcium phosphorus metabolism,immune regulation,anti-inflammation,anti-infection and cancer prevention,etc.Autophagy is a eukaryotic cell lysosome-mediated catabolic pathway,which is classified into 3 different types:macroautophagy,microautophagy,and chaperone-mediated autophagy.Since autophagy is vital to maintain energy and metabolism in cells,autophagy dysfunction is closely associated with various pathological processes such as inflammation,infection and tumor.Studies have shown that vitamin D/VDR can affect the different stages of autophagy and regulate inflammation in autophagy.Moreover,vitamin D/VDR and autophagy play an important role in autoimmune disease,diabetes,cardiovascular disease,cancer and other inflammatory diseases.

Background Lasein situ keratomileusi(LASIK) imainstream surgery forefractive correction,and femtosecond laseimuch often used to create thin corneal flap.The measuremenof OPTOVUE RTVue-100 OCto flap and stromal bed thicknesseofferuseful basifoLASIK.Ican be used in measuring the thicknesand shape of the corneal flap.Buthe study on the comparison of flap thicknesbetween WavelighFS200 femtosecond laseand MoriM2 microkeratome 90 μm-knife (Mori90 microkeratome) LASIK by OCilack.Objective The aim of thitrial wato compare the featureof corneal flapcreated by the WavelighFS200 femtosecond laseand Mori90 microkeratome.Methodpiloand prospective study wadesigned.Written informed consenwaobtained from each patienprioto LASIK.Sixty righeyeof 60 patientwith myopiomyopiastigmatism were enrolled in thiclinical trial.The patientwere randomized into the FS200 femtosecond lasegroup and Mori90 microkeratome group with matching demography.RTVue OCwaused to measure flap thicknesusing 10 settingon the 60 eye1 month afteoperation.The featureof the LASIK flapwere analyzed based on the measuring outcomes.ResultThe central flap thickneswa(112±3) μm and the mean flap thickneswa(112 ±3) μm in the FS200 femtosecond lasegroup,which wasignificanlowethan the central flap thicknesa(121±7) μm and the mean flap thicknesa(128±11) μm in the Mori90 microkeratome group respectively (P=0.031,0.030).Corneal flapin the FS200 femtosecond lasegroup showed flashape and thain the Mori90 microkeratome group wameniscushape.The central flap thickneswanoevidently differenfrom thaof peripheral thicknesin the FS200 femtosecond lasegroup (P =0.320).However,in the Mori90 microkeratome group,the central flap thickneswaobviously thinnethan thain the peripheral thicknes(P=0.038).The mean deviation between the actual and predicted flap thicknes(110 μm) wa(3±4)μm in the FS200 femtosecond lasegroup and (17±10) μm in the Mori90 microkeratome group,showing significandifference between them (P =0.009).ConclusionRTVue OCdeterminethathe shape of flapcreated by the FS200 femtosecond laseimore uniform and closeto the expected thicknesof 110 μm than the onecreated by the Mori90 microkeratome.OPTOVUE RTVue-100 OCiuseful tool to evaluate the flap shape and thicknesafteLASIK.

Objective To evaluate the relationship between carotid plaque neovascularization and coronary heart disease using contrast-enhanced ultrasound. Methods We studied carotid plaques in 312 patients with coronary artery disease by contrast-enhanced ultrasound [51 patients with acute coronary syndrome (ACS) and 261 patients with stable coronary artery disease (sCAD) ]. We analyzed sonographic features of each plaque, including the enhancement intensity of plaque (A value), the ratio of plaque to carotid artery lumen in enhancement intensity (Ratio), plaque thickness and plaque echo (soft plaque, hard plaque, mixed plaque, calcified plaque). Results The average thickness of plaque in patients with ACS and in patients with sCAD had no significant difference in statistics [(2.6±0.4) mm vs (2.9±0.8) mm, t=-1.903, P=0.058) ]. The group with ACS:soft plaque 43 (84.3%, 43/51), mixed plaque 8 (15.7%,8/51), no hard plaque and calcified plaque. And the group with sCAD:soft plaque 174 (66.7%,174/261), hard plaque 19 (7.3%,19/261), mixed plaques 16 (6.1%,16/261), calcified plaque 52 (19.9%,52/261). The percentage of soft plaque in the acute coronary syndrome group was significantly higher than that in stable coronary artery disease group (χ2=6.274,P=0.012). The A value and Ratio in patients with ACS were prominently larger than those in patients with sCAD [ (11.3±3.2) vs (8.9±3.3) dB, t=7.150,P<0.01;0.6±0.2 vs 0.4±0.2, t=7.419,P<0.01].Conclusion Carotid artery plaque neovascularization density was significantly higher in patients with ACS than that in patients with sCAD by using contrast-enhanced ultrasound, revealing that the neovascularization density is closely related to clinical symptoms of patients with coronary heart disease.

