Prognosis of Diabetic Peripheral Neuropathy via Decomposed Digital Volume Pulse from the Fingertip.

Diabetic peripheral neuropathy (DPN) is a very common neurological disorder in diabetic patients. This study presents a new percussion-based index for predicting DPN by decomposing digital volume pulse (DVP) signals from the fingertip. In this study, 130 subjects (50 individuals 44 to 89 years of age without diabetes and 80 patients 37 to 86 years of age with type 2 diabetes) were enrolled. After baseline measurement and blood tests, 25 diabetic patients developed DPN within the following five years. After removing high-frequency noise in the original DVP signals, the decomposed DVP signals were used for percussion entropy index (PEIDVP) computation. Effects of risk factors on the incidence of DPN in diabetic patients within five years of follow-up were tested using binary logistic regression analysis, controlling for age, waist circumference, low-density lipoprotein cholesterol, and the new index. Multivariate analysis showed that patients who did not develop DPN in the five-year period had higher PEIDVP values than those with DPN, as determined by logistic regression model (PEIDVP: odds ratio 0.913, 95% CI 0.850 to 0.980). This study shows that PEIDVP can be a major protective factor in relation to the studied binary outcome (i.e., DPN or not in diabetic patients five years after baseline measurement).

Activation of liver X receptor suppresses osteopontin expression and ameliorates nephrolithiasis.

Nephrolithiasis is a common disease of the urinary system, of which idiopathic calcium oxalate (CaOx) kidney stones, in particular, are one of the special types. In the initial stages of CaOx kidney stone formation, Randall's plaques (RPs) develop. Liver X receptors (LXRs) inhibit oxidative stress and inflammatory in other diseases; nevertheless, the role of LXRs in nephrolithiasis has yet to be elucidated. In this study, the role of LXRs in the progression of RP formation was investigated. Microarray analysis revealed that LXR/RXR levels were significantly greater in low-plaque tissues (<5%) than in high-plaque tissues (>5%), confirming the link between LXR activation and RP formation. Correspondingly, expression levels of two LXR target genes, LXRα and LXRß, were lower in high-plaque tissues than in low-plaque tissues. In vitro, LXR agonist alleviated calcium oxalate monohydrate-induced cellular calcium deposits and apoptosis. LXR activation decreased reactive oxygen species production and gene expression of inflammatory mediators, including osteopontin that has recently been demonstrated to correlate with the development of RPs. Moreover, p38 MAPK and JNK signaling may mediate LXR-regulated expression in HK-2 cells. In an animal model, the deposition was reduced by activating LXR, and osteopontin expression was also inhibited. Our findings suggest a role for LXRs in the progression of idiopathic CaOx kidney stones; LXR agonists may have therapeutic potential for the treatment of nephrolithiasis.

Comparison of Three Methods Estimating Baseline Creatinine For Acute Kidney Injury in Hospitalized Patients: a Multicentre Survey in Third-Level Urban Hospitals of China.

BACKGROUND/AIMS: A lack of baseline serum creatinine (SCr) data leads to underestimation of the burden caused by acute kidney injury (AKI) in developing countries. The goal of this study was to investigate the effects of various baseline SCr analysis methods on the current diagnosis of AKI in hospitalized patients. METHODS: Patients with at least one SCr value during their hospital stay between January 1, 2011 and December 31, 2012 were retrospectively included in the study. The baseline SCr was determined either by the minimum SCr (SCrMIN) or the estimated SCr using the MDRD formula (SCrGFR-75). We also used the dynamic baseline SCr (SCrdynamic) in accordance with the 7 day/48 hour time window. AKI was defined based on the KDIGO SCr criteria. RESULTS: Of 562,733 hospitalized patients, 350,458 (62.3%) had at least one SCr determination, and 146,185 (26.0%) had repeat SCr tests. AKI was diagnosed in 13,883 (2.5%) patients using the SCrMIN, 21,281 (3.8%) using the SCrGFR-75 and 9,288 (1.7%) using the SCrdynamic. Compared with the non-AKI patients, AKI patients had a higher in-hospital mortality rate regardless of the baseline SCr analysis method. CONCLUSIONS: Because of the scarcity of SCr data, imputation of the baseline SCr is necessary to remedy the missing data. The detection rate of AKI varies depending on the different imputation methods. SCrGFR-75 can identify more AKI cases than the other two methods.

