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Background: Vitiligo is a depigmentation disorder characterized by losing functional melanocytes, leading to skin and/or hair depigmentation. Kaliziri (Vernonia anthelmintica (L.) Willd) seeds have been traditionally used to treat pigmentation disorders in Central Asia. The extracts of these seeds increased melanin content and tyrosinase activity and upregulated melanogenesis-related proteins in B16 melanoma cells. We isolated the main active compound Kaliziri standard Extracts (encoded as CAM-Y7) from the Kaliziri seeds in a previous study. Our goal was to reveal whether CAM-Y7 promotes melanogenesis in melanocytes and restored pigmentation in animal models of vitiligo. Methods: Herein, we analyzed the melanogenic effects of CAM-Y7 on melanocytes and the skin pigmentation of hydroquinone (H.Q.)-induced mouse model and hydrogen peroxide (H2O2)-induced guinea pig model of vitiligo. Results: CAM-Y7 promotes Melanogenesis up-Regulation MITF-induced Tyrosinase Expression via p38MAPK signal pathway in melanocytes. Oral administration of CAM-Y7 progressively darkened the dorsal skin and hair of the C57BL/6 mice and guinea pigs. Lillie staining and hematoxylin-eosin staining further showed that CAM-Y7 induced melanogenesis in the epidermis and hair follicles of the animals. Conclusion: Taken together, these results suggest that CAM-Y7 regulation of melanogenesis may be mediated by the activation of p38 MAPK and restored pigmentation in an HQ-Induced C57BL/6 mouse and H2O2-induced guinea pig models of vitiligo. CAM-Y7 might be valuable and promising therapeutic as an agent for vitiligo. Subsequent clinical experiments using are needed to verify our results.

The combination of molecular crowding and virtual imprinting was employed to develop a cost-effective method to prepare molecularly imprinted polymers. By using linear polymer polystyrene as a macromolecular crowding agent, an imprinted polymer recognizable to punicalagin had been successfully synthesized with punicalin as the dummy template. The resulting punicalin-imprinted polymer presented a remarkable selectivity to punicalagin with an imprinting factor of 3.17 even at extremely low consumption of the template (template/monomer ratio of 1:782). In contrast, the imprinted polymer synthesized without crowding agent, did not show any imprinting effect at so low template amount. The imprinted polymers made by combination of molecular crowding and virtual imprinting can be utilized for the fast separation of punicalagin from pomegranate husk extract after optimizing the protocol of solid-phase extraction with the recovery of 85.3 ± 1.2%.

Meiguihua oral solution (MOS), a classical Chinese medicinal formula, was approved by the China Food and Drug Administration for production. However, the quality evaluation of MOS has not been reported. In this present study, qualitative and quantitative analysis of MOS were conducted by ultra high performance liquid chromatography coupled to quadrupole-orbitrap high resolution mass spectrometry (UHPLC-Q-orbitrap-HRMS) and high performance liquid chromatography-tri-quadrupole linear ion trap mass spectrometry (HPLC-tri-Q-LIT-MS). Totally 46 phenolic compounds (21 flavonoids and 25 tannins) were identified in the MOS, among them 14 polyphenols were not reported in raw plant materials of MOS. The simultaneous quantification of ten compounds including gallic acid, quercetin-3-O-sophoroside, ellagic acid, sophoraflavonoloside, hyperoside, isoquercitrin, avicularin, astragalin, quercitrin and juglanin, which were completed in 16 min in the negative electrospray ionization (ESI) mode under multiple reaction monitoring (MRM) method. Linearity was reached fine determination coefficient (r2 > 0.9995). Precisions, repeatability, stability (inter-day and intra-day), and recovery were validated and the relative standard deviations (RSD) were less than 2.9%, 4.7%, 3.6% and 1.79%, respectively. This result proved the high sensitivity and efficiency of the method. The quantitative and qualitative analysis of MOS would provide the substantial basis for further quality control and medicinal values.

