Effect of Rhizophagus intraradices on growth of Salvia miltiorrhiza / 中国中药杂志

The study aimed to explore the effects of inoculation of Rhizophagus intraradices on the biomass, effective component content, and endogenous hormone content of Salvia miltiorrhiza through pot experiments. The number of leaves, plant height, dry weight of aboveground and underground parts, branch number, root number, root length, root diameter, and other biomass were mea-sured by weighing and counting methods. The content of salvianolic acid B, caffeic acid, rosmarinic acid, tanshinone Ⅰ, tanshinone Ⅱ_A, cryptotanshinone, and other effective components was determined by ultra-high performance liquid chromatography. The content of ABA and GA_3 was determined by triple quadrupole mass spectrometry. The correlations between biomass and effective components and between effective components and plant hormones ABA and GA_3 were analyzed. The results showed that R. intraradices significan-tly increased the aboveground dry weight, leaf number, and root number of S. miltiorrhiza by 0.24-0.65 times, respectively. The content of salvianolic acid B and rosmarinic acid in the aboveground part and the content of salvianolic acid B, caffeic acid, rosmarinic acid, tanshinone Ⅰ, and tanshinone Ⅱ_A in the underground part were significantly increased by 0.44-1.78 times, respectively. R. intraradices infection significantly increased the GA_3/ABA values of aboveground and underground parts by 3.82 and 76.47 times, respectively. The correlation analysis showed that caffeic acid, the effective component of the aboveground part, was significantly positively correlated with plant height, tanshinone Ⅱ_A, the effective component of the underground part, was significantly positively correlated with biomass root number, cryptotanshinone, and dry weight, while rosmarinic acid was significantly negatively correlated with dry weight. There were significant positive correlations between cryptotanshinone and ABA, tanshinone Ⅱ_A and ABA and GA_3, and caffeic acid and GA_3. In conclusion, R. intraradices can promote the accumulation of biomass and secondary metabolites of S. miltiorrhiza and regulate the balance between plant hormones ABA and GA_3, thereby promoting the growth of S. miltiorrhiza.

A novel homozygous frameshift variant in DNAH8 causes multiple morphological abnormalities of the sperm flagella in a consanguineous Pakistani family / 亚洲男科学杂志(英文版)

Multiple morphological abnormalities of the sperm flagella (MMAF) is a severe form of asthenozoospermia categorized by immotile spermatozoa with abnormal flagella in ejaculate. Whole-exome sequencing (WES) is used to detect pathogenic variants in patients with MMAF. In this study, a novel homozygous frameshift variant (c.6158_6159insT) in dynein axonemal heavy chain 8 (DNAH8) from two infertile brothers with MMAF in a consanguineous Pakistani family was identified by WES. Reverse transcription-polymerase chain reaction (RT-PCR) confirmed DNAH8 mRNA decay in these patients with the DNAH8 mutation. Hematoxylin-eosin staining and transmission electron microscopy revealed highly divergent morphology and ultrastructure of sperm flagella in these patients. Furthermore, an immunofluorescence assay showed the absence of DNAH8 and a reduction in its associated protein DNAH17 in the patients' spermatozoa. Collectively, our study expands the phenotypic spectrum of patients with DNAH8-related MMAF worldwide.

A recurrent homozygous missense mutation in CCDC103 causes asthenoteratozoospermia due to disorganized dynein arms / 亚洲男科学杂志(英文版)

Asthenoteratozoospermia is one of the most severe types of qualitative sperm defects. Most cases are due to mutations in genes encoding the components of sperm flagella, which have an ultrastructure similar to that of motile cilia. Coiled-coil domain containing 103 (CCDC103) is an outer dynein arm assembly factor, and pathogenic variants of CCDC103 cause primary ciliary dyskinesia (PCD). However, whether CCDC103 pathogenic variants cause severe asthenoteratozoospermia has yet to be determined. Whole-exome sequencing (WES) was performed for two individuals with nonsyndromic asthenoteratozoospermia in a consanguineous family. A homozygous CCDC103 variant segregating recessively with an infertility phenotype was identified (ENST00000035776.2, c.461A>C, p.His154Pro). CCDC103 p.His154Pro was previously reported as a high prevalence mutation causing PCD, though the reproductive phenotype of these PCD individuals is unknown. Transmission electron microscopy (TEM) of affected individuals' spermatozoa showed that the mid-piece was severely damaged with disorganized dynein arms, similar to the abnormal ultrastructure of respiratory ciliary of PCD individuals with the same mutation. Thus, our findings expand the phenotype spectrum of CCDC103 p.His154Pro as a novel pathogenic gene for nonsyndromic asthenospermia.

