Simultaneous and Visual Detection of KPC and NDM Carbapenemase-Encoding Genes Using Asymmetric PCR and Multiplex Lateral Flow Strip.

Carbapenem-resistant Enterobacteriaceae (CRE) infections constitute a threat to public health, and KPC and NDM are the major carbapenemases of concern. Rapid diagnostic tests are highly desirable in point-of-care (POC) and emergency laboratories with limited resources. Here, we developed a multiplex lateral flow assay based on asymmetric PCR and barcode capture probes for the simultaneous detection of KPC-2 and NDM-1. Biotinylated barcode capture probes corresponding to the KPC-2 and NDM-1 genes were designed and cast onto two different sensing zones of a nitrocellulose membrane after reacting with streptavidin to prepare a multiplex lateral flow strip. Streptavidin-coated gold nanoparticles (SA-AuNPs) were used as signal reporters. In response to the target carbapenemase genes, biotin-labelled ssDNA libraries were produced by asymmetric PCR, which bond to SA-AuNPs via biotin and hybridise with the barcode capture probe via a complementary sequence, thereby bridging SA-AuNPs and the barcode capture probe to form visible red lines on the detection zones. The signal intensities were proportional to the number of resistance genes tested. The strip sensor showed detection limits of 0.03 pM for the KPC-2 and 0.07 pM for NDM-1 genes, respectively, and could accurately distinguish between KPC-2 and NDM-1 genes in CRE strains. For the genotyping of clinical isolates, our strip exhibited excellent consistency with real-time fluorescent quantitative PCR and gene sequencing. Given its simplicity, cost-effectiveness, and rapid analysis accomplished by the naked eye, the multiplex strip is promising auxiliary diagnostic tool for KPC-2 and NDM-1 producers in routine clinical laboratories.

[Effects of aridity on carbon, nitrogen, and phosphorus contents in soils and plants of mountain swamps, Zhejiang, China]. / 旱化对浙江山地沼泽湿地土壤与植物碳氮磷含量的影响.

Mountain swamps in Zhejiang Province have been suffered from severe drouhgt threats because of climate change and artificial interruption. Sphagnum bogs and swamps were gradually degraded into arid swamps. However, the effects of arid processes on the C, N, P contents of soils and their stoichiometry in mountain swamps are still unclear. We measured C, N and P concentrations, pH values, and bulk density in the upper 0-60 cm soil layers in the stands of five mountain swamps with the different arid levels. Moreover, the aboveground biomass and the C, N, P concentrations in the crushed plant mixture were also measured. The results showed that the soils of mountain swamps in Zhejiang Province were rich in soil organic carbon (SOC), total nitrogen (TN), but infertile in phosphorus (TP). Aboveground biomass, soil pH, bulk density increased, while SOC, TN, TP, C:N, C:P, N:P decreased with aridity. Soil pH and bulk density had significant negative correlations with SOC, TN, and TP in soils, respectively. The differences in the C, N, P accumulation in the soils were probably associated with litterfall production, the oxygen condition of wetlands, and the degree of plant decomposition at the different types of mountain swamps. In all, arid trends were obvious at the mountain swamps in Zhejiang Province. Soil nutrients, such as C, N, P, deceased, while plant community succeeded from the wet swamp to the mesophyte vegetation with the arid processes. The contents of C, N and P in the plants varied across species, and were not coupled with those in the soils.

Observation of an electronic order along [110] direction in FeSe.

Multiple ordered states have been observed in unconventional superconductors. Here, we apply scanning tunneling microscopy to probe the intrinsic ordered states in FeSe, the structurally simplest iron-based superconductor. Besides the well-known nematic order along [100] direction, we observe a checkerboard charge order in the iron lattice, which we name a [110] electronic order in FeSe. The [110] electronic order is robust at 77 K, accompanied with the rather weak [100] nematic order. At 4.5 K, The [100] nematic order is enhanced, while the [110] electronic order forms domains with reduced correlation length. In addition, the collective [110] order is gaped around [-40, 40] meV at 4.5 K. The observation of this exotic electronic order may shed new light on the origin of the ordered states in FeSe.