Objective. To prepare and study the distribution of magnetic nanoliposomes of Sesquiterpene-Rich Fraction from Cichorium glandulosum (SRF-MLN) (under magnetic field) in the main organs of mice. Methods. The SRF-MLN was prepared by ethanol injection-sonication precipitation method. The quality of its pharmaceutical properties was investigated by the active ingredient lactucin. The drug concentration and distribution of lactucin in different tissues and organs including whole blood, liver, heart, spleen, lung, and kidney were evaluated with Sesquiterpene-Rich Fraction from Cichorium glandulosum (SRF-LP) as a control. Results. The prepared SRF-MLN was spherical and monodispersed with an average particle diameter of 65 ± 1 nm, encapsulation efficiency of 91.3% ± 1.5% (RSD% = 1.5), and drug loading rate of 4.7% ± 0.3% (RSD% = 0.7). Dispersion coefficient PDI was 0.157 and Zeta potential was −17.5 mV. The lactucin concentration of SRF-MLN after 10 min of intravenous injection in heart, liver, spleen, lung, and kidney was significantly higher than that of SRF-LP group. The AUC0-12h values of liver tissue were obviously higher than other tissues and MRT was significantly prolonged under the action of the magnetic field (p Conclusion. The prepared SRF-MLN can change the distribution of drugs in different tissues and organs, prolonging the action time of the drugs in the body, and it has certain specificity under the action of applied magnetic field.

The aim of this study was to isolate key active compounds from Toona sinensis leaves (TSL) with antioxidant and anticancer activities. Four compounds, quercetin-3- O - α -L-rhamnopyranoside (QLR), kaempferol-3- O - α -L-rhamnopyranoside (KLR), 1,2,3,4,6-penta- O -galloyl- β -D-glucopyranose (PGG) and ethyl gallate (EG), were purified from the ethyl acetate fraction of TSL by column chromatography. Their structures were identified by mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectrometry analyses. All four compounds exhibited strong antioxidant activities (including ABTS radical scavenging activities and ORAC values). PGG showed the strongest antioxidant activity. KLR showed the strongest anticancer activities against human hepatoma HepG2 and human breast cancer MCF-7 cells. QLR, PGG, and EG showed relatively weak anticancer activities. These results suggest that four compounds are key chemical compositions valuable for preparing functional foods in the food industry.

Epidermolytic palmoplantar keratoderma (EPPK) is the most frequent form of such keratodermas. It is inherited in an autosomal dominant pattern and is clinically characterized by diffuse yellowish thickening of the skin on the palms and soles with erythematous borders during the first weeks or months after birth. EPPK is generally caused by mutations of the KRT9 gene. More than 26 KRT9 gene mutations responsible for EPPK have been described (Human Intermediate Filament Database, www.interfil.org), and many of these variants are located within the highly-conserved coil 1A region of the α-helical rod domain of keratin 9. Unfortunately, there is no satisfactory treatment for EPPK. Thus, prenatal molecular diagnosis or pre-pregnancy diagnosis is crucial and benefits those affected who seek healthy descendants. In the present study, we performed amniotic fluid-DNA-based prenatal testing for three at-risk pregnant EPPK women from three unrelated southern Chinese families who carried the KRT9 missense mutations p.Arg163Trp and p.Arg163Gln, and successfully helped two families to bear normal daughters. We suggest that before the successful application of preimplantation genetic diagnosis (PGD), and noninvasive prenatal diagnosis of EPPK that analyzes fetal cells or cell-free DNA in maternal blood, prenatal genetic diagnosis by amniocentesis or chorionic villus sampling (CVS) offers a quite acceptable option for EPPK couples-at-risk to avoid the birth of affected offspring, especially in low- and middle-income countries.

A new isoflavane glycoside with a novel structure that we called cicerarietinuoside A (1) in addition to the three known compounds β-daucosterol (2), adenosine (3), and ent-kaurane diterpene glycoside (4) were isolated from seeds of chickpea. Their structures were established based on broad spectral analysis. Compounds 2 and 4 were obtained from chickpea seeds for the first time.

In this study, "two dimensional" molecularly imprinted solid-phase extraction (2D-MIP-SPE) of semi-preparative grade was constructed to fast purify ellagitannins in pomegranate husk extract with the help of crystallization and reverse-phase liquid chromatgoraphy (RPLC). Ellagic acid and punicalagin imprinted polymers were synthesized in batch mode and two semi-preparative MIP-SPE columns were individually packed. After investigaing "functional complementation", 2D-MIP-SPE was constructed using ellagic acid MIP and punicalagin MIP-SPE as the first and second dimension, respectively. Then, pomegranate husk extract was fast divided into four fractions individually enriching in ellagic acid, granatin A, punicalagin and ellagic acid glucoside by 2D-MIP-SPE. With the aid of crystallization and RPLC, ellagic acid (13.5mg) and punicalagin (53.4mg) were fast obtained in 30min. Ellagic acid glucoside was purified to the purity near 100% with a recovery of 86.1%. Granatin A (92%) was directly obtained by 2D-MIP-SPE with the recovery of 81.8%. All above indicated that 2D-MIP-SPE was highly efficient in natural product purification. The concept of "functional complementation" was expected to be a useful tool in the construction of 2D-MIP-SPE.