Construction and validation of lncRNA prognostic model for bladder cancer / 中华检验医学杂志

Objective:This study aims to construct a prognostic model of bladder cancer (BLCA) based on lncRNA.Methods:BLCA lncRNA expression data and clinical information were downloaded from TCGA. Univariate Cox regression was used to evaluate the correlation between the expression level of each lncRNA and overall survival (OS), and the lncRNAs with a corrected P-value<0.01 were selected as candidate predictors. In the training queue, the prediction model is constructed by methods such as least absolute shrinkage and selection operator, and multi-factor stepwise Cox regression, and verified in the verification queue at the same time.. Evaluation the area under the curve of time-dependent receiver operating characteristic (tROC) and Harrel C index. According to the median risk score of the prediction model, patients were divided into high-risk group and low-risk group and the differences in clinicopathological characteristics between the two groups were compared by t-test or chi-square test. Results:Establish a BLCA prognostic model based on 13 lncRNAs, of which LINC01465, ARHGAP5-AS1, ZFHX4-AS1, MAFG-AS1 are prognostic risk factors (β regression coefficients are 0.32, 0.16, 0.06, 0.20, respectively, all>0), and the rest are protection factors (β regression coefficients are all<0); the prediction model of the overall survival in the first year, the third year, and the fifth year in the complete cohort has an area under the tROC curve of 0.79, 0.82, and 0.80 respectively, and the Harrell C index is 0.74. Its predictive ability is better than the previously published BLCA prognostic model based on lncRNA. Adjusting for confounding factors including age and tumor stage found that the risk score of this model was an independent poor prognostic factor for overall survival in BLCA patients (hazard ratio 4.05; P<0.001). Comparison of clinicopathological characteristics of patients in the high-risk and low-risk groups showed that in the high-risk group, there were more old patints (70.0 vs. 66.1, P<0.001), more non-papillary patients (74.2% vs. 61.2, P=0.005), more high-stage patients (37.6% vs. 28.0%, P<0.001 for stage Ⅳ patients), and more high-grade tumors (98.0% vs. 92.0%, P=0.005). Conclusion:In this study, a prognostic model of bladder cancer based on 13 lncRNAs was constructed. This model has good predictive ability and can provide value for clinical decision-making and patient consultation.

Effect of different fungicides on efficiency of mycorrhizal symbiosis on Salvia miltiorrhiza / 中国中药杂志

Arbuscular mycorrhizal fungi provided is beneficial to Salvia miltiorrhiza for increasing yield, promoting the accumulation of active ingredients, and alleviating S. miltiorrhiza disease etc. However, the application of fungicides will affect the benefit of arbuscular mycorrhizal fungi and there is little research about it. This article study the effect of four different fungicides: carbendazim, polyoxin, methyl mopazine, and mancozeb on mycorrhiza benefit to S. miltiorrhiza by the infection intensity of arbuscular mycorrhizal fungi, the growth of S. miltiorrhiza, and the content of active ingredients. RESULTS:: showed that different fungicides had different effects. The application of mancozeb had the strongest inhibitory effect on the mycorrhizal benefit to S. miltiorrhiza. Mancozeb significantly reduced the mycorrhizal colonization and the beneficial effect of arbuscular mycorrhizal fungi on the growth and the accumulation of active components of S. miltiorrhiza. The application of polyoxin had no significant effect on mycorrhizal colonization. Instead, it had a synergistic effect with the mycorrhizal benefit to promoting the growth and accumulation of rosmarinic acid of S. miltiorrhiza. The inhibitory strengths of four fungicides are: mancozeb>thiophanate methyl, carbendazim>polyoxin. Therefore, we recommend applying biological fungicides polyoxin and avoid applying chemical fungicides mancozeb for disease control during mycorrhizal cultivation of S. miltiorrhiza.

