Anemia, Refractory ; Humans ; Male ; Middle Aged ; Thrombocytosis

Air Pollutants, Occupational ; analysis ; Solvents ; analysis

Adult ; Amides ; adverse effects ; Benzene ; adverse effects ; Burns, Chemical ; pathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Nitro Compounds ; adverse effects ; Young Adult

OBJECTIVE: To test droplet digital PCR for species identification and absolute quantification of biological sample. METHODS: Specific primers and probes for human mtDNA encoding gene ND4 and 16S rRNA were designed, and the species-specificity was assessed on DNA samples derived from human and common animals. To determine the sensitivity and stability of droplet digital PCR for species identification and absolute quantification, gradient dilution series of recombinant plasmid and 16 human DNA samples were analyzed. RESULTS: Human recombinant plasmid FAM (ND4) could be used in detecting the samples of human. And the results of detecting were consistent with all levels of diluted concentrations. Droplet digital PCR was able to detect low and single copy of target DNA. CONCLUSION Droplet digital PCR, with high sensitivity and specificity, is fully amenable for species identification and absolute quantification of biological samples, also it can be applied on routine forensic examination.
Animals ; DNA ; DNA Primers/genetics* ; DNA, Mitochondrial/genetics* ; Humans ; NADH Dehydrogenase/genetics* ; Polymerase Chain Reaction/methods* ; RNA, Ribosomal, 16S/genetics* ; Sensitivity and Specificity ; Species Specificity

Background Retinal neovascularization disease is a common cause of blinding.Retinal neovascularization is related to enhancing proliferation of vascular endothelial cells.So how to inhibit proliferation of vascular endothelial cells is a hot burning issue.p21 is known to be involved in the regulation of cell cycle and therefore inhibit the cell proliferation.However,the relationship of p21 and the proliferation of vascular endothelial cells in retinal neovascularization disease is for further study.Objective The aim of this experiment was to study the proliferation of rhesus retinal vascular endothelial cells(RF/6A) and expression of p21 in RF/6A cells under the hypoxia condition,and discuss their association.Methods The RF/6A cells were cultured and passaged in vitro,then they were randomly divided into normoxia culture group(5％ CO2 +95％ O2) and hypoxia for 1 hour,3,6,12 hours group(1％ 02+5％ CO2 +94％ N2).Flow cytometer(FCM) was used to check the distribution of RF/6A cell cycle in the normoxia culture group and hypoxia for 1 hour,3,6,12 hours groups.MTT assay was used to detect and compare the cell proliferation(A570)among the various groups.The expression of p21 in the cells was analyzed by Western blot.Results FCM showed that the cells proportion of G0/G1 stage was reduced initially and then increased afterward in hypoxia for 1 hour and 3,6,12 hours groups,showing a significant difference among 5 groups (F =20.083,P =0.000),and the cells proportion of S stage and G2/M stage were increased firstly and then declined in different hypoxia groups with statistical significances (F =7.861,P =0.001 ; F =10.305,P =0.003).Compared with normoxia culture group,cells proportion of G0/G1 stage was declined and that of S stage and G2/M stage were raised after hypoxia culture,showing statistically signifcant differences(P＜0.05).MTT showed that cell multiplication capacity(A570 value)strengthened firstly and then weakened in hypoxia groups with time prolongation,showing a significant difference among all the groups(F=7.768,P=0.001),and A570 value in hypoxia for 3 hours and 6 hours groups (0.315± 0.062,0.365 ± 0.064) was significantly higher than that of the normoxia group (0.205 ± 0.063),respectively(P＜0.05).Western blot showed that the expression of p21 in the cells down-regulated at the beginning and then up-regulated with the increase of hypoxia time,and there was statistical significance (F =16.738,P=0.000).The p21 relative levels in different hypoxia groups were reduced in comparison with the normoxia group,showing statistical signifcances(P＜0.05).Conclusions Short-term hypoxia could reduce the expression of the p21 in RF/6A and induce cell proliferation initially,then p21 increases and cell proliferation is inhibited with the prolongation of hypoxia time.

