Objective: To design a pair of PCR primers that could identify Dendrobium chrysotoxum specifically, to optimize the system of PCR detection, and to establish a method to identify D. chrysotoxum rapidly and accurately. Methods: From GenBank database, rDNA 170 ITS sequences in the plants of Dendrobium Sw. were downloaded and compared with all sequences using MEGA 5.0; The variation sites were located, and a pair of specific primers in the non-conservative district were designed. PCR amplification was performed using the specific primers with 35 DNA templates in the plants of Dendrobium Sw., D. chrysotoxum was positive. Results: D. chrysotoxum could be specifically amplificated by specific primers when the annealing temperature was raised to 58 °C, while other plants of Dendrobium Sw. were shown as negative, and the sensitivity of the primer could reach 0.69 ng/μL. Conclusion: This study designs a method that could identify D. chrysotoxum specifically. Using this specific primer could identify D. chrysotoxum rapidly and accurately from the homologous species. This method is well-performed in specificity, and it is more simple, convenient, efficient, and accurate than other methods, such as morphological and microscopical identification, chromatograph, and spectral method.

This study provides the candidate sequences in the identification of Radix et Rhizoma Clematidis and its adulterants using DNA barcoding. We amplified and sequenced the region psbA-trnH, with the data of 284 sequences from GenBank, the differential intra- and inter-specific divergences, genetic distance, barcoding gap were used to evaluate five barcodes, and the identification efficiency was assessed using BLAST1 and Nearest Distance methods. The results showed that psbA-trnH barcodes performed high identification efficiency and inter-specific divergences among the five different DNA barcodes. Analysis of the barcoding gap and NJ tree showed psbA-trnH was superior to other barcodes. Based on the identification and PCR amplification efficiency, psbA-trnH can be the ideal barcode to identify Radix et Rhizoma Clematidis and its adulterants accurately.
DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; Drug Contamination ; Nucleic Acid Amplification Techniques ; methods ; Plant Roots ; genetics ; Plants, Medicinal ; classification ; genetics ; Ranunculaceae ; classification ; genetics ; Rhizome ; genetics ; Species Specificity

To establish a new method for identifying genus of Lilium by DNA barcoding technology, ITS, ITS2, psbA-trnH, matK and rbcL sequences were analyzed in term of variation of inter- and intra-species, barcoding gap, neighbor-joining tree to distinguish genus of Lilium based on 978 sequences from experimental and GenBank database, and identification efficiency was evaluated by Nearest distance and BLAST1 methods. The results showed that DNA barcoding could identify different species in genus of Lilium. ITS sequence performed higher identification efficiency, and had significant difference between intra- and inter-species. And NJ tree could also divide species into different clades. Results indicate that DNA barcoding can identify genus of Lilium accurately. ITS sequence can be the optimal barcode to identify species of Lilium.
DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Lilium ; classification

Objective: To analyze the effect of acupuncture versus hormone replacement therapy (HRT) for premature ovarian insufficiency (POI). Methods: China National Knowledge Infrastructure (CNKI), Wanfang Academic Journal Full-text Database (Wanfang), Chongqing VIP Database (CQVIP), China Biology Medicine Disc (CBM), Web of Science, Cochrane Library, PubMed, and Excerpta Medica Database (EMBASE) were searched up to January 31st, 2019 to identify randomized controlled trials (RCTs) evaluating the effect of acupuncture for POI. The primary outcome was the level of basal serum follicle- stimulating hormone (FSH). Secondary outcomes included serum levels of luteinizing hormone (LH), estradiol (E2) and anti-Müllerian hormone (AMH). Two authors extracted data independently and assessed the risk of bias and the methodological quality using the Cochrane's tool. Meta-analysis was conducted by RevMan version 5.3. Results: Eight eligible RCTs with a total of 496 POI patients were included in the meta-analysis. The pooled results showed that there was a significant reduction in the basal serum FSH level (MD=-5.82, 95％CI:-9.76 to -1.87, I2=82％, P=0.004) and a remarkable elevation in the basal E2 level (SMD=0.93, 95％CI: 0.34 to 1.52, I2=88％, P=0.002) in the acupuncture group when compared with the control. Subgroup analysis showed that compared with HRT, a significant decrease in the FSH level was observed in both acupuncture alone (MD=-4.53, 95％CI:-8.96 to -0.10, I2=73％, P=0.04) and acupuncture plus HRT (MD=-9.60, 95％CI:-17.60 to -1.61, I2=50％, P=0.02), while a remarkable elevation of E2 was only found in acupuncture plus HRT (SMD=1.43, 95％CI: 1.03 to 1.82, I2=0％, P<0.00001). There was no significant difference in the LH level between acupuncture and HRT (MD=-3.16, 95％CI:-9.41 to 3.10, I2=0％, P=0.32), only one trial reported AMH, and no significant difference was found between acupuncture and HRT. Conclusion: The present study indicated that acupuncture had an advantage over HRT in reducing serum FSH level and increasing serum E2 level in women with POI. However, evidence supporting the finding is limited due to the small sample size, potential methodological flaws and significant heterogeneity. Hence, this conclusion still needs to be verified by high-quality RCTs.