Novel biallelic loss-of-function mutations in / 亚洲男科学杂志(英文版)

Multiple morphological abnormalities of the sperm flagella (MMAF) is a specific type of asthenoteratozoospermia, presenting with multiple morphological anomalies in spermatozoa, such as absent, bent, coiled, short, or irregular caliber flagella. Previous genetic studies revealed pathogenic mutations in genes encoding cilia and flagella-associated proteins (CFAPs; e.g., CFAP43, CFAP44, CFAP65, CFAP69, CFAP70, and CFAP251) responsible for the MMAF phenotype in infertile men from different ethnic groups. However, none of them have been identified in infertile Pakistani males with MMAF. In the current study, two Pakistani families with MMAF patients were recruited. Whole-exome sequencing (WES) of patients and their parents was performed. WES analysis reflected novel biallelic loss-of-function mutations in CFAP43 in both families (Family 1: ENST00000357060.3, p.Arg300Lysfs*22 and p.Thr526Serfs*43 in a compound heterozygous state; Family 2: ENST00000357060.3, p.Thr526Serfs*43 in a homozygous state). Sanger sequencing further confirmed that these mutations were segregated recessively in the families with the MMAF phenotype. Semiquantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) was carried out to detect the effect of the mutation on mRNA of the affected gene. Previous research demonstrated that biallelic loss-of-function mutations in CFAP43 accounted for the majority of all CFAP43-mutant MMAF patients. To the best of our knowledge, this is the first study to report CFAP43 biallelic loss-of-function mutations in a Pakistani population with the MMAF phenotype. This study will help researchers and clinicians to understand the genetic etiology of MMAF better.

Application of traditional Chinese medicine(TCM) traceability system based on TCM quality characteristics and HACCP system / 中国中药杂志

It is crucial to establish a complete set of traditional Chinese medicine(TCM) quality traceability management process system, in order to stabilize the pricing order of TCM market and reconstruct the transmission path of TCM quality signals. In this study, we reviewed the mature experience of food and drug supervision at home and abroad, analyzed the quality characteristics of TCM, and put forward that the quality control of TCM products can learn from the hazard analysis and critical control point(HACCP) system in food safety quality control. This study points out that the HACCP system provides not only technical guidance for the traceability management of TCM, but also ideas for improving the quality of TCM products and the safety risk control of TCM. The application of the HACCP system in TCM quality control can help establish an international dialogue platform for TCM and help realize the modernization and internationalization of TCM industry.

Development of ultrasound in diagnosis of bone fractures / 中国医学影像技术

Ultrasound can supply information of the position, size, displacement of fracture end and edema or hematoma of surrounding tissue through multi-directional scanning, which has been used in diagnosis of bone fractures, especially in some occult fractures in ribs, metacarpals, cartilages, nasal bones and so on. The application and development of ultrasound in diagnosis of bone fractures were reviewed in this article.

Research and application progress of Lepidium meyenii（maca） / 中国中药杂志

Lepidium meyenii（maca）was a herbaceous plant of the family Cruciferae. It is native to the andes region of South America where the local people had been growing and consuming maca for centuries. The unique chemical composition and physiological function of maca were widely concerned worldwide. It was introduced to China in 2002, and were cultivated successfully in Yunnan, Tibet, Sichuan, Jilin and other places with a certain size. Maca contained not only rich nutrition such as protein, vitamin and mineral matter, but also lots of secondary metabolites as maca alkaloids, glucosinolates, volatile oils, sterols polyphenols and macaenes. Numerous studies suggested that maca may serve effects in resisting oxidation, fatigue resistance, raising fertility, regulating endocrine, enhancing immunity, tumour suppression, treating osteoporosis, regulate blood sugar and protection of nervous system. Maca was approved by the Ministry of Health as a new resource food in 2011, and its related products include food, health foods, cosmetics, etc. Certain exploratory researches were carried to take better advantage of maca's medicinal value. This paper briefly reviewed the research and application progress of maca in recent years from the aspects of botany, chemical composition, function, resources situation and related products development, which was supposed to provide reference for scientific research and utilization of maca.

Effect of soil pH on efficiency of mycorrhizal symbiosis on Salvia miltiorrhiza / 中国中药杂志

