Mechanisms of Ganweikang Tablets against Chronic Hepatitis B: A Comprehensive Study of Network Analysis, Molecular Docking, and Chemical Profiling.

The hepatitis B virus (HBV) is one of the major viral infection problems worldwide in public health. The exclusive proprietary Chinese medicine Ganweikang (GWK) tablet has been marketed for years in the treatment of chronic hepatitis B (CHB). However, the pharmacodynamic material basis and underlying mechanism of GWK are not completely clear. This study is aimed at investigating the pharmacological mechanism of the GWK tablet in the treatment of CHB. The chemical ingredient information was obtained from the Traditional Chinese Medicine Database and Analysis Platform (TCMSP), Traditional Chinese Medicines Integrated Database (TCMID), and Shanghai Institute of Organic Chemistry of CAS. Ingredients and disease-related targets were defined by a combination of differentially expressed genes from CHB transcriptome data and open-source databases. Target-pathway-target (TPT) network analysis, molecular docking, and chemical composition analysis were adopted to further verify the key targets and corresponding active ingredients of GWK. Eight herbs of GWK were correlated to 330 compounds with positive oral bioavailability, and 199 correlated targets were identified. The TPT network was constructed based on the 146 enriched targets by KEGG pathway analysis, significantly associated with 95 pathways. Twenty-five nonvolatile components and 25 volatile components in GWK were identified in UPLC-QTOF/MS and GC-MS chromatograms. The key active ingredients of GWK include ferulic acid, oleanolic acid, ursolic acid, tormentic acid, 11-deoxyglycyrrhetic acid, dibenzoyl methane, anisaldehyde, wogonin, protocatechuic acid, psoralen, caffeate, dimethylcaffeic acid, vanillin, ß-amyrenyl acetate, formonentin, aristololactam IIIa, and 7-methoxy-2-methyl isoflavone, associated with targets CA2, NFKB1, RELA, AKT1, JUN, CA1, CA6, IKBKG, FOS, EP300, CREB1, STAT1, MMP9, CDK2, ABCB1, and ABCG2.

Cloning and characterization of defense-related genes from Populus szechuanica infected with rust fungus Melampsora larici-populina.

Characterization of defense-related genes is critical for breeding disease-resistant poplar varieties and for better management and control of leaf rust disease. In the present study, full-length cDNAs of five Populus szechuanica defense-related (PsDR) genes, pathogen-related protein 1 (PsPR1), ß-1,3-glucanase (PsGns), thaumatin-like protein 1 (PsTLP1), thaumatin-like protein 2 (PsTLP2), and phenylalanine ammonia-lyase (PsPAL), were cloned from the leaves of P. szechuanica infected with Melampsora larici-populina (MLP). PsPR1 (728 bp), PsGns (1189 bp), PsTLP1 (929 bp), PsTLP2 (885 bp), and PsPAL (2586 bp) were predicted to encode 161, 347, 245, 225, and 711 amino acid residue-containing proteins with isoelectric points of 8.53, 4.96, 4.51, 7.32, and 5.87, respectively. Moreover, the deduced PsDR proteins displayed more than 90% similarity to proteins from other Populus species. In response to the avirulent isolate, Sb052, and the virulent isolate, Th053, of MLP, the expression of PsDR genes was rapidly up-regulated in the leaves of P. szechuanica, peaked at 2 or 7 days post-inoculation (dpi), with levels in the incompatible interaction being higher than those in the compatible interaction. Meanwhile, the expression of PsDR genes (except for PsGns) was also differentially up-regulated at 3, 7, or 18 dpi in the petioles of the infected leaves, leaves next to the inoculated leaves, and in the top buds of the infected plants, respectively, compared to that at 0 dpi. These results suggest that these PsDR genes could play distinctive roles in the defense response of poplar against rust infection.

First Report of Powdery Mildew Podoshaera tridactyla on Prunus hypoleuca in China.

