Objective To investigate the method of the artificial lipofuscin preparation. Methods Extracting the hepatic mitochondrion of rat, we applied oxidation-reduction method to making and obtaining the artificial lipofuscin.The capability of artificial lipofuscin being digested by peritoneal macrophages of mice was examined. Results The artificial lipofuscin had positive reactions both to specific histochemical stain and FITC fluorescence label.Positive granules or fluorescence were present in cytoplasm of the macrophages and its distribution was similar to that of positive control.Conclusion The artificial lipofuscin had the same histochemical character and biological effect as natural lipofuscin, so it could be used as a substitute for anti-senescence research.

Objective:To investigate the protective effect and potential mechanism of cordycepin on renal proximal tubular cells injury induced by lipopolysaccharide (LPS).Methods:Renal proximal tubular cells NRK-52E were incubated on a cell culture plated at a density of 1×10 5/mL for experiment, then divided into control group (Ctrl group), LPS group (cells were stimulated with 1 mg/L LPS), 10 μmol/L or 20 μmol/L cordycep in intervention groups (LPS+C 10 group and LPS+C 20 group). Cell viability was measured using cell counting kit-8 (CCK-8) reagent. The level of intracellular reactive oxygen species (ROS) was detected by 2',7'-dichlorofluorescin diacetate (DCFH-DA) staining. The protein expressions of inflammatory factors intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), interleukin-1β (IL-1β), and nuclear factor-κB (NF-κB) were detected by Western blotting. Results:Compared with the Ctrl group, LPS significantly inhibited NRK-52E cell viability, increased intracellular ROS, and up-regulated the expressions of ICAM-1, VCAM-1, IL-1β and NF-κB. Compared with LPS group, after treated with 10 μmol/L or 20 μmol/L cordycepin, NRK-52E cell viability was significantly increased (Ctrl group as 1: 0.717±0.017, 0.916±0.036 vs. 0.554±0.046) and intracellular ROS level was significantly decreased (Ctrl group as 1: 1.527±0.165, 1.098±0.168 vs. 2.543±0.127), meanwhile the expressions of ICAM-1, VCAM-1, IL-1β and NF-κB were significantly down-regulated [Ctrl group as 1, ICAM-1/GAPDH: 2.364±0.097, 1.561±0.074 vs. 3.101±0.121; VCAM-1/GAPDH: 2.866±0.135, 1.920±0.098 vs. 4.170±0.119; IL-1β/GAPDH: 2.358±0.107, 1.563±0.179 vs. 3.301±0.210; phosphorylation NF-κB p65 (NF-κB p-p65)/GAPDH: 2.559±0.166, 1.596±0.148 vs. 3.183±0.098], the differences were statistically significant (all P < 0.05). Compared with the LPS+C 10 group, the cell activity of LPS+C 20 group was more significant (0.916±0.036 vs. 0.717±0.017, P < 0.01), and the expressions of ICAM-1, VCAM-1, IL-1β, NF-κB were down-regulated more significantly (ICAM-1/GAPDH: 1.561±0.074 vs. 2.364±0.097, VCAM-1/GAPDH: 1.920±0.098 vs. 2.866±0.135, IL-1β/GAPDH: 1.563±0.179 vs. 2.358±0.107, NF-κB p-p65/GAPDH: 1.596±0.148 vs. 2.559±0.166, all P < 0.05).Conclusion:Cordycepin could significantly increase the survival rate of NRK-52E cells, reduce intracellular ROS level, and inhibit inflammation, and the anti-inflammation effect can be related with NF-κB pathway.