Effect of traditional Chinese medicine comprehensive nursing on senile patients with coronary heart disease and constipation of qi stagnation and blood stasis type / 中国实用护理杂志

Objective:To explore the application effect of traditional Chinese medicine comprehensive nursing on elderly patients with coronary heart disease(qi stagnation and blood stasis type) constipation.Methods:A total of 126 elderly patients with coronary heart disease (qi stagnation and blood stasis type) constipation were divided into the control group and the experimental group by random digits table method, 63 cases each; the control group received routine nursing, and the experimental group was given traditional Chinese medicine comprehensive nursing; comparing the two groups of patients defecation, mental state, recurrence rate, and nursing satisfaction.Results:There was no significant difference in defecation frequency, each defecation time, defecation smoothness score, fecal trait score, fall inflated score, and self-Rating Anxiety Scale (SAS) score( P >0.05). After one month of nursing, defecation frequency, each defecation time, defecation smoothness score, fecal trait score, fall inflated score, and SAS score were (4.09±0.86) times/week, (5.12±1.24) min/time, (2.57±0.39) points, (1.53±0.31) points, (0.97±0.21) points, (49.27±4.15)points in the experimental group, (3.12±0.61) times/week, (6.87±1.52) min/time, (2.59±0.36) points, (2.02±0.44) points, (1.29±0.27) points, (54.42±6.01) points in the control group, the differences were significant ( t values were -6.638-7.081, all P<0.05). Within 1 month after nursing, the recurrence rate of constipation in the experimental group (14.3%, 9/63) was lower than that in the control group (74.6%, 47/63) , and the satisfaction rate of the experimental group (98.4%, 62/63) was higher than that in the control group (69.8%, 44/63) , the differences were significant(× 2 values were 46.414, 19.257, P<0.05). Conclusions:Traditional Chinese medicine comprehensive care for elderly patients with coronary heart disease (qi stagnation and blood stasis type) constipation can significantly improve their defecation and psychological state, and can reduce the recurrence rate and improve the patient's satisfaction with nursing, which deserves further clinical application and research.

Healthy lifestyle consultation based on traditional Chinese medicine versus routine patient education in the treatment of idiopathic sudden sensorineural hearing loss after failure of systemic therapy: study protocol for a clinical randomised trial.

BACKGROUND: Idiopathic sudden sensorineural hearing loss (ISSNHL) is a major cause of deafness. Despite the advances in systemic therapy, some cases of ISSNHL are untreated, because the exact ISSNHL aetiology is unclear. Traditional Chinese medicine (TCM) has been used to treat diseases for thousands of years and is popular and widely practiced in Asia. TCM includes guidance on a healthy lifestyle. In recent decades, the relationship between lifestyle and disease has been emphasised; an unhealthy lifestyle may lead to illnesses. Thus, this study aims to compare the efficacy of lifestyle modification based on TCM with the usual consultation of ISSNHL after failure of a 2-week systemic therapy to provide a scientific basis for clinical decisions. METHODS: This study is a clinical randomised trial that involves 56 patients diagnosed with ISSNHL but who have had incomplete recovery after initial management (at least 2 weeks of routine Western medical treatment). The study is performed in accordance with the sudden hearing loss clinical guideline of the American Academy of Otolaryngology-Head and Neck Surgery, published in 2012. Participants are randomly distributed into two groups: the healthy lifestyle modification group based on TCM and the control group (1:1 ratio). Patient follow-up lasts for 3 months. The primary outcome measure is the effective rate of hearing improvement, which is defined as the proportion of patients with at least 15 dB of improvement in the average thresholds of hearing loss frequency. The secondary outcome measures are improvements in word recognition score, Tinnitus Handicap Inventory and visual analogue scale for ear blockage and dizziness. Assessments are made at baseline and after lifestyle modification for 1 and 3 months. DISCUSSION: The efficacy of healthy lifestyle modification based on a TCM programme for patients with ISSNHL with incomplete recovery after failure of initial systemic therapy is determined in this trial. Positive results will provide clinical evidence on the effects of a TCM-based healthy lifestyle, which could be recommended as salvage therapy for patients with ISSNHL. TRIAL REGISTRATION: Chinese Clinical Trial Registry, ChiCTR-INR-17011459. Registered on 22 May 2017.

[Correlation study on chemical constitutes of cardiac glycosides in Taxillus chinensis and its Nerium indicum host by UPLC-Q-TOF-MS/MS].