[Research on ginseng trade competitiveness between China and Korea].

Ginseng is one of China's valuable Chinese herbal medicines, with a long using history. Ginseng has worldwide reputation, and widely used in food, medicine, health products, cosmetics and other production. China and South Korea have a big ginseng industrial, and sharing half of the export market. The ginseng export competitiveness analysis seems important and necessary between China and South Korea. In this paper, the data of customs and trade of ginseng in COMTRADE database were studied, and ginseng export competitiveness was analyzed between China and Korea. The results showed that the ginseng export competitiveness of Korean more competitive than China. Contrast with China, South Korea using only 15% total amount of ginseng exports and produced the same total export amount. This article has the reference value to the traditional Chinese medicine resources management and the economics research. On this basis, this paper further discusses the problems that should be paid attention to in the development of ginseng industry in China.

A Tumor-Specific Neo-Antigen Caused by a Frameshift Mutation in BAP1 Is a Potential Personalized Biomarker in Malignant Peritoneal Mesothelioma.

Malignant peritoneal mesothelioma (MPM) is an aggressive rare malignancy associated with asbestos exposure. A better understanding of the molecular pathogenesis of MPM will help develop a targeted therapy strategy. Oncogene targeted depth sequencing was performed on a tumor sample and paired peripheral blood DNA from a patient with malignant mesothelioma of the peritoneum. Four somatic base-substitutions in NOTCH2, NSD1, PDE4DIP, and ATP10B and 1 insert frameshift mutation in BAP1 were validated by the Sanger method at the transcriptional level. A 13-amino acids neo-peptide of the truncated Bap1 protein, which was produced as a result of this novel frameshift mutation, was predicted to be presented by this patient's HLA-B protein. The polyclonal antibody of the synthesized 13-mer neo-peptide was produced in rabbits. Western blotting results showed a good antibody-neoantigen specificity, and Immunohistochemistry (IHC) staining with the antibody of the neo-peptide clearly differentiated neoplastic cells from normal cells. A search of the Catalogue of Somatic Mutations in Cancer (COSMIC) database also revealed that 53.2% of mutations in BAP1 were frameshift indels with neo-peptide formation. An identified tumor-specific neo-antigen could be the potential molecular biomarker for personalized diagnosis to precisely subtype rare malignancies such as MPM.

[Water extract from Codonopsis thalictrifolia wall affects the reproductive system of male infant rats].

OBJECTIVE: To study the impact of the water extract from Codonopsis thalictrifolia Wall (CTW) on the reproductive METHODS: We divided 32 male SD infant rats into four groups of equal number to be treated intragastrical-system of male infant rats. ly with distilled water (control) and CTW at 10 g/kg (low dose) , 20 g/kg (medium dose), and 40 g/kg (high dose), respectively, twice a day for 2 weeks. Then we killed the rats, measured the levels of testosterone (T), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in the serum, obtained the testis weight, body weight, testis visceral coefficient and sperm concentration, and detected sperm viability, sperm motility and the level of cyclic adenosine monophosphate (cAMP) in the Leydig cells, followed by RESULTS: Compared with the control group, the low-dose, me-analysis of differences among different groups using the SPSS software. Medium-dose and high-dose CTW groups showed significant decreases in the serum T level ([3.09 +/-0.42] vs [1.22 +/-0. 32] , [1.06 +/- 0.29] and [0.57 +/-0.18] nmol/L, P<0.01), testis weight ([1.40 +/-0.16] vs [0.96 +/-0.09], [0.92 +/-0.11] and [0.91 +/- 0.08] g, P <0.01), and sperm concentration ([1.03 +/-0.16] vs [0.19 +/-0.07], [0.17 +/-0.08] and [0.16 +/-0.07] x 10(6)/ml, P <0.01), but a dramatic elevation in the testis visceral coefficient ([42.22 +/- 3.02] vs [51.39 +/- 3.09], [52.28 +/- 4.86] and [54.13 +/-6.06] mg/10 g, P <0.01); the medium- and high-dose CTW groups exhibited remarkable increases in the levels of serum LH ([13.62+/-0.89] vs [14.69 +/-0.12] and [14.93 +/-0.28] ng/L, P<0.01) and FSH ([4.32 +/-0.18] vs [4.77 +/-0.23] and [4.89 +/-0. 38] IU/L, P <0.05); all the three CTW groups showed markedly inhibited serum T secretion ([1.85 +/- 0.18] vs [1.42 +/-0.15], [1.12+/-0.18] and [0.88 +/-0.21] nmol/L, P<0.01) and intracellular cAMP ([5.51 +/-0.12] vs [4.39+/-0.06], [4.28 +/-0.07] and [4.11 +/- 0.10] nmol/L, P <0.01) in the Leydig cells. CONCLUSION: The water extract from CTW may reduce the synthesis of testosterone in the serum of male infant rats through the PKA pathway and consequently inhibit their testicular development and sperm production and affect the development of their reproductive system.