Mechanisms underlying female gonadal dysgenesis remain unclarified and relatively unstudied. Whether X-chromosome inactivation (XCI)-escaping genes and microRNAs (miRNAs) contribute to this condition is currently unknown. We compared 45,X Turner Syndrome women with 46,XX normal women, and investigated differentially expressed miRNAs in Turner Syndrome through plasma miRNA sequencing. We found that miR-320a was consistently upregulated not only in 45,X plasma and peripheral blood mononuclear cells (PBMCs), but also in 45,X fetal gonadal tissues. The levels of miR-320a in PBMCs from 45,X, 46,XX, 46,XY, and 47,XXY human subjects were inversely related to the expression levels of XCI-escaping gene KDM5C in PBMCs. In vitro models indicated that KDM5C suppressed miR-320a transcription by directly binding to the promoter of miR-320a to prevent histone methylation. In addition, we demonstrated that KITLG, an essential gene for ovarian development and primordial germ cell survival, was a direct target of miR-320a and that it was downregulated in 45,X fetal gonadal tissues. In conclusion, we demonstrated that downregulation of miR-320a by the XCI-escaping gene KDM5C contributed to ovarian development by targeting KITLG.

The combination of molecular crowding and virtual imprinting was employed to develop a cost-effective method to prepare molecularly imprinted polymers. By using linear polymer polystyrene as a macromolecular crowding agent, an imprinted polymer recognizable to punicalagin had been successfully synthesized with punicalin as the dummy template. The resulting punicalin-imprinted polymer presented a remarkable selectivity to punicalagin with an imprinting factor of 3.17 even at extremely low consumption of the template (template/monomer ratio of 1:782). In contrast, the imprinted polymer synthesized without crowding agent, did not show any imprinting effect at so low template amount. The imprinted polymers made by combination of molecular crowding and virtual imprinting can be utilized for the fast separation of punicalagin from pomegranate husk extract after optimizing the protocol of solid-phase extraction with the recovery of 85.3 ± 1.2%.

Background Cichorium glandulosum Boiss. et Huet is used for treatment of liver disorders, and its effects are attributed to sesquiterpenes. This study aims to investigate the hepatoprotective effects of a sesquiterpene-rich fraction (SRF) from the aerial part of C. glandulosum on carbon tetrachloride (CCl4)-induced acute hepatotoxicity in mice, and on priming with Bacillus Calmette–Guerin (BCG) followed by lipopolysaccharide (LPS)-induced immunological liver injury in mice. Methods SRF was suspended in water and administered to mice at 0.05, 0.10 and 0.20 g/kg body weight for 7 consecutive days. An active control drug (bifendate pills) was suspended in distilled water and administered to mice at 0.40 g/kg body weight for 7 consecutive days. Hepatotoxicity was induced by intraperitoneal injection of 0.1% CCl4 (0.2 mL/mouse) at 13 h before the last drug administration, or by tail intravenous injection of BCG (0.2 mL/mouse) before the first drug administration and LPS (0.2 mL/mouse; 8 μg) at 15 h before the last drug administration. Blood samples and the livers were collected for evaluation of the biochemical parameters of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and total bilirubin (TBIL). Results SRF significantly reduced the impact of CCl4 toxicity. The highest dose of SRF (0.20 g/kg) was the most effective, reflected by significant reductions in the levels of AST (P = 0.001), ALT (P = 0.000) and TBIL (P = 0.009). The serum enzymatic levels induced by BCG and subsequent LPS injection were significantly and dose-dependently restored by SRF, reflected by significant reductions in the levels of AST (P = 0.003), ALT (P = 0.003) and TBIL (P = 0.007) for the highest dose of SRF (0.20 g/kg). Conclusion SRF is hepatoprotective in animal models of chemical and immunological acute liver injury.

To analyze the aberrant der(X) chromosome using conventional and molecular cytogenetic approaches in a fetus of second trimester and to discuss its clinical effect.Conventional cytogenetic procedures (GTG and CBG banding) were performed on cultured amniotic fluid cells. Three-color fluorescence in situ hybridization (FISH) consisting of X chromosome enumeration probes(CEPX), CEPY and Tel Xp/Yp was further performed to study the aberrant der(X) chromosome.Der(X) was a rare X/Y translocation. The final karyotypes of the fetus was designated as: 46,X,der(X)t(X;Y)(p22.3;q11.2). ish der(X)t(X;Y)(p22.3;q11.2)(X/Ypter-, DXZ1+, DYZ1+)mat.The combination of FISH and conventional cytogenetic techniques is a powerful tool to determine derivative chromosome and to offer an accurate genetic counseling. Identification of Xp; Yq rearrangement can help estimate the risk of fetus abnormalities and give a more precise prognosis.