[Objective]To explore the roles of TIDC(tumor infiltrating dendritic cell)and TIL-T(tumor infiltrating T lymphocyte)in tumor immunity of EBV-associated gastric carcinoma(EBVaGC)and their clinical significance.[Methods]TIDC and TIL-T in EBVaGC(n=49)and EBVnGC(EBV-negative gastric carcinoma)(n=20)were detected by immunohistochemistry method and then their subtypes and their relationships with clinical and pathological factors were analyzed.[Results]Mild infiltration was detected in 29 of 49(59.2%)EBVaGCs and 18 of 20(90%)EBVnGCs,while marked infiltration of TIDC was detected in 20 of 49(40.8%)EBVaGCs and 2 of 20(10%)EBVnGCs.And the difference between the two groups was significant (P=0.013).The number of TIL-T in EBVaGC was hisher than that in EBVnGC(P=0.017),and most of them were CTLs.The number of TIL-T was positively correlated with that of TIDC(r=0.386,n=49,P=0.006).The degree of TIL-T infiltration in EBVaGC had negative relationship with the lymph node metastasis(P＜0.001),while TIDC had no relationship with the clinical features.[Conclusion]TIDC and TIL-T may play important roles in the tumor immunity of EBVaGC;TIDC may contribute to the TIL-T recruitment in EBVaGC.

This paper was aimed to study the genetic diversity and genetic relationship of germplasm resource of Lonicerae Japonicae Flos in order to provide references for its breeding. A total of 36 samples from 18 farm varieties and wild species, as well as related species of Lonicera japonica Thunb. from the main production areas were studied by ISSR-PCR markers. The Jaccard coefficient was worked out by NTSYS-pc software. And a cluster dendrogram of different samples was established based on unweighted pair-group method with arithmetic mean (UPGMA). The re-sults showed that 12 ISSR primers generated 129 loci of which 114 loci were polymorphic. The average percentage of polymorphic bands (PPB) is 88.37%. In the cluster dendrogram, different samples of Lonicera japonica are in the same group, which showed that it is a natural species; the wild sample is separated from the cultivated samples; the traditional type-Maohua is relative stable, but the type of Jizhuahua includes complicated varieties, and it has obvi-ous genetic variation; the new variety Jiufeng 1 has already distinct into one stable type. It was concluded that the ISSR method was suitable for germplasm identification, genetic diversity analysis of Lonicerae Japonicae Flos, thus providing a theoretical foundation for its cultivation and breeding.

AIM:To investigate the effect of cigarette smoking condensate on histone deacetylase 2 (HDAC2) and inflammatory mediators in mouse myoblast C 2C12 cells.METHODS:C2C12 cells were treated with cigarette smoke extract (CSE).HDAC2 siRNA was transfected into the cells using Lipofectamine TM 2000.The levels of interleukin-8 (IL-8) and tumor necrosis factor-α( TNF-α) in the culture supernatants were measured by ELISA , and the expression of HDAC2 at mRNA and protein levels was determined by real-time PCR and Western blotting .RESULTS:The expression of HDAC2 at mRNA and protein levels in CSE group was lower than that in control group (P<0.05).The supernatant levels of IL-8 and TNF-αin CSE group were significantly higher than those in control group ( P<0.05 ) .When the cells were transfected with HDAC2 siRNA followed by CSE stimulation , the expression of HDAC2 at mRNA and protein levels was de-creased , and the supernatant levels of IL-8 and TNF-αwere significantly increased as compared with CSE group and control group (P<0.05).CONCLUSION: Under the oxidative stress condition , C2C12 cells generate high levels of IL-8 and TNF-αby down-regulating the expression of HDAC2.

Objective To study on whitening and anti -aging effect of ginseng saponin and its safety.Methods Whitening effect:using vitamin C as the control drug, the inhibition rate of tyrosinase was determined.Anti senescence effect: the aging model of D-was established, and the DPPH was applied to the skin of the rat's neck, and the drug was prepared by 2 times a day.Skin safety evaluation: the skin changes of the skin of the ginseng saponins were observed after the skin was given ginseng saponin and control drugs.Results When the concentration of ginsenoside was 7 mg/mL, the inhibition rate order was, the water solution of vitamin C >ginsenoside nano >ginsenoside aqueous solution ; In the aging model, the surface of the elastic fibers and the skin surface was parallel to the skin, but the elastic fibers were arranged in a more tortuous and non parallel to the skin;DPPH free radical scavenging effect order was, Vitamin C aqueous solution>ginsenoside nano>ginsenoside aqueous solution, and with the dosage of ginsenoside increasing, the efficiency of ginsenoside DPPH scavenging free radical increased significantly;Skin safety evaluation results showed that ginseng saponin nanometer milk smeared skin, no redness, irritation and other phenomena occured.Conclusion Ginseng saponin nanometer milk has obvious whitening effect, and can not cause damage to the skin, is safe and reliable.

