Prokaryotic expression, subcellular localization and enzymatic activity analysis of DXS gene from Platycodon grandiflorum / 药学学报

1-Deoxy-D-xylulose-5-phosphate synthase (DXS), the first key enzyme in 2-methyl-D-erythritol-4-phosphate (MEP) pathway, catalyzes the condensation of glyceraldehyde-3-phosphate with pyruvate to 1-deoxy-xylose-5-phosphate (DXP). In this study, PgDXS1, PgDXS2, and PgDXS3 genes were cloned from the root of Platycodon grandiflorum (P. grandiflorum). The open reading frame (ORF) of PgDXS1, PgDXS2, and PgDXS3 were 2 160, 2 208, and 2 151 bp in full length, encoding 719, 735, and 716 amino acids, respectively. Homologous alignment results showed a high identity of PgDXSs with DXS in Hevea brasiliensis, Datura stramonium and Stevia rebaudiana. The recombinant expression plasmids of pET-28a-PgDXSs were constructed and transformed into Escherichia coli (E. coli) BL21 (DE3) cells, and the induced proteins were successfully expressed. Subcellular localization results showed that PgDXS1 and PgDXS2 were mainly located in chloroplasts, and PgDXS3 was located in chloroplasts, nucleus and cytoplasm. The expression of three DXS genes in different tissues of two producing areas of P. grandiflorum were assayed via real-time fluorescence quantitative PCR, and the results showed that all of them were highly expressed in leaves of P. grandiflorum from Taihe. Under methyl jasmonate (MeJA) treatment, the expression levels of three PgDXS genes showed a trend of first decreasing and then increasing at different time points (3 - 48 h), and the activity of DXS showed a trend of first increasing and then decreasing in three tissues of P. grandiflorum. This study provides a reference for further elucidating the biological function of PgDXS in terpenoid synthesis pathway in P. grandiflorum.

Complete chloroplast genome sequencing and phylogeny of wild Atractylodes lancea from Yuexi, Anhui province / 中国中药杂志

This study investigated the choroplast genome sequence of wild Atractylodes lancea from Yuexi in Anhui province by high-throughput sequencing, followed by characterization of the genome structure, which laid a foundation for the species identification, analysis of genetic diversity, and resource conservation of A. lancea. To be specific, the total genomic DNA was extracted from the leaves of A. lancea with the improved CTAB method. The chloroplast genome of A. lancea was sequenced by the high-throughput sequencing technology, followed by assembling by metaSPAdes and annotation by CPGAVAS2. Bioiformatics methods were employed for the analysis of simple sequence repeats(SSRs), inverted repeat(IR) border, codon bias, and phylogeny. The results showed that the whole chloroplast genome of A. lancea was 153 178 bp, with an 84 226 bp large single copy(LSC) and a 18 658 bp small single copy(SSC) separated by a pair of IRs(25 147 bp). The genome had the GC content of 37.7% and 124 genes: 87 protein-coding genes, 8 rRNA genes, and 29 tRNA genes. It had 26 287 codons and encoded 20 amino acids. Phylogenetic analysis showed that Atractylodes species clustered into one clade and that A. lancea had close genetic relationship with A. koreana. This study established a method for sequencing the chloroplast genome of A. lancea and enriched the genetic resources of Compositae. The findings are expected to lay a foundation for species identification, analysis of genetic diversity, and resource conservation of A. lancea.

Cloning and expression analysis of superoxide dismutase SmMSD2 gene from Salvia miltiorrhiza / 药学学报