To analyze the numerical aberration rate of X, Y and chromosome 18 in sperms from an oligozoospermic male with mosaic trisomy 18 and to perform preimplantation genetic diagnosis (PGD) for the couple.G-banding and fluorescence in situ hybridization (FISH) were performed on metaphase chromosome. Sperm was analyzed in three-color FISH with a probe mixture containing CEP18, CEPY and Tel Xq/Yq. A healthy man with normal semen parameters was used as control.Significant difference in the rates of disomy for chromosome 18 (0.63% vs. 0.16%) and the gonosomes (0.945% vs. 0.35%) and diploidy (0.87% vs. 0.31%) was found in the spermatozoa between the patient and the control. After four embryos were biopsied in one PGD cycle, two embryos with XY1818 and XX1818 were selected for implanting and clinical pregnancy was ongoing.Sperm-FISH allows further understanding of aneuploidy rate and accurate genetic counseling. FISHPGD was effective for patient with mosaic trisomy 18.

To analyze the sex chromosome meiotic segregation in inv(Y) patients by fluorescence in situ hybridization (FISH).Conventional cytogenetic procedures (GTG and CBG banding) and FISH were performed on metaphase chromosome. Three-color FISH was performed on sperm samples using a probe mixture containing CEPX, Tel Xp/Yp and Tel Xq/Yq to investigate the sex chromosome segregation of five inv(Y) (p11.1q11.2) carriers. A healthy man with normal semen parameters was used as control.There was no statistical difference in the abnormal sex chromosome number and recombination frequencies in each spermatozoon from the patient in comparison with that in the control.There was no apparent sex chromosome abnormality in the sperm of the inv(Y) (p11.1q11.2) carriers. Sperm-FISH allows further understanding of the sex chromosome segregation pattern and an accurate genetic counseling.

To study the purification technology of Rhizoma et Radix Notopterygii and Radix Angelicae Pubescentis in Fufang Xuelian dropping pills by macroporous resin.Taking osthole, isomperatorin as index ingredients, the type of resin sampling amount and elution solvent were decided, and the influence of sample concentration pH of sample and ratio of diameter to height of column to adsorption were studied.HPD400A was chosen to purify, the suitable sampling ratio of resin volume to raw material was 1:2; pH 3.5 (crude drug) and ratio of diameter to height was 1:7; 95% ethanol of the elution solvent was satisfactory eluant for desorption.HPD400A macroporous resin can be used to purify Rhizoma et Radix Notopterygii and Radix Angelicae Pubescentis.

Colon cancer has a high incidence in the world, especially in Western countries, and the incidence is also increasing in China in recent years. To reduce the incidence, it is essential to identify individuals at high risk and to eradicate risk factors. Although colorectal neoplasia is a multi-stage process, hyperproliferation is the main factor leading to the initiation of carcinogenesis. In animal models of colonic cancer, an increased colonic proliferative rate resulted from inherited differences, pharmacological or surgical intervention, which increase the susceptibility of the colonic mucosa to carcinogens. On the other hand, interventions that decrease colonic proliferation are associated with decreased susceptibility to carcinogens. PCNA (proliferating cell nuclear antigen) is an accessory protein of DNA polymerase and is thought to play an important role in the elongation or replication of the DNA chain. Its accumulation in the nucleus during the G-1 and S stages of the cell cycle has been reported, and the labeling index, which is the percentage of PCNApositive cells, has been reported to be correlated with the proliferative activity and the prognosis of various ma lignant tumors. Thus it is valuable to assess what ro le PCNA plays dur ing the transformation of the colonic polyps to colonic cancer.