Prunus hypoleuca (≡ Maddenia hypoleuca), a native plant in China, grows in the Qinling Mountains that lie at the intersection of several forest regions in north, central, and southwest China. In October 2013, P. hypoleuca suffering from heavy powdery mildew infections was found with approximately 75% of the plants affected. The powdery mildew at first appeared as circular to irregular white patches, which subsequently showed abundant hyphal growth on both sides of leaves, leading to the withering of the leaves. A voucher specimen was maintained in the Mycological Herbarium of Northwest A & F University (Accession No. HMNWAFU-CF 2013166). Hyphal appressoria were nipple-shaped or nearly absent. Conidiophores were cylindrical, measured 83 to 110 × 10 to 12.5 µm, and produced two to five immature conidia in chains with a crenate outline. Foot-cells of conidiophores were straight, cylindrical, and 28 to 62 × 7 to 10 µm. Conidia were hyaline, ellipsoid to ovate, and measured 20 to 32 × 14 to 21 µm (length/width ratio 1.4:1.8). Chasmothecia were scattered or gregarious, depressed globose, and 65 to 112 µm in diameter. Appendages, arising from the upper half of the chasmothecia, usually had two to four dichotomous branches, and were one to three and a half times as long as the chasmothecial diameter. A single ascus in a chasmothecium was subglobose or broadly ellipsoid-ovoid, measured 66 to 86 × 47 to 76 µm and contained six to eight ascospores. The ascospores were ellipsoid-ovoid and 15 to 27 × 12 to 18 µm. The fungus was identified as Podosphaera tridactyla based on its anamorph and teleomorph characteristics (1,2). To confirm the identification, 28S rDNA and the ITS region were amplified. The ITS5/P3 and then PM5/ITS4 primers were used to amplify the ITS region by nested PCR. The primers LSU1/LSU2 were used to amplify the 28S rDNA, and the cloned fragments were sequenced. The 28S rDNA and ITS region sequences were deposited in GenBank (Accession Nos. KJ879240 and KM213121). A GenBank BLAST search of two sequences revealed 99% identity with P. tridactyla infecting Prunus salicina Lindl. in Korea (3). Based on ITS and a 28S rDNA phylogenetic tree, the two sequences retrieved from the Chinese specimen clustered within a strongly supported clade (bootstrap value = 100%) with P. tridactyla (JQ517296 and AB022393, respectively). Cladistic trees were constructed using the neighbor-joining method with the Kimura two-parameter substitution model in MEGA 5.0. Branch robustness was assessed via bootstrap analysis with 1,000 replicates. Phylogenetic analysis data were in agreement with morphological characters (3). To our knowledge, this is the first report of powdery mildew caused by P. tridactyla on P. hypoleuca. While Koch's postulates have not been carried out because of the biotrophic nature of the pathogen, the present report serves as a novel resource in order to improve the understanding of the etiology and epidemiology of the powdery mildew (P. tridactyla) on P. hypoleuca. The occurrence of P. tridactyla, a common powdery mildew on Prunus s. lat., supports recently published results of phylogenetic analyses of the Prunus complex, indicating that Maddenia must be reduced to synonymy with Prunus (4). References: (1) U. Braun. Beih. Nova Hedwigia 89:1, 1987. (2) U. Braun and R. T. A. Cook. Taxonomic Manual of the Erysiphales (Powdery Mildews), CBS Biodiversity Series No. 11. CBS, Utrecht, Netherlands, 2012. (3) S. C. Lee et al. Res. Plant Dis. 18:49, 2012. (4) J. Wen and W. T. Shi. PhytoKeys 17(2):39, 2012.

Identification and characterization of differentially expressed genes during incompatible interaction between the foliar rust Melampsora larici-populina and poplar.

Poplars are extensively cultivated worldwide, and their susceptibility to the foliar rust fungus leads to considerable damages in plantations. To better understand the molecular basis of poplar responses to the foliar rust, suppression subtractive hybridization was used to identify the potential important or novel genes involved in the Populus szechuanica infection by Melampsora larici-populina. A total of 515 expressed sequence tags (ESTs) with high quality were obtained and clustered into 66 contigs and 75 singletons to give a set of 141 uniESTs. The nucleotide Basic Local Alignment Search Tool (BLASTn) program was used to search for homologous sequences of the uniESTs in the GenBank database. Among them, 92.9% showed homology to the poplar genome, and 2% showed similarity to the rust fungus genome. In addition, homology to known genes was analyzed by the BLASTx algorithm, and approximately 50% of the uniESTs were significantly homologous to genes encoding proteins with known functions. Based on a reverse transcription-polymerase chain reaction (PCR) and quantitative PCR approach, five uniESTs were analyzed, and the results showed that the expression level of the thaumatin-like gene was highest at 72 h post-inoculation, and the pathogenesis-related protein 1 gene was highest at 48 h post-inoculation. The information generated in this study provides new clues to aid in the understanding of incompatibility between poplar and the foliar rust.

Partial purification of two forms of choline kinase and separation of choline kinase from sphingosine kinase of rat brain.