OBJECTIVET To explore the relationship between the expression of SOX4 gene and early recurrence of hepatocellular carcinoma (HCC) after curative resection.

METHODSSOX4 expression was detected immunohistochemically in 60 HCC patients including 30 with and 30 without early recurrence after curative resection, with 30 normal liver specimens as the control.

RESULTSThe expression of SOX4 was significantly higher in HCC than in normal liver (41.7% vs 16.7%, P<0.05), and in HCC tissues, the expression was significantly higher in early recurrent HCC after curative resection than in HCC without early recurrence (56.7% vs 26.7%, P<0.05). SOX4 expression was inversely correlated to the patients' gender, age, tumor size, HBsAg, and Edmonson grade (P<0.05).

CONCLUSIONSOX4 is closely associated with early recurrence of HCC after curative resection, and its overexpression may contribute to early recurrence of HCC. SOX4 may serve as a new molecular indicator for evaluating the prognosis of HCC.

Adult ; Aged ; Biomarkers, Tumor ; Carcinoma, Hepatocellular ; genetics ; metabolism ; surgery ; Female ; Humans ; Liver Neoplasms ; genetics ; metabolism ; surgery ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; genetics ; Prognosis ; SOXC Transcription Factors ; genetics ; metabolism

A large amount of nicotinamide adenine dinucleotide phosphate (NADPH) is required for fatty acid synthesis and maintenance of the redox state in cancer cells. Malic enzyme 1(ME1)-dependent NADPH production is one of the three pathways that contribute to the formation of the cytosolic NADPH pool. ME1 is generally considered to be overexpressed in cancer cells to meet the high demand for increased de novo fatty acid synthesis. In the present study, we found that glucose induced higher ME1 activity and that repressing ME1 had a profound impact on glucose metabolism of nasopharyngeal carcinoma(NPC) cells. High incorporation of glucose and an enhancement of the pentose phosphate pathway were observed in ME1-repressed cells. However, there were no obvious changes in the other two pathways for glucose metabolism: glycolysis and oxidative phosphorylation. Interestingly, NADPH was decreased under low-glucose condition in ME1-repressed cells relative to wild-type cells, whereas no significant difference was observed under high-glucose condition. ME1-repressed cells had significantly decreased tolerance to low-glucose condition. Moreover, NADPH produced by ME1 was not only important for fatty acid synthesis but also essential for maintenance of the intracellular redox state and the protection of cells from oxidative stress. Furthermore, diminished migration and invasion were observed in ME1-repressed cells due to a reduced level of Snail protein. Collectively, these results suggest an essential role for ME1 in the production of cytosolic NADPH and maintenance of migratory and invasive abilities of NPC cells.
Carcinoma ; Cell Line, Tumor ; Cell Movement ; Cell Survival ; Glucose ; metabolism ; Glycolysis ; Humans ; Malate Dehydrogenase ; metabolism ; NADP ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness ; Oxidation-Reduction ; Oxidative Phosphorylation ; Pentose Phosphate Pathway ; Proto-Oncogene Proteins c-akt ; metabolism

AIMTo determine whether repetitive exposure to high sustained +Gz acceleration induces persisting changes in the myocardial free radical metabolism and observe the protective effects of low-G training and antioxidant tea polyphenols (TP).

METHODSThirty-two male Wistar rats were randomly divided into four groups (n=8 each): group A, restrained, was only submitted to +1 Gz for 5 min. Group B, centrifuged, was exposed to five plateaus of 30 s at +10 Gz for intermittent times, three times a week, for three weeks. Group C, low-G trained, was exposed to +2 Gz for 5 min about 1 h prior to +10 Gz stress, and group D was orally given TP at dose of 200 mg/kg about 1 h prior to +10 Gz stress. On the next day morning after last centrifuge run, the rats were decapitated and the hearts were quickly removed. Malondialdehyde (MDA) concentration and superoxide dismutase (SOD) activity were measured. Additionally, CuZn-SOD and inducible NO synthase (iNOS) enzymatic contents were examined by immunohistochemical staining and their mRNA were analyzed by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR).

RESULTSCompared with group A, MDA concentration and iNOS enzymatic content in myocardial mitochondria were increased significantly (P < 0.05) in group B. Compared with group B, mitochondrial SOD activity was significantly increased in group C (P < 0.05). iNOS enzymatic content was significantly decreased in group C and D. There were no significant differences of CuZn-SOD content, CuZn-SOD and iNOS mRNA levels among the four groups.

CONCLUSIONRepeated high +Gz exposure can induce myocardial free radical metabolic disorder and mainly result in mitochondrial peroxidative injury. But low-G training and natural antioxidant TP have protective effects, and the former is better.