By comparing the effects of soil pH on the efficiency of mycorrhizal symbiosis on Salvia miltiorrhiza， the study is aimed to provide guidance for the use of mycorrhiza in the cultivation of S. miltiorrhiza. In this experiment, the inoculant treated and the non-inoculant treated control were grown in different soil pH. The data was collected after 60 days of cultivation including rate of mycorrhizal infection, biomass, and three chemical constituents with known medicinal action. The results showed that Glomus versiforme was more apt to infect S. miltiorrhiza (F>94.00%; M>69.45%; m>73.66%) and promote the growth of S. miltiorrhiza under pH 5-9 soil. The mycorrhizal contribution to the growth of S. miltiorrhiza was the highest when grown in pH 8 soil. Plants grown with mycorrhiza in pH 8 soil had above-ground biomass more than 2 times and root biomass more than 5 times. The uninoculated plants grew better under acidic and neutral conditions, but the inoculated plants grew better under alkaline (pH 8) conditions. This result showed mycorrhiza can play a role in the adaptability of S. miltiorrhiza to the environment. Inoculation of mycorrhiza significantly increased the accumulation of rosmarinic acid, salvianolic acid B, and dihydrotanshinone by 6.59,5.03 and 2.20-folds. Based on our results alkaline soil (pH 8) is most suitable for the cultivation of S. miltiorrhiza by inoculation with the mycorrhiza G. versiforme.

Study on characteristics and microscopic identification of fruits of Notopterygium franchetii and N. forrestii / 中国中药杂志

Rhizoma et Radix Notopterygii is a rare and endangered Chinese medicine. In the collection of Notopterygium franchetii fruits, we collected a sample of N. forrestii , which is a spurious breed. Fruits of N. franchetii and N. forrestii are very similar in morphology and can be easily confused. Until now the morphological identification of the fruits of Notopterygium has not been reported. To provide a scientific basis for the identification of N. franchetii and N. forrestii fruits, the morphology and microscopic identification were studied in this paper. In this study, stereomicroscope and paraffin sections were used to compare the morphological characteristics and microscopic characteristics of these two fruits. Our results showed that these two fruits were different in size, surface texture and the number of vertical edges on the back. These traits can be used as diagnostic characteristic of these two fruits. The difference between the number of tubing and the endosperm cell contents can be used as microscopic identification features. The above discriminative characteristics can distinguish the two fruits and provide scientific basis for the identification and germplasm evaluation of Notopterygium fruits.

Analysis of several key problems of good agricultural practice (GAP) of Chinese materia medica / 中国中药杂志

This paper reviewed the historical background of the GAP, analyzed the development experience of five Ps (GMP, GLP, GSP, GCP, GAP), analyzed the GAP based on economic theories, and pointed out that the core problem of GAP is ignoring economic laws. Once the GAP, is a process of certification, but neither the GAP announcement could be transformed into signal transmission quality in the product market, nor consumers could recognize the difference between GAP and non-GAP herbs in the terminal market, so manufacturers lack motivation for GAP certification. In this paper, we pointed out, that the GAP certification system should be redesigned under the guidance of economics, third party certification body, supervised by drug administration organization, to certificate GAP as high quality standards, to improve signal transduction mechanism of GAP certification, and to integrate GAP with the market.

Effects of arbuscular mycorrhizal fungi on endogenous hormones in Salvia miltiorrhiza / 中国中药杂志

To investigate the dynamic changes of endogenous hormones contents in Salvia miltiorrhiza after the inoculation of Glomus versiforme(GV).The contents of endogenous hormones of ABA, ZR, GA, IAA and MeJA by ELISA were measured. Infection rata of GV reached plateau of 90% at the 90th d of inoculation; fresh weight of overground part and leaf number were significantly higher in GV group, with 2.7 and 1.96 fold than that of control; contents of all endogenous hormones apart from ABA in over- and under-ground part were markedly lower (P<0.05) in GV group at the 75th and 90th d, respectively, with 63% to 75% and 45% to 81% of that in control, and were significantly higher (P<0.05) in both over- and under-ground part in GV group at the 105th d, with 1.4 to 1.7 fold higher than that of control; content of ABA in underground part increased significantly at 60th, 75th and 105th d. Arbuscular mycorrhizal fungi could promote the growth of S. miltiorrhiza, and affect the dynamic changes of endogenous hormones contents in different infection periods.

Effect of herb residue on growth and active ingredient content of licorice / 中国中药杂志

Traditional Chinese medicine industry product a lot of herb residue. Herb residue was treated as household waste. This treatment leads to environmental pollution and resource waste. For this case, we study the effect of different herb residues on the growth and active ingredient content of Licorice by random control experiment. Our results showed that the effects of different herb residues were difference. Atractylodes macrocephala residue and Forsythia suspense residue had the stronger effect and the effect of A. macrocephala residue was inferior to the effect of F. suspense residue. A.macrocephala residue significantly improved the shoot biomass banch number, leaf number, root diameter and root biomass by 0.53-1.81 fold. A. macrocephala residue also increased the glycyrrhizic acid content of root by 1.54 fold. F. suspense residue significantly improved the shoot biomass,branch number, root diameter and root biomass by 0.43-1.13 fold. Four kind herb residues all improved the shoot biomass by 0.43-1.81fold. So, the authors recommand to considered that we can apply A. macrocephala residue and F. suspense residue in Licorice cultivation.