Engineering and refolding of a novel trimeric fusion protein TRAIL-collagen XVIII NC1.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered to be a promising anticancer agent because its active form TRAIL trimer is able to induce apoptosis in different tumor cell lines while sparing normal cells. However, TRAIL trimer possesses a short half-life and low stability, which turns out to be a major obstacle for the development of clinical trials. In our present study, we constructed a recombined TRAIL trimer by genetic fusion of non-collagenous domain (NC1) of human collagen XVIII or its trimerization domain (TD) to C-terminus of TRAIL via a flexible linker, and then refolded the fusion proteins using a two-step refolding approach, namely a combination of dilution and gel filtration chromatography. As a result, both recombinant proteins, TRAIL-NC1 and TRAIL-TD, were expressed in Escherichia coli as inclusion bodies, and they exhibited difficultly to refold efficiently by conventional methods. Thereby, we applied a modified two-step refolding approach to refold fusion proteins. More than 55 % of TRAIL-NC1 and 90 % of TRAIL-TD protein activity was recovered during the two-step refolding approach, and their stability was also increased significantly. Also, size exclusion chromatography showed refolded TRAIL-NC1 was a trimer while TRAIL-TD, hexamer. However, both of them exerted good apoptosis activity on NCI-H460 cells.

Effect of Sheng Xue Ning Tablets on Renal Anemia in Patients Subject to Maintenance Hemodialysis and Safety Evaluation: A Multi-setting Prospective Randomized Study.

This study compared Sheng Xue Ning (SXN) tablets with ferrous succinate (FS) tablets in terms of their efficacy for the treatment of iron-deficient renal anemia and safety in patients subject to maintenance hemodialysis (MHD). A total of 94 patients undergoing MHD were randomly assigned to an experiment group (receiving oral SXN tablets, SXN group) and a control group (orally given FS tablets, FS group) and followed up for 12 weeks. Erythropoietin (EPO) was used in both groups. The efficacy was assessed by detecting the subsequent changes in hemoglobin (Hb), serum iron (SI), SF and transferrin saturation (TSAT). At the 12th week, Hb and TSAT levels in both groups were significantly increased compared to those in the screening period (P<0.05). However, no significant difference in Hb and TSAT was found between the two groups. The average weekly EPO dosage used was lower in SXN group than in FS group (P<0.05) at the 10th week and the 12th week. Our study showed that SXN tablets can effectively ameliorate renal anemia and keep iron metabolism stable in MHD patients, and its efficacy is virtually close to that of FS tablets. Meanwhile, SXN tablets can reduce the dosage of EPO and have a good safety profile.

Percussion Entropy Analysis of Synchronized ECG and PPG Signals as a Prognostic Indicator for Future Peripheral Neuropathy in Type 2 Diabetic Subjects.

Diabetic peripheral neuropathy (DPN) is one of the most common chronic complications of diabetes. It has become an essential public health crisis, especially for care in the home. Synchronized electrocardiogram (ECG) and photoplethysmography (PPG) signals were obtained from healthy non-diabetic (n = 37) and diabetic (n = 85) subjects without peripheral neuropathy, recruited from the diabetic outpatient clinic. The conventional parameters, including low-/high-frequency power ratio (LHR), small-scale multiscale entropy index (MEISS), large-scale multiscale entropy index (MEILS), electrocardiogram-based pulse wave velocity (PWVmean), and percussion entropy index (PEI), were computed as baseline and were then followed for six years after the initial PEI measurement. Three new diabetic subgroups with different PEI values were identified for the goodness-of-fit test and Cox proportional Hazards model for relative risks analysis. Finally, Cox regression analysis showed that the PEI value was significantly and independently associated with the risk of developing DPN after adjustment for some traditional risk factors for diabetes (relative risks = 4.77, 95% confidence interval = 1.87 to 6.31, p = 0.015). These findings suggest that the PEI is an important risk parameter for new-onset DPN as a result of a chronic complication of diabetes and, thus, a smaller PEI value can provide valid information that may help identify type 2 diabetic patients at a greater risk of future DPN.