To build up an identification method on cardiac glycosides in Taxillus chinensis and its Nerium indicum host, and evaluate the influence on medicine quality from host to T. chinensis, ultra-performance liquid chromatography coupled with quadrupole-time-of-flight tandem mass-mass spectrometry(UPLC-Q-TOF-MS/MS)was applied. The samples of T. chinensis(harvested from N. indicum)and its N. indicum host were collected in field. The samples of T. chinensis(harvested from Morus alba)and its M. alba host was taken as control substance. All samples were extracted by ultrasonic extraction in 70% ethanol. Chromatographic separation was performed on an ACQUITY UPLC HSS T3 C_(18)(2.1 mm×100 mm,1.8 µm)column at 40 ℃. Gradient elution was applied, and the mobile phase was consisted of 0.1% formic acid water and acetonitrile. The 0.5 µL of sample solution was injected and the flow rate of the mobile phase was kept at 0.6 mL·min~(-1) in each run. It was done to identify cardiac glycosides and explore the chemical composition correlation in T. chinensis and its N. indicum host by analyzing positive and negative ion mode mass spectrometry data, elemental composition, cardiac glycoside reference substance and searching related literatures. A total of 29 cardiac glycosides were identified, 28 of it belonged to N. indicum host, 5 belonged to T. chinensis(harvested from N. indicum host), none of cardiac glycoside was identified in T. chinensis(harvested from M. alba host). The result could provide a reference in evaluating the influence in T. chinensis medicine quality from host. It was rapid, accurate, and comprehensive to identify cardiac glycosides in T. chinensis and its N. indicum host by UPLC-Q-TOF-MS/MS.

Phosphorous Retention and Release by Sludge-Derived Hydrochar for Potential Use as a Soil Amendment.

To solve both the problems of P deficiency in arable soil and excessive waste sludge disposal, we evaluated hydrothermal carbonization of sludge with the aim of recycling sludge P resources for soil amendment. In contrast with pyrochars obtained through pyrolysis, hydrochars generated from hydrothermal carbonization often feature variable properties and therefore require detailed characterization. In this study, sludge-derived hydrochars were evaluated to determine their P content and fractionation, release and availability of P, and P adsorption and desorption behavior. We also assessed changes in P availability after soil was amended with the hydrochars. Our results showed that the chars were rich in total P (up to 25,175 mg kg), but most of the free fractions were transformed to bound fractions, thereby reducing the available P concentrations. However, available P content was >417 mg kg, which was far higher than soil demand. The hydrochars shifted from releasing to adsorbing P adsorbent when the environmental P concentration increased above 20 mg L. The hydrochars showed a high phosphate adsorption capacity (up to 23,815 mg kg) and the adsorbed P could be readily released. The addition of 1% P-laden hydrochar significantly enhanced the soil available P content by 8.93 mg kg. These findings have important implications for further development of hydrochar-based P carriers as a slow-releasing fertilizer.

Correlation study on chemical constitutes of cardiac glycosides in Taxillus chinensis and its Nerium indicum host by UPLC-Q-TOF-MS/MS / 中国中药杂志

To build up an identification method on cardiac glycosides in Taxillus chinensis and its Nerium indicum host, and evaluate the influence on medicine quality from host to T. chinensis, ultra-performance liquid chromatography coupled with quadrupole-time-of-flight tandem mass-mass spectrometry(UPLC-Q-TOF-MS/MS)was applied. The samples of T. chinensis(harvested from N. indicum)and its N. indicum host were collected in field. The samples of T. chinensis(harvested from Morus alba)and its M. alba host was taken as control substance. All samples were extracted by ultrasonic extraction in 70% ethanol. Chromatographic separation was performed on an ACQUITY UPLC HSS T3 C_(18)(2.1 mm×100 mm,1.8 μm)column at 40 ℃. Gradient elution was applied, and the mobile phase was consisted of 0.1% formic acid water and acetonitrile. The 0.5 μL of sample solution was injected and the flow rate of the mobile phase was kept at 0.6 mL·min~(-1) in each run. It was done to identify cardiac glycosides and explore the chemical composition correlation in T. chinensis and its N. indicum host by analyzing positive and negative ion mode mass spectrometry data, elemental composition, cardiac glycoside reference substance and searching related literatures. A total of 29 cardiac glycosides were identified, 28 of it belonged to N. indicum host, 5 belonged to T. chinensis(harvested from N. indicum host), none of cardiac glycoside was identified in T. chinensis(harvested from M. alba host). The result could provide a reference in evaluating the influence in T. chinensis medicine quality from host. It was rapid, accurate, and comprehensive to identify cardiac glycosides in T. chinensis and its N. indicum host by UPLC-Q-TOF-MS/MS.