Effect of herb residues compost on growth and phenols of Perilla frutescens / 中国中药杂志

Herb residue is post-decoction material that can be used as organic fertilizer. Unfortunately, it is currently disposed of as solid waste. This method of disposal is a waste of this resource and a source of environmental pollution. For this case,we studied effects of six different herb residues compost on growth and phenols of Perilla frutescens by pot experiment. Our results show that all six herb residues can improve the growth of P. frutescens. The order of their efficiencies was as follows： Salviae Miltiorrhizae Radix residue>Hordei Fructus germinates residue>Forsythia fructus residue>Atractylodis Macrocephalae Rhizome residue＞Sophorae Flavescentis Redix residue and Moutan cortex residue. Effects of Sophorae Flavescentis Radix residue and Moutan Cortex residue weren't significantly different from CK. Six herb residue all improve root system architecture and leaf area. To phenols of P. frutescens, six herb residues all increased the rosmarinic acid and caffeic acid content of root, and accumulation of four phenols. All the analysis showed herb residues compost can improve the growth and four phenols accumulation of P. frutescens, and Salviae Miltiorrhizae Radix residue had the most pronounced effect on P. frutescens.

Effect of five fungicides on growth of Glycyrrhiza uralensis and efficiency of mycorrhizal symbiosis / 中国中药杂志

In order to obtain the fungicides with minimal impact on efficiency of mycorrhizal symbiosis, the effect of five fungicides including polyoxins, jinggangmycins, thiophanate methylate, chlorothalonil and carbendazim on the growth of medicinal plant and efficiency of mycorrhizal symbiosis were studied. Pot cultured Glycyrrhiza uralensis was treated with different fungicides with the concentration that commonly used in the field. 60 d after treated with fungicides, infection rate, infection density, biomass indexes, photosyn- thetic index and the content of active component were measured. Experimental results showed that carbendazim had the strongest inhibition on mycorrhizal symbiosis effect. Carbendazim significantly inhibited the mycorrhizal infection rate, significantly suppressed the actual photosynthetic efficiency of G. uralensis and the most indicators of biomass. Polyoxins showed the lowest inhibiting affection. Polyoxins had no significant effect on mycorrhizal infection rate, the actual photosynthetic efficiency of G. uralensis and the most indicators of biomass. The other three fungicides also had an inhibitory effect on efficiency of mycorrhizal symbiosis, and the inhibition degrees were all between polyoxins's and carbendazim's. The author considered that fungicide's inhibition degree on mycorrhizal effect might be related with the species of fungicides, so the author suggested that the farmer should try to choose bio-fungicides like polyoxins.

Hyperspectral characteristics of Carthamus tinctorius in Xinjiang region / 中国中药杂志

Investigation of traditional Chinese medicine resources is the most important issue of the protection and use of traditional Chinese medicine resources. Real-time monitoring of medicinal plant species and coverage of an area are of great significance to the sustainable development of the medicinal plant species diversity and ecological environment. Flower has unique spectral characteristics. Comparing the vegetative stage through the flowering stage it is easier to identify species. The flowering stage is a critical period for identifying species with remote sensing. Carthamus tinctorius as a traditional Chinese medicine resources in XinJiang region, attracted widespread attention in recent years. In this paper, the hyperspectral data of canopy and other vegetation canopy was measured in 2011. The spectral curve was analyzed, the result indicated that C. tinctorius canopy and the canopy after picking showed absorption peak near 770 nm, the first derivative of red edge spectra and invert-Gaussian model were analyzed, the result indicated that there was significant difference between C. tinctorius canopy and other vegetation canopy. The proposed method is designed to provide initial theoretical foundation for growth condition and physiological parameters of C. tinctorius, and make theoretical groundwork for the distribution and elaborate monitoring of C. tinctorius in future.

