This study was aimed to establish the TLC identification method of Radix Mirab ilis himalaic a. The β-sitosterol and daucosterol were used as the reference substances. The single-factor test was used. A variety of factors which affected TLC were systematically investigated to filter out the best TLC conditions for identification of different batches of medicines. The results showed that the best TLC conditions were as follows: silica gel G plates, extraction solvent (methanol), reagent (5% sulfuric acid in ethanol), extraction method (ultrasonic extraction with methanol), ex-tracted time (30 min), the agent (petroleum ether-ethyl acetate-acetone (5:2:1)) and sample volume (6 μL). It was concluded that the method, which had high separation degree, was reproducible and simple. It can be used as the quality control of Radix Mirab ilis himalaic a.

Objective To compare differences and similarities of the content of trigonelline in Himalaica mirabilis of either wild or cultivated materials from different places. Methods An HPLC method was established to determine the content of trigonelline in Himalaica mirabilis. ZORBAX XDB-CN column (4.6 mm×250 mm, 5 μm) was used, with mobile phase of acetonitrile-0.03%acetic acid (85∶15), flow rate of 0.8 mL/min, detection wavelength of 265 nm, determination wavelength of 360 nm, and column temperature of 30 ℃. Results Regression calculation was made on peak area with the reference solution concentration, and then got the regression equation A=23.409C-26.398, r=0.999 8. Trigonelline showed good linear relation with peak area among the range of 2.004-200.400 μg/mL. The average recovery of trigonelline was 99.57%, RSD=1.11%. Conclusion There was no significant difference in the content of trigonelline of either wild or cultivated materials from different places. This study laid the foundation of application of the cultivated Himalaica mirabilis.

Background Mechanism of dry eye disease has not been established clearly.Some increasing evidences showed that inflammation plays an important role in pathogenesis and development of dry eye disease.However,the association of interleukin-6 (IL-6) with dry eye has not been clarified.Objective This study was to assess the correlation of IL-6 levels in tear and serum of patient with dry eye disease and clinical symptom.Methods Twenty patients with Sj（o）gren syndrome (SS) and 20 cases with non-Sj（o）gren syndrome (NSS) were enrolled from Second Affiliated Hospital of Nanjing Medical University,and 20 healthy volunteers were recruited as the normal controls.Symptom questionnaires were self-answered and multiple dry eye-related clinical tests were performed,including tear film breakup time (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescence staining.IL-6 concentrations in serum and tear were detected by enzyme-linked immunosorbent assay (ELASA).The correlation between IL-6 level and tear film and ocular surface parameters was analyzed by Spearman rank correlation.Written informed consent was obtained from each subject initial of this trial.Results The scores of symptom questionnaires were 18.50 (11.75,23.00),23.00 (19.00,32.00) and 2.00 (0.25,4.00) in the SS group,NSS group and the normal control group,and significantly increasing values were seen in the SS group and NSS group compared with the normal control group (P=0.00,0.00).BUT values in the SS group and NSS group were 2.50 (2.00,3.88) seconds and 3.50 (2.13,4.00) seconds and were lower than 15.00 (13.13,17.00) seconds in the normal control group (P=0.00,0.00).S Ⅰ t values in the SS group,NSS group and the normal control group were 2.00 (1.50,2.88),4.00 (3.00,5.75) and 19.00 (18.00,25.00)mm,showing significant differences between the SS group and the normal control group (P =0.00),the SS group and the NSS group (P =0.00) or the NSS group and the normal control group (P=0.00).The marked elevation of keratoepithelioplasty score was seen between the SS group and the normal control group (5.75 (4.00,7.75) vs.0.00 (0.00,0.75) (P =0.00) or the NSS group and the normal control group (4.50(3.63,6.00) vs.0.00(0.00,0.75)) (P=0.00).Concentrations of IL-6 in serum and tear in the SS group,NSS group were (131.47±21.04) μg/L and (77.81 ± 15.68) μg/L and were significantly higher than (31.62±8.57) μg/L of the normal control group,and those in the tear were (33.44±8.01) μg/L,(18.78 ±5.73) μg/L and (8.77 ± 3.43)μg/L,respectively in the three groups,with a significant reduce in the normal control group (all P=0.00).tL-6 levels in serum and tear were positively correlated with the score of symptom questionnaires or keratoepithelioplasty and negatively correlated with BUT and S Ⅰ t (all P =0.00).Conclusions IL-6 levels in the tear and serum of the dry eye patient are associated with the severity of the disease.IL-6 levels in serum and tear may be objective,sensitive and reliable indicators of dry eye.