Superoxide dismutase (SOD) is a key enzyme that scavenge superoxide anion free radical (O2·-) in vivo, and plays an important role in plant growth and development and stress. In this study, according to the genome and transcriptome data of Salvia miltiorrhizae, 9 SOD genes were identified and the expression patterns of SOD family genes were further analyzed, including 5 Cu/Zn-SOD, 2 Fe-SOD and 2 Mn-SOD. On the basis of proteomic analysis, combined with transcriptome data, one full-length cDNA of Mn-SOD gene, namely SmMSD2 was cloned from Salvia miltiorrhizae. The results of amino acid sequence alignment and phylogenetic analysis showed that SmMSD2 protein belongs to the manganese superoxide dismutase (Mn-SOD) subfamily, and SmMSD2 protein shares high sequence identity with the Mn-SOD proteins of various plants that all contain a C-terminal conserved metal-binding domain "DVWEHAYY". The prokaryotic expression vector pMAL-c2X-SmMSD2 was constructed and transformed into E. coli BL21 expressing strain, and the target recombinant protein was successfully induced and its enzymatic properties were analyzed. Spatiotemporal expression analysis showed that SmMSD2 gene was expressed in all tissues, indicating that SmMSD2 gene was constitutively expressed at a stable level. Real-time quantitative PCR indicated that drought (15% PEG6000), abscisic acid (ABA) and indole-3-acetic acid (IAA) could induce the expression of SmMSD2 gene, suggesting that SmMSD2 may be involved in the response of Salvia miltiorrhizae to abiotic stress such as drought, as well as the signaling pathways of phytohormone ABA and IAA. These results lay the foundation for further elucidating the involvement of superoxide dismutase in the stress response and accumulation of active components of Salvia miltiorrhiza.

The UGT74L2 of Andrographis paniculata catalyzes phloretin to produce trilobatin and its enzymatic study / 药学学报

The last essential enzyme in the biosynthetic pathway of trilobatin, phloretin-4'-O glycosyltransferase (P4'-OGT), catalyzes the conversion of trilobatin to phloretin in vitro. However, only a few P4'-OGTs have been found in plants. This study used Malus domestica phloretin-4'-O glycosyltransferase (MdPh-4'-OGT) as a query to identify and clone two UDP-glucuronosyltransferase (UGT) genes, designated UGT74L2 and UGT74L3, from the transcriptome of Andrographis paniculata. According to a phylogenetic tree analysis, UGT74L2 and UGT74L3 belonged to the UGT74 family, which has been linked to several activities in other species. The in vitro enzymatic reaction demonstrated that UGT74L2 could particularly catalyze the formation of trilobatin from phloretin, but UGT74L3 had no effects. By using Ni-NTA affinity chromatography to extract the soluble UGT74L2 recombinant protein, the enzymatic kinetics of the activity was investigated using phloretin as the substrate. The results showed that the optimal temperature and pH for UGT74L2 enzymatic reaction were 40 ℃ and 8.0 (Tris-HCl system), respectively. Three metal ions (Ca2+, Mn2+ and Co2+) showed inhibitory effect on the activity of UGT74L2, while Mg2+ could improve the activity of UGT74L2. Other tested metal ions have no significant effect on UGT74L2. The results of enzymatic kinetic parameters that the Km value was 29.84 μmol·L-1, the kcat was 0.02 s-1, and the kcat·Km-1 was 572.6 mol-1·s-1. By homology modeling, molecular docking and mutation experiments, we found that multiple amino acids residues around the substrate binding pocket play quite an important role during catalytic process, In summary, we identified a novel P4'-OGT gene from medicinal plant Andrographis paniculata and provided a new efficient catalyst to synthesize trilobatin. Meanwhile, this study provides a reference for mining new efficient glycosylation modules from plants.

Radiomics based on biparametric MRI for the detection of significant residual prostate cancer after androgen deprivation therapy: using whole-mount histopathology as reference standard / 亚洲男科学杂志(英文版)