Acceleration ; Adaptation, Physiological ; physiology ; Animals ; Free Radicals ; metabolism ; Male ; Myocardium ; metabolism ; Polyphenols ; pharmacology ; Rats ; Rats, Wistar ; Tea ; chemistry

Diabetic retinopathy(DR)is a prevalent microvascular complication of diabetes and the leading cause of blindness and severe visual impairment in adults.The high levels of glucose trigger multiple intracellular oxidative stress pathways,such as POLDIP2,resulting in excessive reactive oxygen species(ROS)pro-duction and increased expression of vascular cell adhesion molecule-1(VCAM-1),hypoxia-inducible factor 1α(HIF-1α),and vascular endothelial growth factor(VEGF),causing microvascular dysfunction.Dihydromyricetin(DMY)is a natural flavonoid small molecule antioxidant.However,it exhibits poor solubility in physiological environments,has a short half-life in vivo,and has low oral bioavailability.In this study,we present,for the first time,the synthesis of ultra-small Fe-DMY nano-coordinated polymer particles(Fe-DMY NCPs),formed by combining DMY with low-toxicity iron ions.In vitro and in vivo experiments confirm that Fe-DMY NCPs alleviate oxidative stress-induced damage to vascular endo-thelial cells by high glucose,scavenge excess ROS,and improve pathological features of DR,such as retinal vascular leakage and neovascularization.Mechanistic validation indicates that Fe-DMY NCPs can inhibit the activation of the Poldip2-Nox4-H2O2 signaling pathway and downregulate vital vascular function indicators such as VCAM-1,HIF-1α,and VEGF.These findings suggest that Fe-DMY NCPs could serve as a safe and effective antioxidant and microangio-protective agent,with the potential as a novel multimeric drug for DR therapy.

OBJECTIVETo develop quality of life questionnaire of Chinese medicine for postoperative patients with colorectal cancer (QLQ-CMPPCC), thus comprehensively and objectively evaluating the clinical efficacy of Chinese medicine and pharmacy in treating postoperative patients with colorectal cancer (CC).

METHODSThe theoretical structure model of the questionnaire was addressed in combined with basic theories of Chinese medicine according to the principle of WHO quality of life (QOL). The primary questionnaire was developed using methods of structuralization policy making after we extensively retrieve various universal and specific questionnaires for CC cancer patients at home and abroad. The 205 CC patients were tested by questionnaire. The items were screened using experts grading method, item selection analysis, dispersion trends of standard deviation, t-test, correlation coefficient method, factor analysis,and Cronbach's alpha.

RESULTSThe QLQ-CMPPCC was developed containing four domains of physical, psychological, independence, and social functions, involving 20 aspects and 54 items. Of them, non-fistula patients answered 43 items and fistula patients answered 46 items. One item covered the general QOL evaluation.

CONCLUSIONSQLQ-CMPPCC showed Chinese medical features. It comprehensively reflected the connotation of QOL for postoperative CC patients. It could be taken as a tool for evaluating Chinese medical efficacy for postoperative CC patients.

Colorectal Neoplasms ; surgery ; Humans ; Medicine, Chinese Traditional ; methods ; Postoperative Period ; Quality of Life ; Surveys and Questionnaires ; Treatment Outcome

Functional magnetic resonance imaging (fMRI) was used to study the activated brain areas of human during simple and complex digital calculation, and to investigate the role of cortical and subcortical structures involved in the mental calculation. Sixteen right-handed healthy volunteers performed mental calculation of simple and complex addition/subtraction respectively, while the fMRI data were recorded by a Seimens 1.5 T MR machine. Block-design was used in the tasks. Two calculation tasks and one base-line tasks were performed for the block-design. Simple calculation task was single-digit addition and subtraction, while the complex was multi-digit addition and subtraction. The base-line task was to tell whether the two numbers were the same in every trial. Statistical parametric mapping (SPM99) was employed to process data and localize functional areas. We compared the average activation intensity of each activated brain regions in the same calculation task and the activation intensity of the same regions in both tasks respectively. Both the cortex and the subcortical structures including basal ganglia and thalamus were activated during simple and complex mental calculations. Similar brain regions in subjects including frontal lobe, parietal lobe, occipital lobe, cingulate gyrus, thalamus and cerebellum were engaged in simple and complex addition/subtraction. In the same task, activation intensity of all activated brain areas differed insignificantly. Compared with the complex task, the right parietal lobe was not activated in the simple one. The subcortical structures such as the caudate nucleus and the left marginal division of the striatum (MrD) were activated in both two calculation tasks. The cortical regions involved in both simple and complex addition/subtraction were similar. In conclusion, both the cortex and the subcortical structures were activated during the mental calculation. The cortex including the frontal cortex, parietal cortex, and cingulate gyrus were activated during mental calculation, while the subcortical structures such as the caudate nucleus, the globus pallidum and the left marginal division of the striatum also played a critical role in the neural networks of the calculation at the same time. Right parietal lobe (supramarginal gyrus) was engaged only in the complex task, which suggested that this region might be involved in the visuospatial memory and processing.
Brain ; physiology ; Brain Mapping ; Humans ; Magnetic Resonance Imaging ; Parietal Lobe ; physiology ; Thinking ; physiology