Objective To study the effect of dialysis adcquacy,microinflammation and residual renal function on nutritional status of hemodialysis patients.Methods One hundred and fourteen patients were enrolled in this study.Kt/V,?_2-MG and serum iPTH were measured as markers of hemodialysis adequacy.Nutritional evaluation included MQSGA,Alb,Hb,TF,IGF-1,IGFBP-3 and anthropometrics such as HGS,BSF,TSF,MAC,MAMC and AMA.Serum IL-6,TNF-?and CRP were detected to assess microinflammation.Urinary volume of 24 hours was measured to investigate the residual renal function (RRF).Results (1)There were different correlations and regressive associations of Kt/V,iPTH and?_2-MG with HGS,MAMC,AMA,Alb,Hb,nPCR,IGF-1 and MQSGA respectively.(2) There were significant correlations and regressive associations of RRF to HGS,TSF,MAMC,Alb,nPCR and IGF-1 within the first year of hemodialysis.(3) There were different correlations and regression relationships of IL-6,TNF-?and CRP with HGS、MAMC、AMA、Alb、TSF、Hb、nPCR、IGF-1 respectively.(4) Multivariate analysis showed that Kt/V,iPTH,IL-6, TNF-?,?_2-MG and RRF were influencing factors,among them,Kt/V,iPTH,IL-6 and TNF-?were independent predictors of nutritional status.Conclusions Hemodialysis adequacy and micruinflammation may impact on nutritional status.Residual renal function may be involved in nutritional status in the first year of hemodialysis.Kt/V,iPTH,IL-6 and TNF-?are independent factors affecting nutritional status.

1,3,5-Trimethylbenzene (1,3,5-TMB) was used as the pore-enlarging modifier to expand the pore size of MCM-41 (mobil company of matter) mesoporous silica nanoparticles. The solvent impregnation method was adopted to assemble non-water-soluble β-carotene into the pore channel of MCM-41. The MCM-41 and drug assemblies were characterized by TEM, FT-IR, elemental analysis and N2 adsorption-desorption. The results showed that MCM-41 has good sphericity and regular pore structure. The research also investigated the optimal loading time, the drug loading and the vitro stability of the β-carotene. As a drug carrier, the modified MCM-41 showing a shorter drug loading time, the drug loading as high as 85.58% and the stability of β-carotene in drug assemblies has improved. The study of this new formulation provides a new way for β-carotene application.
Drug Carriers ; chemistry ; Drug Delivery Systems ; Drug Stability ; Nanoparticles ; chemistry ; Silicon Dioxide ; chemistry ; beta Carotene ; chemistry ; pharmacology

OBJECTIVETo explore therapeutic effect and action mechanism of regulating spleen-stomach needling on diabetic nephropathy (DN).

METHODSUsing multi-centric, randomized, controlled and blind principles, 144 cases of DN were divided into an observation group and a control group according to random digital tab, 72 cases in each one. Based on regular treatment of diabetes, the regulating spleen-stomach needling was applied at Zhongwan (CV 12), Quchi (LI 11), Hegu (LI 4) and Xuehai (SP 10), etc. in the observation group while Shenshu (BL 23), Taixi (KI 3), Sanyinjiao (SP 6), Yanglingquan (GB 34), etc. were selected in the control group by reference of Acupuncture and moxibustion. The treatment was given twice a day, six days as a treatment session with interval of one day between sessions. Totally six weeks were required. Changes of clinical symptoms and signs, fast blood glucose (FBG), urinary albumin excretion rate (UAER), beta2-microglobulin (beta2-MG), monocyte chemotactic protein-1 (MCP-1), lymphocyte membrane cholesterol, propanediol (MDA), PCO, 8-hydroxydeoxy guanosine (8-OHdG), superoxide dismutase (SOD), CD3+, CD4+, CD8+, and CD4+/CD8+ were observed before and after treatment in two groups.

RESULTSAs for improving clinical symptoms and signs, total effective rate was 84.29% (59/70) in the observation group and 55.56% (40/72) in the control group, which had statistical difference between two groups (P<0.01). As for regulating glycometabolism [(6.25 +/- 0.32) mmol/L vs (8.09 +/- 0.63) mmol/L], reducing UAER [(154.43 +/- 55.14) mg/24h vs (268.91 +/- 77.65) mg/24h], restraining over-expression of MCP-1 [(137.59 +/- 36.15) pg/mL vs (166.89 +/- 42.82) pg/mL], regulating level of oxidative stress, prohibiting oxidation of protein and adjusting quantity and activity of T lymphocyte subgroup, the observation group was superior to the control group (P< 0.05, P<0.01).