In order to evaluate the characteristic of porous starch (PS) as the solid dispersions carrier of the total Epimedium flavonoids (TEF), the PS was used. The dissolution of icariin was selected as an indicator to analyze the differences of dissolution between TEF and its solid dispersion. TEF was characterized by scanning electron microscopy (SEM), differential scanning calorimetry (DSC) and X-ray powder diffraction (XRD). Solid dispersion was irregular block and no powder characteristics of TEF and PS could be seen in SEM, DSC and XRD analysis suggested that TEF may be present in solid dispersion as amorphous substance. The dissolution rate of icariin has been improved significantly when the proportion of TEF and PS was 1:2. PS as a traditional solid dispersion carrier is worthy of further study.
Calorimetry, Differential Scanning ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; chemistry ; Epimedium ; chemistry ; Flavonoids ; chemistry ; Porosity ; Solubility ; Starch ; chemistry ; X-Ray Diffraction

Objective To construct a eukaryotic expression plasmid vector encoding Cbl-b gene-specific short hairpin RNAs (shRNAs),and to evaluate its interference effect,so as to lay a foundation for further study on the role of Cbl-b in the immunotherapy of malignant melanoma.Methods According to the sequence of Cbl-b cDNA,4 pairs of shRNAs targeting the Cbl-b gene were designed and synthesized,and then inserted into the plasmid PGPU6/GFP/Neo to construct recombinant plasmids.After identification by DNA sequencing,the 4 shRNA expression vectors were cotransfected into 293T cells with the Cbl-b gene eukarytic expresson plasmid,respectively.The knockdown efficiency of these shRNA expression plasmids on Cbl-b expression was evaluated by real-time (RT) fluorescence-based quantitative PCR and Western blot at 48 hours aftert transfection.Results Sequencing analysis revealed that all the 4 pairs of shRNAs were successfully inserted into the eukarytic expression vector PGPU6/GFP/Neo.As RT-PCR and Western blot showed,all the 4 shRNA-expressing vectors could downregulate Cbl-b expession,and the NO.1 shRNA-expressing vector displayed the strongest interference effect(P ＜ 0.05).Conclusions A eukaryotic expression plasmid vector was successfully constructed for Cbl-b gene-specific shRNAs,and the most effective shRNA was selected in this study.

Objective To investigate the role of methylation of genomic DNA and connective tissue growth factor (CTGF) gene promoter in the pathogenesis of diabetic nephropathy (DN). Methods According to the WHO 1999 guideline for diabetes mellitus diagnosis and classification standard,90 patients diagnosed as diabetes mellitus and 30 healthy volunteers were enrolled in this study.All the participants were divided into diabetes mellitus without DN (DM) group (n=48),DN group (n=42) and healthy control group (n=30) accordingly.DNA was extracted from the peripheral blood and high pressure liquid chromatography (HPLC) was used to measure the overall methylation level of genomic DNA.The methylation status of CTGF gene promoter was determined by PCR and sequencing analysis.Serum CTGF protein level was measured by ELISA.Results The overall methylation level of genomic DNA was 5.23％±0.09％ for DN group,4.71％ ±0.03％ for DM group,and 4.37％±0.01％ for healthy control group,with no significant differences among three groups (all P＞0.05).The CTGF promoter methylation level in DN group (22.02％± 12.90％) was significantly decreased,compared to DM group (49.18％±8.01％,P=0.019) or healthy control group (72.18％±19.30％,P=0.000).Moreover,the serum CTGF protein level in DN group [(193.44±11.90) mg/L] was significantly increased,compared to DM group [(127.65±10.30) mg/L,P=0.031] and healthy control group [(95.84±5.10) mg/L,P=0.001]. Conclusion In DN patients,CTGF gene promoter methylation level is significantly decreased,but CTGF protein level is higher as compared to non-DN patients,which indicates that CTGF gene promoter demethylation may be involved in the pathogenesis of diabetic nephropathy.