We aimed to study radiomics approach based on biparametric magnetic resonance imaging (MRI) for determining significant residual cancer after androgen deprivation therapy (ADT). Ninety-two post-ADT prostate cancer patients underwent MRI before prostatectomy (62 with significant residual disease and 30 with complete response or minimum residual disease [CR/MRD]). Totally, 100 significant residual, 52 CR/MRD lesions, and 70 benign tissues were selected according to pathology. First, 381 radiomics features were extracted from T2-weighted imaging, diffusion-weighted imaging, and apparent diffusion coefficient (ADC) maps. Optimal features were selected using a support vector machine with a recursive feature elimination algorithm (SVM-RFE). Then, ADC values of significant residual, CR/MRD lesions, and benign tissues were compared by one-way analysis of variance. Logistic regression was used to construct models with SVM features to differentiate between each pair of tissues. Third, the efficiencies of ADC value and radiomics models for differentiating the three tissues were assessed by area under receiver operating characteristic curve (AUC). The ADC value (mean ± standard deviation [s.d.]) of significant residual lesions ([1.10 ± 0.02] × 10-3 mm2 s-1) was significantly lower than that of CR/MRD ([1.17 ± 0.02] × 10-3 mm2 s-1), which was significantly lower than that of benign tissues ([1.30 ± 0.02] × 10-3 mm2 s-1; both P < 0.05). The SVM feature models were comparable to ADC value in distinguishing CR/MRD from benign tissue (AUC: 0.766 vs 0.792) and distinguishing residual from benign tissue (AUC: 0.825 vs 0.835) (both P > 0.05), but superior to ADC value in differentiating significant residual from CR/MRD (AUC: 0.748 vs 0.558; P = 0.041). Radiomics approach with biparametric MRI could promote the detection of significant residual prostate cancer after ADT.

Molecular cloning and functional characterization of an isoflavone glucosyltransferase from Pueraria thomsonii / 中国天然药物

Pueraria thomsonii has long been used in traditional Chinese medicine. Isoflavonoids are the principle pharmacologically active components, which are primarily observed as glycosyl-conjugates and accumulate in P. thomsonii roots. However, the molecular mechanisms underlying the glycosylation processes in (iso)flavonoid biosynthesis have not been thoroughly elucidated. In the current study, an O-glucosyltransferase (PtUGT8) was identified in the medicinal plant P. thomsonii from RNA-seq database. Biochemical assays of the recombinant PtUGT8 showed that it was able to glycosylate chalcone (isoliquiritigenin) at the 4-OH position and glycosylate isoflavones (daidzein, formononetin, and genistein) at the 7-OH or 4'-OH position, exhibiting no enzyme activity to flavonones (liquiritigenin and narigenin) in vitro. The identification of PtUGT8 may provide a useful enzyme catalyst for efficient biotransformation of isoflavones and other natural products for food or pharmacological applications.

Cloning and prokaryotic expression analysis of AlCMK from Atractylodes lancea / 药学学报

4-(Cytidine 5′-diphospho)-2-C-methyl-D-erythritol kinase (CMK) was one of the key enzymes in the methylerythritol-4-phosphate (MEP) pathway to generate terpenoids. In this study, based on the transcriptome data of Atractylodes lancea, the sequence of the CMK gene was cloned, named AlCMK (GenBank accession number OM283293). The results showed that AlCMK contains a 1 230 bp open reading frame (ORF) encoding 409 amino acids. The deduced protein had a theoretical molecular weight of 44 752.53 and an isoelectric point of 6.67. Transmembrane structure analysis showed that there was no transmembrane structure, and the secondary structure of AlCMK was predicted to be mainly composed of random coil. Homologous alignment revealed that AlCMK shared high sequence identity with the CMK proteins of Tanacetum cinerariifolium, Osmanthus fragrans, Eucommia ulmoides, Lonicera japonica and Salvia miltiorrhiza. Phylogenetic analysis indicated that AlCMK protein had the higher homology with CMK protein of Compositae. The pET-32a-AlCMK prokaryotic expression vector was constructed and a fusion protein with molecular mass of about 65 kDa was expressed in the E. coli BL21 (DE3). The qRT-PCR was used to analyze the expression pattern of AlCMK gene in different tissues and after MeJA treatment. Meanwhile, the enzyme activity was determined by ELISA kit. The results showed that AlCMK gene was tissue-expressed in different origins and its expression was induced by MeJA, and the results of the enzyme activity assay showed that the AlCMK enzyme activity in different regions was higher in the leaves. The subcellular localization showed that AlCMK was located in the chloroplast. This study provides a reference for further elucidating the biological function of AlCMK gene in terpenoid synthesis pathway in Atractylodes lancea.