Antioxidant property of water-soluble polysaccharides from Poria cocos Wolf using different extraction methods.

Poria cocos Wolf is a popular traditional medicinal plant that has invigorating activity. Water-soluble polysaccharides (PCPs) are its main active components. In this study, four different methods were used to extract PCPs, which include hot water extraction (PCP-H), ultrasonic-assisted extraction (PCP-U), enzyme-assisted extraction (PCP-E) and microwave-assisted extraction (PCP-M). Their chemical compositions and structure characterizations were compared. In vitro antioxidant activities were studied on the basis of DPPH radical, hydroxyl radical, reducing power and metal chelating ability. The results showed that PCPs were composed of mannose, glucose, galactose, and arabinose, and had typical IR spectra characteristics of polysaccharides. Compared with other PCPs, PCP-M had lower neutral sugar content, higher mannose content and higher uronic acid content. The molecular weight were determined as PCP-E

[Sulfation of naringenin by Mucor sp].

Naringenin (1) was transformed to three metabolites (2-4) by Mucor sp. Based on LCMS(n)-IT-TOF and NMR spectroscopic data, 2-4 were identified as naringenin-7-O-sulphate, naringenin-4'-O-sulphate, and naringenin-5-O-sulphate, respectively. These results might provide hints to the mammalian/human metabolism of naringenin.

Sulfation of naringenin by Mucor sp / 中国中药杂志

Naringenin (1) was transformed to three metabolites (2-4) by Mucor sp. Based on LCMS(n)-IT-TOF and NMR spectroscopic data, 2-4 were identified as naringenin-7-O-sulphate, naringenin-4'-O-sulphate, and naringenin-5-O-sulphate, respectively. These results might provide hints to the mammalian/human metabolism of naringenin.

[Diversity and in vitro antitumnor activity of endophytic fungi from mangrove plants Xylocarpus].

A total of 24 biologically pure entophytic fungal strains were isolated from stems, leaves, and seed coats of Xylocarpus plants by repeated purification, and identified with Internal Transcribed Spacer (ITS) rDNA molecular method, which belonging to 14 genera, 11 families, 9 orders and 3 classes. There were differences in genus and species levels among three plant materials from different habitats and species, and it was found that the strains of Phomopsis and Colletotrichum existed in all three plant materials. In vitro assay of antitumor activity by MTT method revealed that the EtOAc extracts of 15 strains exhibited potent antitumor activity. These results suggest that it is of value for further investigation on the above fungal strains.

The human phosphatidylinositol phosphatase SAC1 interacts with the coatomer I complex.

The Saccharomyces cerevisiae SAC1 gene encodes an integral membrane protein of the endoplasmic reticulum (ER) and the Golgi apparatus. Yeast SAC1 mutants display a wide array of phenotypes including inositol auxotrophy, cold sensitivity, secretory defects, disturbed ATP transport into the ER, or suppression of actin gene mutations. At present, it is not clear how these phenotypes relate to the finding that SAC1 displays polyphosphoinositide phosphatase activity. Moreover, it is still an open question whether SAC1 functions similarly in mammalian cells, since some phenotypes are yeast-specific. Potential protein interaction partners and, connected to that, possible regulatory circuits have not been described. Therefore, we have cloned human SAC1 (hSAC1), show that it behaves similar to ySac1p in terms of substrate specificity, demonstrate that the endogenous protein localizes to the ER and Golgi, and identify for the first time members of the coatomer I (COPI) complex as interaction partners of hSAC1. Mutation of a putative COPI interaction motif (KXKXX) at its C terminus abolishes interaction with COPI and causes accumulation of hSAC1 in the Golgi. In addition, we generated a catalytically inactive mutant, demonstrate that its lipid binding capacity is unaltered, and show that it accumulates in the Golgi, incapable of interacting with the COPI complex despite the presence of the KXKXX motif. These results open the possibility that the enzymatic function of hSAC1 provides a switch for accessibility of the COPI interaction motif.