Objective To investigate the association between Bsml polymorphism in vitamin D receptor (VDR) gene and diabetic nephropathy in Chinese Han population. Methods PCR-restriction fragment length polymorphism (PCR-RFLP) was used to test the genotype and allele frequency of Bsml in 304 patients with type 2 diabetes mellitus (DM group) and 100 healthy individuals (NC group).The DM group was further divided into non-diabetic nephropathy group (DN0 group,122 cases),minimal albuminuria group (DN1 group,87 cases),and mass albuminuria group (DN2 group,95 cases).Eighty-three DM patients without nephropathy for over 5 years were L-NDN subgroup,and 64 DM patients with nephropathy occurring within the first year were EDN subgroup. Results Genotype and allele frequency of BsmI polymorphism were significantly different between DM and NC group (x2=7.088,P=0.008;x2=5.865,P=0.015).BB+Bb genotype and B allele frequency were significantly higher in DN2 group than those in NC group (x2=14.287,P=0.000;x2=12.621,P=0.000) and DN0 group (x2=8.063,P=0.005;x2=8.173,P=0.004).BB+Bb genotype and B allele frequency were significantly higher in EDN group than those in L-NDN group (x2=7.228,P=0.007; x2=5.853,P=0.016).DN patients with allele B (BB and Bb genotypes)presented higher urinary albumin excretion rates compared with patients without allele B (bb genotype,P＜0.01).The genotype of BsmI was correlated with DN,and allele B was risk factor of DN occurrence and early onset (OR=2.004; OR=2.394). Conclusion VDR gene BsmI polymorphism is associated with DN,and the patients carrying allele B are more involved in mass albuminuria and eady onset of nephropathy.

OBJECTIVETo study the proliferation and differentiation of neural stem cells in the subventricular zone (SVZ) in neonatal rats after bilateral common arteries occlusion.

METHODSNinety-six 3-day-old Sparuge-Dawley rats were randomly divided into two groups: ischemia and control. Rats in the ischemia group were subjected to bilateral common arteries occlusion and the rats in the control group were sham-operated. All rats were administrated with 5-bromodeoxyuridine (BrdU) (50 mg/kg) via intraperitoneal injection. Rats were sacrificed and their brains were removed 1, 4, 7, 10, 14 and 35 days after ischemia. Using brain paraffin sections and immunofluorescence assays, the number of newborn cells in the SVZ was counted. Newborn neural stem cells and oligodendrocytes in the SVZ were observed, and then double marked with BrdU and nestin or osmium tetroxide (O4).

RESULTSThe number of BrdU+ cells (neural stem cells) in the SVZ in the ischemia group was greater than in the control group 4, 7, 10 and 14 days after ischemia, and reached a peak at 4 days after ischemia (253.1+/- 49.3 vs 133.5+/- 17.7; P< 0.01). By 35 days after ischemia, the number of BrdU+/O4+ cells (oligodendrocytes) in the corpus callosum (56.0+/- 7.2 vs 17.0+/- 6.4; P< 0.01), the septal nuclei (45.0+/- 11.9 vs 20.5+/- 5.0; P< 0.01), the striatum (34.5+/- 4.2 vs 14.5+/- 5.8; P< 0.01) and the olfactory bulb (46.5+/- 6.6 vs 23.5+/- 8.4; P< 0.01) in the ischemia group increased significantly as compared to the control group (P< 0.01).