CONCLUSIONThe regulating spleen-stomach needling is an effective method for treatment of DN, which cold improve glycometabolism disturbance-induced progressive kidney injury, recover glomerular filtration, reduce urinary albumin excretion rate, restrain overexpression of MCP-1, adjust level of oxidative stress, prohibit oxidation of protein, increase protectiveness of membrane, adjust quantity and activity abnormity of T lymphocyte subgroup, leading to repairing lymphocyte damage and improving immune expression to delay kidney damage.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Diabetic Nephropathies ; immunology ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Spleen ; physiopathology ; Stomach ; physiopathology

A novel drug delivery system combining oral fast dissolving film with sodium deoxycholate/phospholipid mixed micelles was prepared to increase the absorption of cucurbitacin B that is a poor aqueous solubility substance. Encapsulation efficiency, particle size, zeta potential, polydispersity coefficient, investigated the morphology, disintegration time of oral fast dissolving film and the pharmacodynamic properties of cucurbitacin B sodium deoxycholate/phospholipid-mixed micelles before and after solidified in mice were evaluated and compared. The oral fast dissolving film prepared in this study showed a homogeneous pale yellow and could completely disintegrated in the 30 s. It could meet the requirements of rapidly disintegrating fully. The encapsulation efficiency, particle size, zeta potential, polydispersity coefficient of cucurbitacin B sodium deoxycholate/phospholipid-mixed micelles loaded in oral fast dissolving film were (43.36 +/- 2.12)%, (108.82 +/- 5.2) nm, (-34.18 +/- 1.07) mV, 0.088 +/- 0.012, respectively. The encapsulation efficiency, particle size, zeta potential, polydispersity coefficient of cucurbitacin B sodium deoxycholate/phospholipid-mixed micelles in solution were (41.26 +/- 2.22)%, (181.82 +/- 4.48) nm, (-30.67 +/- 0.81) mV, 0.092 +/- 0.012, respectively. The difference of pharmacodynamics among film of cucurbitacin B-loaded micelles, cucurbitacin B-loaded micelles and free cucurbitacin B in vivo was compared. Solubility of cucurbitacin B loaded in sodium deoxycholate/phospholipid-mixed micelles has also been greatly improved. The tumor inhibition rate of cucurbitacin B loaded in sodium deoxycholate/phospholipid-mixed micelles was significantly improved and did not change significantly before and after solidified. These showed that the sodium deoxycholate/phospholipid-mixed micelles could enhance the antitumor activities of cucurbitacin B and the stability of cucurbitacin B sodium deoxycholate/phospholipid-mixed micelles was improved significantly after solidified by oral fast dissolving film technology without pharmacodynamic properties changed significantly.
Animals ; Antineoplastic Agents ; administration & dosage ; chemistry ; Cell Line, Tumor ; Deoxycholic Acid ; chemistry ; Drug Carriers ; chemistry ; Humans ; Male ; Mice ; Neoplasms ; drug therapy ; Phospholipids ; chemistry ; Solubility ; Triterpenes ; administration & dosage ; chemistry

Objective To explore the roles of clara cell secretory protein(CCSP)and eosinophil cationic protein(ECP)in the pathoge-nesis of bronchial asthma and to evaluate their diagnostic value in asthmatic children.Methods Induced sputum samples were obtained from 31 asthmatic children during chronic persistent period and clinical remission period.According to global initiative for asthma(GINA),the total of 31 cases accepted systemic treatment by inhaling glucocorticoid.The patients included 18 boys and 13 girls aged from 3.7 to 12.0 years,and their average age was 7.6 years.Sputum CCSP concentrations were measured by enzyme-linked immunosorbent assay(ELISA).And the concentrations of sputum ECP were determined with Pharmacia UniCAP system.Results Asthmatic children had significantly lower CCSP levels in sputum during chronic persistent period compared with clinical remission period(P