The human upper gastrointestinal tract is often infected with Helicobacter pylori (H. pylori). This urea splitting bacterium is now considered to be a causal agent in some diseases, including antral gastritis and frank duodenal ulceration, in addition to an association with gastric carcinoma and mucosa associated lymphoid tissue (MALT) lymphoma[1]. Since the discovery of H. pylori, a number of additional Helicobacter species have been isolated from the stomachs and intestinal tracts of a variety of mammalian species. At least eighteen separate Helicobacter species have been recognized (Table ​(Table11)[2,3]. The discovery of these Helicobacter species, has raised the possibility of a relationship between Helicobacter infection and liver diseases[3]. Table 1 Helicobacter species and their hosts HELICOBACTER PYLORI AND PEPTIC ULCER IN CIRRHOSIS Historically, it is well recognized that duodenal ulcer disease is more common in patients with cirrhosis as compared with non-cirrhotic patients[4]. However, a number of early studies suggested that in cirrhotic patients there was no clear relationship between duodenal ulcers and H. pylori infection, suggesting the possibility of other causes[5]. Other studies suggested that H. pylori infection, as measured by IgG, H. pylori serum antibodies, was more common in cirrhotic patients than in non-cirrhotics[6]. A study showed that cirrhotic patients were more likely to have a positive H. pylori ELISA with a negative histologic examination for H. pylori as compared with noncirrhotic patients[7]. Whether H. pylori is a risk factor for peptic ulcer in cirrhosis remains controversial. In a cross-sectional study by Wang et al[3,8], 49 cirrhotic patients underwent upper gastrointestinal endoscopy and 75 controls (healthy examinees) without liver disease were also examined by endoscopy. Thirty (61%) of the 49 cirrhotic patients had peptic ulcers as compared with 24 (32%) of the 75 controls. The frequency of H. pylori in the antrum in the cirrhotic group was significantly lower than in the control group (39% vs 69%). The presence of H. pylori was more frequent in control patients wit h gastric (75%) and duodenal ulcers (95%) than nonulcerous control patients (59%), the difference between patients with and without peptic ulcer (40% vs 37%) was not significant in cirrhotic patients. H. pylori was identified in 40% of the cirrhotic patients with duodenal ulcers as against 95% of controls with duodenal ulcer (P < 0.05). Nevertheless, this difference was not significant among patients with a gastric ulcer between the two groups (40% vs 75%). There was no significant difference in the frequency of H. pylori infection among nonulcerous patients between the cirrhotic and control groups (37% vs 59%). No evidence was found to substantiate an etiologic role of H. pylori in the development of duodenal ulcer in cirrhotic patients[8]. In 153 consecutive patients with cirrhosis, Siringo et al[9] assessed the prevalence of IgG to Helicobacter pylori and compared it with that in 1010 blood donor-residents in the same area and the relationship of IgG to H. pylori with clinical and endoscopic features and with the risk of peptic ulcer. The prevalence of IgG to H. pylori of cirrhosis was significantly higher than in blood donors (76.5% vs 41.8%; P < 0.0005) and was not associated with sex, cirrhosis etiology, Child class, gammaglobulins and hypertensive gastropathy. In both groups, the prevalence of IgG to H. pylori was significantly higher in subjects aged over 40. Multivariate analysis identified high age and males as risk factors for a positive H. pylori serology and no independent risk factors for peptic ulcer. The high prevalence of H. pylori positive serology found in this series was related to age and sex and might also be explained by previous hospital admissions and/or upper gastrointestinal endoscopy. Their results did not confirm the role of H. pylori as a risk factor for peptic ulcer in patients with liver cirrhosis. H. pylori infection is the major pathogenic factor for peptic ulcer disease. Its epidemiology is not fully known; few data are available in patients with chronic liver disease. To investigate the seroprevalence and factors associated with H. pylori infection, a series of studies or liver cirrhosis patients is necessary. Two hundred and twenty consecutive patients were prospectively included in a study aimed to evaluate the effect of dietary intervention on cirrhosis complications and survival. An epidemiological and clinical questionnaire was completed. Sera were obtained and stored at -70 °C until analyzed. They were tested for H. pylori antibodies using a commercial ELISA kit. Eleven of 220 patients had borderline anti-H. pylori-IgG titers. Of the remaining 209 patients, 105 (50.2%) showed positive titers of H. pylori-IgG. Univariate analysis showed that H. pylori infection was more frequent in older patients, those born outside Catalonia, and in patients with a low educational level. Past ethanol consumption and current smoking were correlated negatively with H. pylori infection. Selected age (OR = 3.1, 95%CI: 1.46-6.45), educational level (OR = 2.2, 95%CI: 1.18-4.2) and alcohol consumption (OR = 0.7, 95%CI: 0.45-0.99) as the variables were independently related to H. pylori infection in multivariate analysis. Their conclusions of H. pylori infection in cirrhosis has the same epidemiological pattern as in the general population. Suggestions that the etiology or the severity of the liver disease could be related to H. pylori infection were not confirmed by their study[10].