Objective:To Investigate a simple, economical and efficient method of primary osteoblast culture and compare their characteristics. Methods:Primary osteoblasts of 1 st -4 th series explant culture from calvarial bones of neonatal Sprague Dawley rats,were collected and osteoblasts shape, mitosis, proliferation, ALP activity and immunohistological expression of osteocalcin and BMP 2 observed. Results:No difference was found in cell shape, spread, conflunce, ALP activity as well as immunohistological stain of osteocalcin and BMP 2 of the osteoblasts, which had been harvested from the 1 st -4 th series explant cultures of Sprague Dawley rats. Conclusion:Series explant culture can harvest the same kind of osteoblasts as the explant culture, and more osteoblasts can be obtained at a single time. It saves money and time and is easy to manipulate.

ObjectiveTo evaluate the application of multi-slice CT (MSCT) perfusion scan technique in predicting renal function recovery after unilateral hydronephrosis treatment.MethodsThirtyeight patients with unilateral obstructive hydronephrosis not shown on intravenous urography (IVU) and a normal contralateral kidney were recruited for this study.Patients were divided into detected (D) and undetected (UD) groups depending on whether the IVU detected urinary tract obstruction.All patients underwent plain abdominal X-ray,gray-scale ultrasonography,excretory urography and MSCT perfusion scan before and after the treatment.Patients were followed-up at six months or more after the treatment for a mean duration of 12.5 months (range from 6 to 22 ).ResultsOf the 38 cases,22 cases were in group D,16 cases were in group UD.On MSCT,renal cortex blood flow (BF) and blood volume ( BV ) value after treatment in group D were 561.1 ± 165.4 ml/( 100 g · min) and 35.9 ± 11.3 ml/100 g compared with before treatment rates of 361.6 ±109.7 ml/(100g· min) and24.1 ±10.2 ml/100g,t=-3.38,-2.34,P＜0.01,0.05.In the UD group,the differences of these parameters were after treatment 38.7 ± 15.4 ml/(100 g · min),10.306 ± 4.925 ml/100 g and before treatment 39.1 ± 22.5 ml/( 100 g · min) and 8.7 ± 4.4 ml/100 g,P ＞ 0.05.In the aspects of BF and BV,there were statistically significant differences between group D and group U D both before and after the treatment,t=9.09,4.15,P ＜ 0.01.ConclusionsM SCT perfusion can provide a valuable prediction technique of the renal function recovery in patients with unilateral obstructive hydronephrosis.Improvement of renal function can be expected after relief of obstructive hydronephrosis if the patients have a BF 361.6 ml/( 100 g · min) and BV 24.1 ml/100 g or greater measured by MSCT perfusion.