Cloning and prokaryotic expression of 3-ketoacyl-CoA thiolase gene AIKAT from Atractylodes lancea / 中国中药杂志

In this study, the gene encoding the key enzyme 3-ketoacyl-CoA thiolase(KAT) in the fatty acid β-oxidation pathway of Atractylodes lancea was cloned. Meanwhile, bioinformatics analysis, prokaryotic expression and gene expression analysis were carried out, which laid a foundation for the study of fatty acid β-oxidation mechanism of A. lancea. The full-length sequence of the gene was cloned by RT-PCR with the specific primers designed according to the sequence information of KAT gene in the transcriptomic data of A. lancea and designated as AIKAT(GenBank accession number MW665111). The results showed that the open reading frame(ORF) of AIKAT was 1 323 bp, encoding 440 amino acid. The deduced protein had a theoretical molecular weight of 46 344.36 and an isoelectric point of 8.92. AIKAT was predicted to be a stable alkaline protein without transmembrane segment. The secondary structure of AIKAT was predicted to be mainly composed of α-helix. The tertiary structure of AIKAT protein was predicted by homology modeling method. Homologous alignment revealed that AIKAT shared high sequence identity with the KAT proteins(AaKAT2, CcKAT2, RgKAT and AtKAT, respectively) of Artemisia annua, Cynara cardunculus var. scolymus, Rehmannia glutinosa and Arabidopsis thaliana. The phylogenetic analysis showed that AIKAT clustered with CcKAT2, confirming the homology of 3-ketoacyl-CoA thiolase genes in Compositae. The prokaryotic expression vector pET-32 a-AIKAT was constructed and transformed into Escherichia coli BL21(DE3) for protein expression. The target protein was successfully expressed as a soluble protein of about 64 kDa. A real-time quantitative PCR analysis was performed to profile the AIKAT expression in different tissues of A. lancea. The results demonstrated that the expression level of AIKAT was the highest in rhizome, followed by that in leaves and stems. In this study, the full-length cDNA of AIKAT was cloned and expressed in E. coli BL21(DE3), and qRT-PCR showed the differential expression of this gene in different tissues, which laid a foundation for further research on the molecular mechanism of fatty acid β-oxidation in A. lancea.

Different development phase of transcriptome analysis from Crataegus pinnatifida Bge. and cloning, structure and function prediction of squalene epoxidase involved in triterpenic acid biosynthesis / 药学学报

italic>Crataegus pinnatifida is a traditional Chinese medicine, which contains organic acids, triterpenoid acids and other active components, has important medicinal and edible value. In order to study the difference of gene expression level in different developmental stages of hawthorn and explore the genes of active ingredient biosynthesis in Crataegus pinnatifida, high-throughput Illumina HiSeq 2000 technology were used to conduct transcriptome sequencing and bioinformatics analysis on Crataegus pinnatifida fruits from the same origin at different developmental stages. 78 496 Unigenes with an average length of 941 nt were obtained by Trinity software. Among them, 58 395 Unigenes can be annotated by NR, NT, Swiss prot, KEGG, COG, GO and other public databases. KEGG pathway analysis showed that 52 Unigenes encoding 15 key enzymes involved in the citric acid cycle. There are 62 Unigenes were involved in the triterpene biosynthesis pathway of Crataegus pinnatifida. Two key enzymes SQE of triterpenoid metabolism pathway in Crataegus pinnatifida were cloned and performed bioinformatic analysis. The results showed that ORF of CpSQE1 and CpSQE2 were 1 594 bp and 1 597 bp, respectively, encoding 530 and 531 amino acids. The molecular weight of proteins was 57.6 kDa and 57.5 kDa. Bioinformatics analysis showed that both CpSQE1 and CpSQE2 proteins have a PLN02985 superfamily conserved domain, belonging to the squalene monooxygenase superfamily. The phylogenetic tree shows that CpSQE1 and CpSQE2 are clustered together with SQE with squalene epoxidase function in other plants. This study provides a research basis for further exploring the key genes in the biosynthesis process of hawthorn active ingredients and analyzing the regulation pathway of its active ingredient biosynthesis.