CONCLUSIONSBrain ischemia can activate the proliferation of neural stem cells in the SVZ and promote neural stem cells differentiation into oligodendrocytes. The immature brain may have the capacity for self-repair after ischemic brain injury.

Animals ; Animals, Newborn ; Brain Ischemia ; physiopathology ; therapy ; Bromodeoxyuridine ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cerebral Ventricles ; physiopathology ; Female ; Male ; Neurogenesis ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation

Objective To evaluate in vitro effects of specific small interfering RNA (siRNA)-silencing of the casitas B-lineage lymphoma b (Cbl-b)gene on immunocompetence of primary murine lymphocytes. Methods Spleens were resected from C57BL/6 mice, and splenic lymphocytes were sterily isolated and cultured in vitro. These lymphocytes were divided into 3 groups: silence group transfected with a Cbl-b-specific siRNA using the EntransterTM-R 4000 reagent, negative control group transfected with a negative control siRNA using the EntransterTM-R4000 reagent, blank control group receiving no treatment. After additional culture for 72 hours, ELISA was performed to measure levels of interferon γ(IFN-γ)and tumor necrosis factor α (TNF-α)in culture supernatants of lymphocytes. In addition, the Cbl-b gene-silenced lymphocytes were co-cultured with B16F10 melanoma cells to evaluate their immunocytotoxic effects on melanoma cells. Results Splenic lymphocytes were successfully isolated from C57BL/6 mice and cultured in vitro, and the Cbl-b-specific siRNA was also successfully transfected into the primary murine lymphocytes and effectively down-regulated the expression of Cbl-b gene in them. Compared with the negative control group and blank control group, the silence group showed significantly increased supernatant levels of IFN-γ and TNF-α(all P < 0.05). The immunocytotoxic effect of lymphocytes on melanoma cells was significantly stronger in the silence group than in the negative control group. Conclusion Cbl-b gene silencing can promote secretion of IFN-γ and TNF-α by murine lymphocytes, and enhance their immunocytotoxic effects on B16F10 melanoma cells in vitro.

OBJECTIVETo discuss the efficacy of the myogenic elephant skin cream combining with VSD for treating severe bedsores.

METHODSTwenty-nine cases of III and IV degree bedsores were treated from June 2009 to June 2013. Among them, 15 cases were treated by myogenic elephant skin cream combined with VSD (VSD group) including 7 males and 8 females with an average age of (69.0±5.3) years old ranging from 17 to 96 years;other 14 cases were treated by the treatment of conventional dressing change (control group) including 6 males and 8 females with an average age of (71.0±4.2) years old ranging from 40 to 86 years. At 7, 14, 21, 28 d after treatments, specimens of wound two groups were respectively taken to examine immunohistochemical CD34 adopted SABC, the number of wild vascular cross were observed as capillary density value under high magnification microscope. It was used to assess the hyperplasia of granulation tissue of wound. The capillary density value and the visual wound observation were indicators for evaluation of clinical efficacy.

RESULTSSeventeen of 19 cases got complete data of specimens of wound at 7, 14, 21, 28 d,included 9 cases of VSD group,8 of control group. In the microscope view, the capillary density of VSD group was higher than that of control group significantly (P<0.05), it showed the application of VSD technology improved hyperplasia of granulation tissue much faster than conventional dressing change. In VSD group, 13 cases with 15 wounds healed, 2 cases with 3 wounds improved; in control group,3 cases with 3 wounds healed,7 cases with 9 wounds improved, 4 cases with 5 wounds were unhealed.

CONCLUSIONUsing the VSD technology with continuous high pressure suction to clean the drainage of wound and lacuna thoroughly can effectively control infection,promote the growth of granulation tissue, and then applying myogenic elephant skin cream to improve partial blood supply while prompting rapid growth of new granulation tissue and epithelial cell. The risk of this treatment is low,and the course of treatment is short, this provides a safe and effective method for treating bedsores.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Drainage ; methods ; Female ; Humans ; Male ; Middle Aged ; Pressure Ulcer ; therapy ; Skin Cream ; therapeutic use