OBJECTIVE: To explore the effects of specific immunotherapy by the nasal spray and/or sublingual contain on the regulation the balance of serum Th1/Th2 cell ratio, and the expression of total IgE (tIgE) level in perennial allergic rhinitis patients. METHOD: Thirty-six cases of allergic rhinitis of perennial (PAR) patients with nasal specific immunotherapy (nasal spray group) were chosen as the object of study, 36 PAR patients with sublingual specific immunotherapy (sublingual group) were chosen for the efficacy comparison group, and 32 cases of healthy adults as control (normal control group). The levels of serum IL-2, IL-4, IL-8 and tIgE in PAR nasal spray group and sublingual group were examined by IRMA and double-antibody sandwich assay before and after the treatment of specific immunotherapy (SIT); The content of IFN-gamma was determined by enzyme-linked immunosorbent assay; The infiltration and changes of eosinophilic granulocyte were observed by the smear of nasal secretions. RESULT: In nasal spray group and sublingual group, the contents of serum Th1 cytokines IL-2 and IFN-gamma were reduced levels significantly, while the contents of Th2 cytokines IL-4, IL-8 and tIgE were significantly increased before the treatment of SIT. After the SIT to the maintenance dose, the levels of IL-2 and IFN-gamma in serum were significantly higher than that before treatment, while IL-4, IL-8 and tIgE content were significantly lower in AR patients (P < 0.01 or P < 0.05). The clinical efficacy of nasal spray group and sublingual group were 97.22% and 94.44%, while the two groups was no statistical difference in efficacy (P > 0.05). CONCLUSION With the presence of Th1/Th2 cell ratio and cytokine imbalance, the AR patients manifested as Th2 cell function in accentuation. Nasal mucosa and/or sublingual in the local SIT can have changed no-balance Th1/Th2 cell of by regulating the balance of serum Th1/Th2 cell ratio, the expression level of cytokine expression and the level of tIgE.
Adolescent ; Adult ; Cytokines ; blood ; Female ; Humans ; Immunoglobulin E ; blood ; Immunotherapy ; Lymphocyte Count ; Male ; Middle Aged ; Rhinitis, Allergic, Perennial ; blood ; immunology ; therapy ; Th1 Cells ; cytology ; Th2 Cells ; cytology ; Young Adult

Objective To evaluate the fatty infiltration of lower limbs (pelvis, thighs and calf) by using iterative decomposition of water and fat with asymmetry and least squares estimation‐quantitative fat imaging (IDEAL‐IQ) in Duchenne muscular dystrophy (DMD) patients, and correlate muscle fat fraction(FF) with clinical assessments. Methods The prospective study method was applied, twenty‐one patients with DMD were diagnosed by genetic analysis and/or muscle biopsy and twenty‐one age‐matched healthy control subjects, were evaluated using IDEAL‐IQ, calculating fat fraction (FF) of muscles in the lower limbs. The clinical data of patients, such as age, height, weight, course of disease, time to walk 10 m, Medical Research Council Scale score, were collected. The age and height between DMD group and control group were compared by independent sample t test. The weight, BMI and FF values between two groups were compared by independent samples nonparametric tests (Mann‐Whitney U tests). K related samples nonparametric test (Friedman test) was used for comparison of FF values between pelvic muscles, thighs muscles and calf muscles. The relationships between FF values and clinical assessments were investigated using Spearman correlation test. Results The mean FF of pelvic muscles [13.1(6.9, 33.8)] was significantly higher than that of the thighs muscles [8.3 (23.4, 4.5)] and calf muscles [8.3 (23.4, 4.5)] (all P<0.05) in DMD group. The FF of all the pelvic muscles in DMD were significantly higher than that of the control group muscles (all P<0.05). The gluteus maximus had the greatest mean FF values [34.7 (26.4, 63.7)] in pelvic muscles. The FF of all the thigh muscles in DMD were significantly higher than that of the control group muscles, except for the sartorius, adductor longus and gracilis (Z=0.69, 1.67 and 1.22, respectively, P=0.489, 0.094 and 0.222, respectively). The adductor magnus had the greatest mean FF values [27.2 (12.4, 58.5)] in thigh muscles. The FF of all the calf muscles in DMD were significantly higher than that of the control group muscles, except for the tibialis anterior and tibialis posterior (Z value was1.77 and 1.85, respectively, P value was 0.076 and 0.064, respectively). The gastrocnemius lateral had the greatest mean FF values [7.4 (4.6, 20.9)] in calf muscles. The mean FF value of pelvic muscles (r=0.679, 0.704,?0.761, 0.755, respectively, all P<0.001), thighs muscles (r=0.801, 0.813,?0.738, 0.706, respectively, all P<0.001) and calf muscles (r=0.502, 0.682,?0.637, 0.664, respectively, all P<0.05) were significantly correlate with age, course of disease, %MRC and time to walk 10 m. Conclusions The fat infiltration of lower limbs in DMD patients showes a distinct involvement pattern. IDEAL‐IQ technique is able to quantitatively and objectively evaluate the degree of fat infiltration of lower limbs in DMD, and it has great value in clinical application.