Evaluating the efficacy and safety of ultrasound-guided percutaneous microwave ablation for the treatment of adrenal metastasis

Context: Microwave ablation (MWA) has been proven as a promising method to treat solid tumors. Aims: This study aims to evaluate the efficacy and safety of ultrasound (US)-guided MWA for treating adrenal metastasis and to explore the factors affecting survival. Settings and Design: This was a retrospective study performed on patients treated at our department. Subjects and Methods: A total of 43 patients with adrenal metastasis (22 hepatocellular carcinoma, eight renal cell carcinoma, five non-small cell lung cancer, four colorectal cancer, three liposarcoma, and one malignant fibrous histiocytoma) were enrolled. All patients were treated at our department at least once. The treatment protocol for each patient, the technique used, and the survival details were recorded. Statistical Analysis Used: Statistical analyses were performed using SPSS 26.0 software. Results: Technical success was achieved in all cases. MWA was a safe technique for treating all types of metastasis. No major complications were observed. The pathology of adrenal lesions was the significant risk factor contributing to overall survival (OS) (P = 0.040). The 1-year and 3-year OS rates for all patients were 0.828 and 0.389, respectively. Conclusions: Percutaneous US-guided MWA is safe and effective in terms of local control and survival of adrenal metastasis

Predictive effects of a combined indicator in patients with hepatocellular carcinoma after thermal ablation

Aim: This study aimed to investigate the predictive power of the combination of Systemic Immune-Inflammation Index (SII) and albumin-bilirubin (ALBI) grade in prognosis outcomes of early-stage hepatocellular carcinoma (HCC) after thermal ablation. Materials and Methods: This retrospective study was reviewed and approved by our institutional review board, and written informed consent was obtained from each patient. According to the Milan criteria, a total of 405 treatment-naïve patients with clinicopathologically confirmed HCC were enrolled who subsequently underwent thermal ablation from 2011 to 2016. The outcomes of overall survival (OS), cancer-specific survival (CSS), and recurrence-free survival (RFS) were statistically analyzed. Results: The median follow-up time of this study was 45.1 months (range, 1.3–83.2 months). After thermal ablation in patients with SII-ALBI Grades 1, 2, and 3, the cumulative 5-year OS rates were 81.7%, 63.2%, and 26.9%; the 5-year CSS rates were 82.4%, 67.5%, and 26.9%; and the 5-year RFS rates were 49.3%, 44.6%, and 25.3%, respectively (all P < 0.001). On multivariate Cox regression analyses, SII-ALBI was independently associated with the three outcomes after adjustment for various confounders (all P < 0.05). In addition, SII-ALBI played a predictive role in OS, CSS, and RFS for patients with negative alpha-fetoprotein (AFP) (P < 0.05). Compared with SII and ALBI, the AUCs for the prediction of OS and CSS using SII-ALBI were superior to single indicator (bothP < 0.05). Conclusion: Elevated preablation SII-ALBI is associated with shorter OS, CSS, and RFS in patients with early-stage HCC. Our indicator showed the potential to be a supplement tool for patients with negative AFP during follow-up

Transarterial chemoembolization combined with microwave ablation versus microwave ablation only for Barcelona clinic liver cancer Stage B hepatocellular carcinoma: A propensity score matching study