[Authentication of Pinellia pedatisecta and their adulterants by AS-PCR].

OBJECTIVE: To establish a molecular method for the authentication of Pinellia pedatisecta and its adulterants. METHOD: DNA sequences of some species from P. tenore, Typhonium and Arisaema were downloaded from GenBank, the sequences were aligned using DNAMAN. Allele-specific primers for P. pedatisecta and P. tenore were designed according to their SNPs in rpl 20 sequence. The designed primers were used to amplify 10 samples of P. pedatisecta, P. ternata and T. flagelliforme. RESULT: A 351 bp band was amplified from P. pedatisecta but not form P. ternata and T. flagelliforme by primer Pprpl149F and Pprpl484R. A 630 bp band was amplified from P. ternate and P. pedatisecta but not from T. flagelliforme by primer Ptrpl94F and Ptrpl699R. CONCLUSION: AS-PCR has the advantages of highly specific and good reproducibility, by which P. pedatisecta can be identified from part of its adulterants quickly. It is a potential method to be used in the molecular identification of other materia medica.

[A new molecular identification method: anchored primer amplification polymorphism DNA].

To build up a stable and easy doing method for molecular identification in traditional Chinese medicine, on basis of RAPD, the new method mainly changed the primer length and PCR annealing temperature. Panax ginseng, Panax quinquefolius and its nine adulterants were used to establish the method and test it using MARMS primers published in 2004. The new method also used to authenticate Chinese Materia Medica of Tian-hua-fen (Radix Trichosanthes) and Bai-zhi (Radix Angelica). Primer Pg-q36F obtained polymorphic bands of P. Ginseng, P. quinquefolius and its adulterants. The identification result is identical to that published before and more stable. Primer TkS1-64F obtained polymorphic bands of Tian-hua-fen and its nine adulterants. Primer AfS1-100F obtained polymorphic bands of Bai-zhi and its three adulterants. The method has good stability and reproducibility and can easily identify authertic medicines from their adulterants. It was a potential molecular method to identify other Chinese Materia Medica. The method was named as anchored primer amplification polymorphism DNA (APAPD).

[Research on characterization analysis of synchronous fluorescence spectra of living phytoplankton].

The synchronous fluorescence spectra of five phytoplankton species growing under three temperatures (25, 20 and 15 degrees C)and three illuminations (7000, 4100 and 1100 Lx)were measured and processed by multinomial smoothness and autoscaling to obtain the characteristic spectra. Principal component analysis was used to obtain standard spectra. The analysis shows that at different temperatures, the characteristic spectra of Skeletonema costatuma, Isochrysis galbana, and Platymonas helgolanidica show high similarities, while the spectra similarities of Alexandrium tamarense and Gymnodinium stein are not as good as the above three species. The standard spectrum of Skeletonema costatuma, which belongs to Bacillariophyta, is quite different from those of Alexandrium tamarense and Gymnodinium stein, which belong to Dinophyta.

[A new method to obtain molecular marker of sequence-tagged site (STS) of Panax ginseng and P. quinquefolius].

OBJECTIVE: Searching a new molecular method to authenticate Panax ginseng and P. quenquefolium. METHOD: Single primers based on rDNA sequences of Panax species were designed to obtain polymorphic bands of P. ginseng and P. quinquefolius and then sequenced. Four PCR primers (two forword and two reverse primers) specific to P. ginseng and P. quinquefolius were designed. RESULT: Primer Pg-6F, Pg-479R only amplified 474 bp band for P. ginseng and primer Pq-442F, Pq-658R only amplified 217 bp band for P. quinquefolius. It is indicated that the four primers could serve as specific STS primers for Panax species. CONCLUSION: A new way to obtain STS primers of Panax species was established. This method is more quick and efficient than SCAR-PCR method and can serve as a model to obtain molecular markers for other Chinese material medica.

[Functional genomics studies of Salvia miltiorrhiza I. Establish cDNA microarray of S. miltiorrhiza].