Choline kinase of rat brain was purified approximately 200,000 fold using acid precipitation, ammonium sulphate fractionation, Q-Sepharose, Octyl-Sepharose and AH-Sepharose chromatography. The ability of this enzyme to catalyze the phosphorylation of choline, ethanolamine (Etn), monomethylethanolamine (MeEtn), dimethylethanolamine (Me2Etn) and sphingosine was investigated. Choline kinase was separated from sphingosine kinase. The fraction with highly purified choline kinase had four major polypeptides with different molecular masses and possessed activities towards choline, Etn, MeEtn and Me2Etn. Two forms of choline kinase were obtained when the enzymatically active fractions eluted from the Q-Sepharose column were subjected to a horizontal isoelectrofocusing electrophoresis. One form focused around pH 4.7 and is able to phosphorylate choline, Etn, MeEtn and Me2Etn. The other form focused around pH 10 and possessed only choline kinase activity. The latter form of choline kinase did not display classical Michaelis-Menten's mechanism but revealed a positive co-operative pattern for two choline binding sites. This form was purified to apparent homogeneity with a approximate molecular mass of 14.4 kDa.

[Lung cancer with extra pulmonary symptoms as the chief manifestation. A clinical analysis of 47 cases].

In patients with cancer of the lung, extrapulmonary symptoms can be found before or after the tumor is discovered. In this paper, 47 cases of lung cancer presenting extrapulmonary symptoms as their major clinical manifestation are reported. The symptoms might arise from the central nervous system, bones and joints, esophagus, intestine, etc. Some of the symptoms were due to compression or invasion of the cancer to the adjacent organs. Paraneoplastic syndromes, such as ectopic endocrine syndrome and carcinoid syndrome, were also encountered. Keeping these extrapulmonary manifestations in mind, patients suspicious of having lung cancer should be carefully examined and closely followed up until a correct diagnosis is made.

[A long-term follow-up study with endoscope in chronic gastritis].

A 10 to 17 years endoscope follow-up was performed to 138 cases of chronic gastritis. The result showed that 118 cases still proved to be chronic gastritis, and the increase of chronic atrophic gastritis (CAG) in numbers. 15 cases developed into peptic ulcer. 5 cases to carcinoma (4 cases were early carcinoma). The time of cancerization differed from 2 to 12 years. The rate of cancerization of CAG reached 7.46%. The rate of cancerization of intestinal metaplasia (IM) were 8.20%. 13 cases of IM were mucus histo-chemical stained, and five of them contained sulfuric acid mucus, one of the 5 cases cancerized. 3 of 14 cases with atypical hyperplasia (ATP) turned into stomach cancer. We believe that chronic gastritis, especially CAG with ATP and IM, or with sulfuric acid mucus of IM had a high possibility of cancerization with the increase of age, and should be followed up for a long time.

Partial purification and characterization of monomethylethanolamine kinase and dimethylethanolamine kinase activities from rat liver.

Monomethylethanolamine (MEA) kinase and dimethylethanolamine (DEA) kinase activities were purified 950 and 750 fold respectively from rat liver by conventional procedures. Certain properties of the partially purified enzyme preparation suggest that they are different from both choline kinase activity and ethanolamine kinase activity and differ from one another. This is based upon the following observations: 1. The heat stabilities of MEA kinase and DEA kinase activities are significantly different from one another and are different from the stability of choline kinase and ethanolamine kinase activities. 2. K+ in the presence of Mg2+ increases MEA kinase activity by 100% but has no effect on DEA kinase activity. 3. Different Ki values and the types of inhibition by several structurally related amino alcohols were found for MEA kinase and DEA kinase activities. 4. The purification fold of MEA kinase and DEA kinase are different from each other and from that of choline kinase and ethanolamine kinase.

[Distribution of phenotypes of ACP in ten ethnic groups in Yunnan of China].

The distribution of phenotypes of red cell phosphatase (ACP) in ten ethnic groups in Yunnan Province was investigated by starch gel electrophoresis. Three phenotypes A, AB and B were found. The gene frequencies of ACPA and ACPB in Han, Yi, Bai, Dai, Yao, Wa, Hani, Bulang, Jino and Lahu were 0.2067, 0.7933; 0.2406, 0.7594; 0.2341, 0.7659; 0.3750, 0.6250; 0.2300, 0.7700; 0.2727, 0.7273; 0.3594, 0.6406; 0.3036, 0.6964; 0.2381, 0.7619; and 0.4474, 0.5526, respectively. The ACPC gene and other rare genes were not found. The analysis showed that the distribution of phenotypes of ACP1 have some differences in different races and ethnics.

[Subtyping of Pi in human bloodstains by isoelectric focusing].

Subtyping of Pi (alpha 1-AT) in 90 human bloodstains stored under different conditions was carried out by using ULPAGIF. The detectable time of Pi subtypes of all bloodstains kept at room temperature (12-20 degrees C), 4 degrees C and-15 degrees C was 10 days, 4 weeks and 4 weeks respectively. Beyond this time limit, the detectable rate decreased gradually. The minimal detectable quantity, of fresh bloodstains was 4 microliters. The factors influencing the detection of Pi subtypes in bloodstains were discussed.