Purpose: We aimed to compare the outcomes of microwave ablation (MWA) alone with those of transarterial chemoembolization combined with MWA (TACE-MWA) for Barcelona clinic liver cancer (BCLC) Stage B hepatocellular carcinoma (HCC) and to identify the prognostic factors associated with the two treatments. Materials and Methods: This retrospective study was conducted in 150 BCLC Stage B HCC patients from April 2006 to November 2017. Of these, 88 patients were treated with MWA alone while 62 with TACE-MWA. Propensity score matching (PSM) was conducted to adjust for imbalances in clinical parameters. Procedure-related complications, local tumor progression (LTP), recurrence-free survival (RFS), and overall survival (OS) were analyzed. Results: Before PSM, the maximal tumor diameters were 6.0 ± 1.0 cm and 6.7 ± 1.3 cm in the TACE-MWA and MWA groups, respectively, with a significant difference (P = 0.002); a significant difference was also detected in α-fetoprotein level (P = 0.013). After PSM, no difference was found in the two parameters (P = 0.067, 0.470). Before and after PSM, no difference was detected in the procedure-related complications (P = 0.803 vs. 1.000, P = 1.000 vs. 1.000), RFS (P = 0.786 vs. 0.689), and OS (P = 0.684 vs. 0.929). Tumor size and α-fetoprotein level were independent influencing factors for OS before and after PSM (P = 0.009, 0.023), while tumor size (D > 7) was an independent risk factor for poor OS (P = 0.011). Tumor number was an independent risk factor for RFS before and after PSM (P = 0.007 vs. P = 0.008). A significant difference was detected in LTP between the two groups with single tumor before and after PSM (P = 0.059 vs. P = 0.006). Conclusions: The MWA alone group had RFS and OS comparable to that of the TACE-MWA group. TACE-MWA was effective in controlling LTP in patients with a single tumor

Microwave ablation versus other interventions for hepatocellular carcinoma: A systematic review and meta-analysis

We aim to evaluate the efficacy and safety of microwave ablation (MWA) versus other treatment modalities for hepatocellular carcinoma (HCC). This study was registered in Prospero (registration number CRD42017057046). A complete electronic search was conducted for studies on MWA versus other interventions for HCC using PubMed, EMBASE, Cochrane Library databases, and ISI Web of Science. Randomized and non-randomized clinical trials were included. Data on technical efficacy, local tumor progression (LTP), overall survival (OS), progression-free survival (PFS), and major complications were extracted from included studies and combined to be analyzed via random effects models. OS was set as the primary outcome measure. Fifteen clinical studies were identified. When comparing MWA with radiofrequency ablation (RFA), no significant difference was found in 3-year OS rates (odds ratio [OR] 0.94, 95% confidence interval [CI] 0.66–1.34, P = 0.74), 5-year OS rates (OR 0.83, 95% CI 0.58–1.18, P = 0.29), 3-year PFS rates (OR 1.05, 95% CI 0.77–1.43, P = 0.74), 1-year LTP rate (OR 1.28, 95% CI 0.52–3.18,P = 0.59), technical efficacy rate (OR 1. 35, 95% CI 0. 85–2.15, P = 0.20), and major complication rate (OR 1.04, 95% CI 0.56–1.93, P = 0.90). When comparing MWA with hepatic resection, the 3-year OS rate was not significantly different (OR 0.89, 95% CI 0.59–1.35, P = 0.59). Compared with RFA and hepatic resection, MWA showed similar safety and efficacy for HCC, especially in OS rate and PFS. However, high-quality clinical trials are needed to validate the superiority of MWA

US-guided percutanous microwave ablation for early-stage hepatocellular carcinoma in elderly patients is as effective as in younger patients: A 10-year experience