Plasma membrane calcium ATPase 4b inhibits nitric oxide generation through calcium-induced dynamic interaction with neuronal nitric oxide synthase

The activation and deactivation of Ca(2+)- and calmodulindependent neuronal nitric oxide synthase (nNOS) in the central nervous system must be tightly controlled to prevent excessive nitric oxide (NO) generation. Considering plasma membrane calcium ATPase (PMCA) is a key deactivator of nNOS, the present investigation aims to determine the key events involved in nNOS deactivation of by PMCA in living cells to maintain its cellular context. Using time-resolved Förster resonance energy transfer (FRET), we determined the occurrence of Ca(2+)-induced protein-protein interactions between plasma membrane calcium ATPase 4b (PMCA4b) and nNOS in living cells. PMCA activation significantly decreased the intracellular Ca(2+) concentrations ([Ca(2+)]i), which deactivates nNOS and slowdowns NO synthesis. Under the basal [Ca(2+)]i caused by PMCA activation, no protein-protein interactions were observed between PMCA4b and nNOS. Furthermore, both the PDZ domain of nNOS and the PDZ-binding motif of PMCA4b were essential for the protein-protein interaction. The involvement of lipid raft microdomains on the activity of PMCA4b and nNOS was also investigated. Unlike other PMCA isoforms, PMCA4 was relatively more concentrated in the raft fractions. Disruption of lipid rafts altered the intracellular localization of PMCA4b and affected the interaction between PMCA4b and nNOS, which suggest that the unique lipid raft distribution of PMCA4 may be responsible for its regulation of nNOS activity. In summary, lipid rafts may act as platforms for the PMCA4b regulation of nNOS activity and the transient tethering of nNOS to PMCA4b is responsible for rapid nNOS deactivation.

Human urate transporter 1 promoter polymorphisms and hyperuricemia in Chinese Han people / 中华内分泌代谢杂志

ObjectiveTo analyze the association of human urate transporter 1 ( hURAT1 ) gene promoter single-nucleotide polymorphisms(SNPs) with primary hyperuricemia ( HUA ) in Chinese Han people.MethodsA total of 215 patients with HUA and 323 healthy subjects were chosen to be investigated of SNP of hURAT1 promoter by PCR and sequencing.ResultsFive SNPs were identified,including-454A/T,-434T/C,-382C/T,-87C/T,and + 118G/A.Pairwise linkage disequilibrium analysis displayed a high linkage disequilibrium between the five SNPs ( r2 =0.99).In HUA group,the heterozygous genotypos ( AT,CT,CT,CT,AG ) frequencies were significantly lower than those in control group ( P＜0.05 ).Logistic regression analysis showed that the heterozygosis genotypes ( AT,CT,CT,CT,AG) were protective factors of HUA ( OR 0.68-0.75 ).The minor allele ( T,C,T,T,A ) frequencies for both SNPs were significantly different between two groups ( P =0.022,P =0.038 ).ConclusionThese findings indicate that -454A/T,-434T/C,-382C/T,-87C/T,and + 118G/A SNPs of hURAT1 gene promoter area are associated with HUA in Chinese Han population.

Expressions and clinical significance of putative hepatic stem cell biomarkers associated with tumour angiogenesis / 中华普通外科杂志

Objective To investigate the expression of liver cancer stem cells(LCSCs) biomarkers and tumour angiogenesis in patients with hepatocellular carcinoma (HCC) and its clinical significance.Methods Immunohistochemistry was used to detect the expression of CD90,CD133 and CD34 in tumor tissues from 61 patients with HCC undergoing curative hepatectomy.Results The positive rates of CD90and CD133 in HCC tissues were 29.5％ (18/61)and 31.1％ (19/61),the positive rates of CD90 and/or CD133 in HCC were 45.9％ (28/61).The positive rate of LCSCs biomarkers in patients with vascular invasion,tumor differentiation Ⅲ/Ⅳ,non-complete encapsulation,and TNM stage Ⅲ/Ⅳ was significantly higher than those with non-vascular invasion,tumor differentiation Ⅰ / Ⅱ,with complete encapsulation,and TNM stage Ⅰ/Ⅱ.There were a significant positive correlation between LCSCs biomarkers expression and MVD(r =0.5,P ＜ 0.05).As compared with negative LCSCs expression and MVD low group,patients with a positive LCSCs biomarkers expression and MVD high in tumours had significantly lower overall survival (OS) and poor relapse-free survival (RFS).Conclusions High expression levels of LCSCs biomarkers are related to markers of angiogenesis microvessel density (MVD).By combining two of those we can classify paitients and anticipate the prognosis after surgery.