OBJECTIVE: Establishing cDNA microarray, in order to study functional genomics of Salvia miltiorrhiza. METHOD: Total RNA samples were prepared from S. miltiorrhiza roots using a modified CTAB method. mRNA was isolated by Quichprep Micro mRNA Purification Kit from Pharmacia. Then cDNA was synthesized and cloned into the EcoRI-XhoI sites of the ZAP Express vector using a cDNA synthesis kit, and the ligation mixture was packaged using a ZAP-cDNA Gigapack Gold III cloning kit (Stratagene). The single phage was isolated for PCR amplification, Aliquots of the PCR reactions were analyzed in a 1.5% agarose gel to verify the quality of PCR. The remaining cDNA was purified by Multiscreen filter plates (Millipore) and aliquots were analyzed by agarose gel again to verify the quality of purification. Clones passed verification was resuspended in 15 microL 50% DMSO for arraying. An actin gene from S. miltiorrhiza was used for positive control. PloyA and 50% DMSO was used for negative controls. RESULT: Bacterial colonies containing cNDAs of S. miltiorrhiza were inserted with average insert size of 0. 5 kb to 2. 5 kb. Total 4 354 genes were singled out from the first 8 736 PCR product and used for cDNA microarray manufacture. Single color fluorescence hybridization showed that all positive controls had signals while negative controls had no signals. CONCLUSION: It was the first cDNA microarray about traditional Chinese herbs especially for geoherbs. It could be a powerful tool for studying functional genomics of S. miltiorrhiza.

[Application of multiplex allele-specific PCR for authentication of Panax ginseng and P. quinquefolius].

OBJECTIVE: Searching and identifying SNP in Panax species and using multiplex allele-specific PCR (MAS-PCR) to authenticate P. ginseng and P. quenquefolium. METHOD: Based on genbank database of Panax species, using DNAMAN to align the sequences, identify SNP of P. ginseng and P. quenquefolium. Design allele-specific primers for P. ginseng and P. quenquefolium, optimize the PCR reaction system including the usage amount of Taq, dNTP, primer, etc. Optimized system was performed with the total DNA of 20 different sources of P. ginseng and P. quenquefolium. RESULT: When the annealing temperature was 66 'C, the template DNA of P. ginseng could be amplified 249 bp band whereas P. quenquefolium amplified 1 049 bp band. CONCLUSION: The MAS-PCR have the advantages of highly specific, good reproducibility and could be identify P. ginseng and P. quenquefolium in the same PCR tube. It was a potential method to use in the molecular identification of other meteria medica.

[High specific PCR identification of Bungarus multicinctus and its adulterants].

OBJECTIVE: To develop a convenient and effective method for the identification of Bungarus multicinctus. METHOD: Based on the sequence of Cyt b gene fragment of B. multicinctus and its adulterants, a pair of highly specific primer (HJL- and HJH-) were designed for distinguishing B. ulticinctus from other species of snake. To establish specific PCR reaction condition, the primers were employed to amplify the DNA templates extracted from B. multicinctus and 6 other species of snake, under different annealing temperature. Using this method, B. multicinctus was identified from 18 samples bought from many drugstores. RESULT: A 230 bp DNA fragment was amplified from B. multicinctus in PCR with annealed temperature at 67 degrees C, whereas no DNA fragment was amplified from other snake samples under the same reaction condition, B. multicinctus could be clearly distinguished from others by PCR reaction with the highly specific primers. In the present study, 18 sample, bought from different drugstores, were also identified by the highly specific PCR with the primers. The results indicated that 14 samples were B. multicinctus and the other 4 were adulterant, which was consistent with the conclusion of authentication based on morphological. CONCLUSION: The primers designed in the present study were highly specific for B. multicinctus.

Novel conjugation of tumor-necrosis-factor-related apoptosis-inducing ligand (TRAIL) with monomethyl auristatin E for efficient antitumor drug delivery.

Monomethyl auristatin E (MMAE) is conjugated with TNF-related apoptosis-inducing ligand (TRAIL) via a linker that is stable in extracellular fluid, while it is cleaved by cathepsin once the conjugate has entered a tumor cell, thus activating the antimitotic mechanism of MMAE. The TRAIL-MMAE conjugate is a conceptually viable therapeutic strategy with improved in vitro antitumor activity, cell circle arrest and specific accumulation in tumor to treat TRAIL-resistant tumors.