Objective: To compare the overall survival (OS), disease-free survival (DFS) and liver-cancer-specific survival (LCSS) of elderly (≥65 years) and younger patients (< 65 years) with early-stage hepatocellular carcinoma (HCC) using ultrasound-guided percutaneous microwave ablation (US-PMMA). Materials and Methods: From January 2002 to December 2017, 510 elderly and 1053 younger patients were diagnosed with early-stage HCC according to the Milan criteria. All of these patients were treatment-naïve to US-PMMA. Baseline characteristics were collected to identify any risk factors to determine the survival outcomes. OS, DFS, and LCSS probabilities were calculated with the Kaplan-Meier method and compared using the Log-rank test. Results: Complete ablation was achieved in all patients. Elderly patients were more likely to be, hepatitis C virus infection, comorbidities, cirrhosis, larger tumors, poor liver functional reservation, more ablation points, longer ablation time, longer hospital stays, and higher hospitalization costs (P < 0.05). Over the follow-up period (12–156 months), no significant differences were detected in OS, DFS, and LCSS between the two groups ( P = 0.092, 0.318, and 0.183). r-GT, ALB and ablation session were significant factors for OS, r-GT and ALB for LCSS, and cirrhosis, tumor number, AFP and ablation points for RFS in the multivariate analysis, respectively. No treatment-related deaths occurred in the two groups. Any complications were treated as appropriate. Conclusions: Although advanced age and comorbidities are intrinsic factors in elderly HCC patients, similar survival outcomes were obtained in elderly and younger HCC patients treated by US-PMWA, despite elderly patients having more comorbidities

Ultrasound-guided percutaneous microwave ablation assisted by a three-dimensional visualization preoperative treatment planning system for larger adrenal metastasis (D ≥ 4 cm): Preliminary results

Objective: The objective of this study is to assess the clinical effect and safety of ultrasound-guided percutaneous microwave ablation (US-PMWA) assisted by three-dimensional (3D) visualization operative treatment planning system in larger adrenal metastasis (LAM) (D ≥ 4 cm). Materials and Methods: From Dec 2011 to Dec 2017, 12 consecutive LAM patients with pathologically proven with a mean diameter of 5.2±1.3cm (range 4.1-7.6) were treated. Artificial ascites and thermal monitoring system as ancillary technique were used. The patients were followed up with imagings and complications were recorded. Results: The median follow-up period was 31 months (ranged 6–52 m). All LAM achieved completely ablation according to the 3D planning preoperation. Complete ablation was achieved in 10 (10/12, 83.3%) patients by one session and 2 patients (2/12, 16.7%) by two sessions. Recurrence was detected at the treated site in 3 patients (3/12, 25.0%) at 5, 9, and 13 months after ablation and received another ablation. Progression of metastasis disease at extra-adrenal sites occurred in 9 patients (9/12, 75%). Seven (7/12, 58.3%) patients died during the follow-up period. Therefore, the 1-, 2-, and 3-year local tumor control rates were 83.3%, 75.0%, and 75.0%, and 1-, 2-, 3- and 4-year overall survival rates were 91.7%, 75.0%, 50.0%, and 41.7%, respectively. No severe complications related to ablation occurred, except 3 (3/12, 25%) patients developed hypertension during ablation. Conclusions: US-PMWA assisted by 3D visualization preoperative treatment planning system maybe a safe and efficient therapy for LAM, which could promote ablation precision, improve the clinical outcomes

Local tumor control of thoracoabdominal wall seeding tumor from hepatocellular carcinoma with ultrasound-guided interventional treatment: A summarized study

Objective: The aim of the study is to evaluate the effectiveness of ultrasound (US)-guided interventional treatments in local tumor control (LTC) for thoracoabdominal wall seeding tumor (TAWST) from hepatocellular carcinoma (HCC), and explore the survival outcomes of the patients underwent surgical resection (SR) and microwave ablation (MWA) for intrahepatic tumor. Materials and Methods: A total of 40 patients with 47 TAWST from HCC were recruited from April 2007 to May 2018. LTC was evaluated by contrast-enchanced image. Long-time survival outcomes were compared. Overall survival (OS), disease-free survival (DFS), and local seeding progress-free survival (LSPFS) were analyzed. Results: One-year LTC rate was 65.2% in all patients; 72%, 0%, 50%, and 0% in the patients who underwent MWA, high-intensity focused US, iodine-125 (125I) brachytherapy and MWA combined with 125I brachytherapy, respectively. The 3-year OS, DFS rates and LSPFS rates were 51.8% and 28.6%, 12.0% and 23.8%, and 10.0% and 10.0% after MWA and SR, respectively. Univariate analysis results showed that age (P = 0.049), Karnofsky performance scale (KPS) (P = 0.002), and chemoradiation (P = 0.032); and multivariate analysis results showed that age (P = 0.045) and KPS (P = 0.010) might be predictors for LCT. While univariate analysis results showed that KPS (P = 0.032), intrahepatic tumor size (P = 0.006), chemoradiation (P = 0.003), preoperative alpha-fetoprotein level (P = 0.007), metastasis (P = 0.049), and albumin-bilirubin grade (P = 0.002), and multivariate analysis results showed that comorbidities (P = 0.004), KPS (P = 0.007), and metastasis (P = 0.009) might be predictors for OS. The pain degree of patients was improved obviously after treatments. Conclusions: US-guided interventional treatments were an effective option in LTC for TAWST from HCC, and MWA could achieve comparable long-time survival outcomes with SR for HCC patients with TAWST