Cytoplasmic transduction peptide (CTP) is a newly designed transduction peptide, by which special molecules can be carried out and localized into cytoplasmic compartment. Superoxide dismutase (SOD) is a protein that is difficult to go into cytoplasm. In this study, CTP-SOD fusion gene was amplified from human cDNA by PCR, and the active recombinant protein was successfully expressed in Pichia pastoris. HeLa cells pretreated with CTP-SOD showed a significantly improved survival against the pyrogallol-induced oxidative stress, suggesting CTP-SOD could cross the cell membrane more efficiently and protect cells from oxidative stress.
Antioxidants ; pharmacology ; Cytoplasm ; drug effects ; metabolism ; Genetic Vectors ; genetics ; HeLa Cells ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Oxidative Stress ; drug effects ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Signal Transduction ; drug effects ; physiology ; Superoxide Dismutase ; biosynthesis ; genetics ; metabolism

A 57-year-old man was admitted to the hospital for a 7-year progressively spreading plaques involving the entire body surface, and multiple irregularly sized red nodules and infiltrated patches on the face, trunk and limbs. Histopathological examination showed pleomorphic tumor cells diffusely dis-persed throughout the dennis, giving an appearance of low proliferation. Some cells with cytoplasmic pro-cesses appeared multiangular in shape, lmmunohistochemically, tumor cells were negative for CDla or S-100, but positive for CD45, FXIIIa, CDl4, MHC- Ⅱ, CD68 and lysozyme with extracellular interstitial expression. Ultrastructurally, the cells exhibited cytoplasmic processes and irregularly sized nuclei; no Birbeck granules were observed. Vesicules of low electron-density were seen diffusely in cytoplasm and extracellular matrix. The case is herein diagnosed as cutaneous non-Langerhans cell histiocytosis, which presents with a chronically invasive clinical course. These cells may develop from immature dermal dendritic cells.

Objective To study the enhancement patterns of hilar bile duct wall of ischemic-type biliary lesion (ITBL) on contrast-enhanced ultrasound (CEUS).Methods Eighteen healthy subjects,18 orthotropic liver transplantation (OLT) recipients without complications,and 36 patients,which were subdivided into 2 groups according to the final diagnosis:patients with (n =24) and without (n =12)ITBL,were enrolled in this study.The patients without ITBL had anastomotic biliary stricture (n =3),cholangitis (n =4),biliary sludge (n =1),and acute rejection (n =4),respectively.The images of baseline sonography and CEUS were retrospectively analyzed in consensus by 2 readers.The enhancement time and level of hilar bile duct wall,hepatic artery and liver parenchyma were recorded.Results Hilar bile duct wall became enhancing earlier than liver parenchyma in all of 4 groups.During arterial phase,hyper-or isoenhancing bile duct walls were present in most cases in the groups of healthy subjects,OLT recipients without complications and patients without ITBL.However,non-or hypo-enhancement of hilar bile duct wall were present in 16 (66.7％) ITBL patients,which is different from the other groups (P ＜0.05).Conclusions The main features of ITBL differing from the other groups were non-or hypo-enhancement of hilar bile duct wall in arterial phase.It may be a diagnostic index to apply in detecting ITBL with CEUS.

Objective To evaluate the role of emergency and staged hepatectomy in peritoneal metastasis associated with ruptured hemorrhage of resectable hepatocellular carcinoma (HCC) patients,and investigate the impact of surgery timing-selecting on peritoneal metastasis of postoperative HCC patients.Methods A retrospective analysis was conduct on the pooled data from 38 HCC patients with spontaneously ruptured hemorrhage treated in our hospital from August 2011 to January 2016.These patients were divided into emergency group who underwent hepatectomy within 24 hours at admission,and staged group who underwent the procedure one week after admission.Perioperative events,overall survival (OS) and disease-freesurvival (DFS) rates,incidence of recurrent and metastatic disease were compared between the two groups.Results The perioperative blood loss and transfusion were much more in emergency group than staged group (both P ＜ 0.05).Nevertheless,the incidence of postoperative mortality was not significantly different (6.0％ vs 0％,P ＞ 0.05).The median survival was 22.5 months in emergency group versus 14.2 months in staged group.The 6-month,1-year,3-year OS rates in emergency group were 88.2％,82.4％ and 30.3％ respectively,and 6-month,1-year,3-year DFS rates were 81.3％,54.7％ and 27.3％.The 1-year OS and 6-month DFS rates were higher than those of staged group (both P ＜ 0.05).The incidence of peritoneal metastasis in staged group was higher than that in emergency group,but it was not significantly different (38.1％ vs 29.4％,P ＞ 0.05).Univariate and multivariate analysis indicated that tumor diameter ≥ 10 cm and AFP ＞ 10 000 μg/L were the risk factors for peritoneal metastasis after hepatectomy for HCC patients with spontaneously ruptured hemorrhage.Conclusions Emergency hepatectomy would warrant a better short-term prognosis compared with staged hepatectomy for the HCC patients with spontaneously ruptured hemorrhage.Staged hepatectomy would not raise the possibility of postoperative peritoneal metastasis.The predictors of tumor diameter ≥ 10 cm and AFP ＞ 10 000 μg/L were risk factors for peritoneal metastasis after hepatectomy for spontaneously ruptured HCC patients.