Identification of Corydalis yanhusuo,C. turtschaninovii,C. decumbens by allele-specific PCR / 中国中药杂志

To establish a DNA molecular markers method for identification of Corydalis yanhusuo,C. turtschaninovii and C. decumbens,the mat K,trn G and psb A-trn H sequences of 56 samples from 14 species of C. yanhusuo,C. turtschaninovii,C. decumbens and their related species were obtained by sequencing. The SNP loci were obtained by Bio Edit 7. 2. 2 software. The primers for AS-PCR identification were designed based on the mutation sites,and the conditions of PCR were optimized to identify C. yanhusuo,C. turtschaninovii,and C. decumbens according to the specific bands. The results showed that the amount of template( 0. 6-1 200 ng)and annealing temperature( 42-60 ℃) had little influence on the amplification results,and the number of cycles had much influence on the amplification results. When the number of cycles was 20,the specific bands of 297 bp( mat K),353 bp( trn G) and 544 bp( mat K) were amplified from C. yanhusuo,C. turtschaninovii and C. decumbens,respectively. The method established in this study had a minimum detection limit of 6 ng for C. yanhusuo,60 ng for C. decumbens and less than 0. 6 ng for C. turtschaninovii. Thus,the allelespecific PCR method established in the research can specifically identify C. yanhusuo,C. turtschaninovii,and C. decumbens.

Species and medical history of "Xishuang" medicinal materials / 中国中药杂志

"Xishuang" is a special phenomenon that chemical composition of medicinal materials crystallize on the surface exposed to air for a long time. We summarized Herbal textual research of "Xishuang" phenomenon of six herbs, such as Schisandrae Chinensis Fructus, Moutan Cortex, Atractylodis Rhizoma, Magnoliae Officinalis Cortex, dried persimmon frost and watermelon frost. From historical perspective, cream of Schisandrae Chinensis Fructus was firstly discovered in Lei Gong's Moxibustion Theory. Thereafter, dried persimmon frost was found in Song Dynasty, which was named "white persimmon" in Ben Cao Tu Jing and had become an independent medicine in Compendium of Materia Medica. Then, watermelon frost was found in Yang Yi Da Quan of the Qing Dynasty, and Moutan Cortex's "sand star" was recorded in Zeng Ding Wei Yao Tiao Bian of the Republic of China. After that, "Xishuang" phenomenon of Atractylodis Rhizomaand Magnoliae Officinalis Cortex were reported in 1950s and 1960s in succession. The pattern of "Xishuang" is divided into different type, natural "Xishuang" includes Schisandrae Chinensis Fructus, Moutan Cortex, Atractylodis Rhizoma and Magnoliae Officinalis Cortex, artificial "Xishuang" includes watermelon frost, and dried persimmon frost formed crystals by using artificial intervention. The above 6 kinds of herbs have different crystal structure and chemical composition. Therefore, according to traditional identification experience, "Xishuang" phenomenon is related to varieties and quality of medicinal herbs. These research provide herbalism basis for the modern study of "Xishuang" medicinal materials.