Objective:To investigate the association between polymorphisms of FGFR2 and the susceptibility of breast cancer in Han population in Guizhou province.Methods:Genotyping was performed using PCR-sequence-specific primers(PCR-SSP)in 106 histologically confirmed breast cancer cases and 116 cancer-free controls.Results:The genotype frequencies of rs1219648 TT,TC,and CC were 50%,25.47%.and 24.53% in breast cancer cases and 29.31%,48.28%,and 22.41% in the controls.The gene frequencies of T in breast cancer cases and the controls were 62.74% and 53.45%.respectively.The gene frequencies of C were 37.26% and 46.55%.respectively.The distribution of allele and genotype frequencies of FGFR2 rs1219648 was statistically different between breast cancer cases and the controls(P<0.05).Conclusion:FGFR2 rs1219648 polymorphism influences the susceptibility of breast cancer.TT genotype might serve as a risk factor for breast cancer.

Objective To investigate the ostengenie potential of adipose-derived stem cells(AD- SCs)when exposed to adenovirns containing hBMP-2 cDNA(Adv-hBMP-2)and offer a choice of cell source for gene therapy and tissue engineering.Methods Human adipose tissues were obtained from patients who received orthopaedic surgery or liposuction.ADSCs were obtained by digesting the adipose tissues.Firstly,flowcytometric analysis was performed for the confirmation of mesenchymal stem cell ori- gin and the surface markers including CD34,CD44,CD68,CD71,CD90,and CD105.The ADSCs were transfected by Adv-hBMP-2 and the effects were tested in vitro,lmmunoprecipitation and Western blotting and ELISA were performed for confirming BMP gone transduction and its stable expression.The transform of ADSCs was assessed by extracellular ALP staining,intracellular ALP spectrophotometry,von Kossa staining and RT-PCR.In the in vivo experiment ADSC-Adv-hBMP-2 cells were injected into the hind limb of nude mice and analyzed radiographically and histologically.Results ADSCs were successfully isolated from human adipose tissues.The isolated ADSCs expressed CD44,CD71,CD90 and CD105 and CD34 and CD68 were absent.The result confirmed the mesenchymal stem cell origin of the cells.West- ern blotting and ELISA confirmed successful and persistent hBMP-2 production by ADSC-Adv-hBMP-2 cells.Extracellular ALP staining,intracellular ALP spectrophotometry,yon Kossa staining and RT-PCR revealed that ADSCs treated with Adv-hBMP-2 had a tendency of transfering into osteoblast.X-ray and H&E sections from hind limb of nude mice injected with ADSC-Adv-hBMP-2 cells confirmed bone forma- tion at 2 weeks.Conclusions Liposuction aspirates contain abundant ADSCs that can be transduced with hBMP-2 gene,and the tranduced ADSCs differentiate into the osteoblast.ADSCs may be an ideal source of mesenchyme-lineage stem cells for gone therapy and tissue engineering.

The mechanism of biological actions of quercetin was studied by using metabolomic method and biomolecular network. HPLC-MS was used to analyze the serum metabolome in rats of blank group and quercetin administration group rats, and MS data were processed by MATLAB software. With multivariate statistical analysis of serum metabolite profiles, a clear separation among blank group and quercetin administration group was achieved, potential biomarkers were selected according to the parameters of variable importance in the projection (VIP) and identified according to MS information and database retrieval. Four compounds, related enzymes, action targets and metabolic pathways had been confirmed, namely retinoic acid and RARbeta, arachidonate and COX-2, 3, 5-diodotyrosine and TPO, uridine diphosphate glucose and PDEs. The mechanism of quercetin enhancing ability of retinoic acid on the induction of RARbeta, activating TPO, using as COX-2 and PDEs inhibitor was approved by biomolecular network and related literatures. In this study, a mechanism of multiple biological actions of quercetin was evaluated at the level of the biomolecular network, metabolomics and biomolecular network can be used to investigate the biological effects mechanism of quercetin, which provided a new method to further revealing mechanism of drug action.
Animals ; Antioxidants ; pharmacology ; Biomarkers ; blood ; Chromatography, Liquid ; Mass Spectrometry ; Metabolic Networks and Pathways ; Metabolome ; Metabolomics ; Multivariate Analysis ; Quercetin ; pharmacology ; Rats