Study on immune persistence of the CTN-1V strain rabies vaccine in humans.

This study is a single-arm, single-center phase IV clinical trial on a rabies vaccine that has been marketed in China. The Vero cells and CTN-1V strain are used in the rabies vaccine product. The purpose of this study was to investigate the safety, immunogenicity and immune persistence of this product. One hundred and forty-nine participants were enrolled to the study, all of whom were included in the safety analysis set (SS), among which 116 participants were included in the protocol analysis set (PPS), One hundred and fifteen participants were included in the 6-month immune persistence analysis set (IPS6) and 111 in the 12-month immune persistence analysis set IPS12. Results showed that: 1) In the SS analysis set, adverse reactions were mainly pyrexia and pain at the vaccination site, the severity of which were mostly grade 1, and concentrated in 0-3 days after vaccination. No grade 3 or above adverse events and serious adverse events (SAE) related to the experimental vaccine were observed. 2) In the PPS analysis set, the antibody positive conversion rate reached 100% at 14 days after full immunization of the pre-immunized negative population; The antibody geometric mean titer (GMT) (95% CI) was 14.82 (13.00, 16.90). 3) The positive rate of serum neutralizing antibody was 93.91 % and the GMT at 1.58 IU/ml at 6 months after full immunization. The positive rate of neutralizing antibody was 85.59 % and GMT at 1.30 IU/ml at 12 months after immunization. Our results show that the human rabies vaccine with the CTN-1V strain and Vero cells as matrix had good safety, immunogenicity and immune persistence in our study.

The soybean sugar transporter GmSWEET6 participates in sucrose transport towards fungi during arbuscular mycorrhizal symbiosis.

In arbuscular mycorrhizal (AM) symbiosis, sugars in root cortical cells could be exported as glucose or sucrose into peri-arbuscular space for use by AM fungi. However, no sugar transporter has been identified to be involved in sucrose export. An AM-inducible SWEET transporter, GmSWEET6, was functionally characterised in soybean, and its role in AM symbiosis was investigated via transgenic plants. The expression of GmSWEET6 was enhanced by inoculation with the cooperative fungal strain in both leaves and roots. Heterologous expression in a yeast mutant showed that GmSWEET6 mainly transported sucrose. Transgenic plants overexpressing GmSWEET6 increased sucrose concentration in root exudates. Overexpression or knockdown of GmSWEET6 decreased plant dry weight, P content, and sugar concentrations in non-mycorrhizal plants, which were partly recovered in mycorrhizal plants. Intriguingly, overexpression of GmSWEET6 increased root P content and decreased the percentage of degraded arbuscules, while knockdown of GmSWEET6 increased root sugar concentrations in RNAi2 plants and the percentage of degraded arbuscules in RNAi1 plants compared with wild-type plants when inoculated with AM fungi. These results in combination with subcellular localisation of GmSWEET6 to peri-arbuscular membranes strongly suggest that GmSWEET6 is required for AM symbiosis by mediating sucrose efflux towards fungi.

Arbuscular mycorrhizal fungi-induced mitigation of heavy metal phytotoxicity in metal contaminated soils: A critical review.

The heavy metal pollution is a worldwide problem and has received a serious concern for the ecosystem and human health. In the last decade, remediation of the agricultural polluted soil has attracted great attention. Phytoremediation is one of the technologies that effectively alleviate heavy metal toxicity, however, this technique is limited to many factors contributing to low plant growth rate and nature of metal toxicities. Arbuscular mycorrhizal fungi (AMF) assisted alleviation of heavy metal phytotoxicity is a cost-effective and environment-friendly strategy. AMF have a symbiotic relationship with the host plant. The bidirectional exchange of resources is a hallmark and also a functional necessity in mycorrhizal symbiosis. During the last few years, a significant progress in both physiological and molecular mechanisms regarding roles of AMF in the alleviation of heavy metals (HMs) toxicities in plants, acquisition of nutrients, and improving plant performance under toxic conditions of HMs has been well studied. This review summarized the current knowledge regarding AMF assisted remediation of heavy metals and some of the strategies used by mycorrhizal fungi to cope with stressful environments. Moreover, this review provides the information of both molecular and physiological responses of mycorrhizal plants as well as AMF to heavy metal stress which could be helpful for exploring new insight into the mechanisms of HMs remediation by utilizing AMF.

Boron supply alleviates cadmium toxicity in rice (Oryza sativa L.) by enhancing cadmium adsorption on cell wall and triggering antioxidant defense system in roots.

Cadmium (Cd) pollution is a key concern globally that affects plant growth and productivity. Boron (B) is a micronutrient that helps in the formation of the primary cell wall (CW) and alleviates negative effects of toxic elements on plant growth. Nonetheless, knowledge about how B can reduce Cd toxicity in rice seedlings is not enough, particularly regarding CW-Cd adsorption. Therefore, the current experiment investigated the alleviative role of B on Cd toxicity in rice seedling. The experiment was carried out with 0 µM and 30 µM H3BO3 under 50 µM Cd toxicity in hydroponics. The results showed that Cd exposure alone inhibited plant growth parameters and caused lipid peroxidation. Moreover, Cd toxicity led to obvious visible toxicity symptoms on the leaves. However, increasing the availability of B alleviated Cd toxicity by reducing Cd concentration in plant tissues and improving antioxidative system. Moreover, cell wall pectin and hemicellulose adsorbed a significant amount of Cd. Fourier-Transform Infrared spectroscopy (FTIR) spectra exhibited that cell wall functional groups were increased by B application. Scanning electron microscopy (SEM) equipped with energy-dispersive X-ray (EDX) microanalysis confirmed the higher Cd binding onto CW. The findings of this investigation showed that B could mitigate Cd stress by decreasing Cd uptake and encouraging Cd adsorption on CW, and activation of the protective mechanisms. The present results might help to increase rice productivity on Cd polluted soils.

FSH1 regulates the phenotype and pathogenicity of the pathogenic dermatophyte Microsporum canis.

Microsporum canis (M. canis) is a common pathogen that causes tinea capitis and is present worldwide. The incidence of M. canis infection, particularly tinea capitis, has been increasing in China. In our previous studies, family of serine hydrolases 1 (FSH1) was identified as a potential virulence factor in tinea capitis infection caused by M. canis. To determine the function of this gene in M. canis, FSH1 was knocked down using double­stranded RNA interference mediated by Agrobacterium tumefaciens. Reverse transcription­quantitative PCR analysis was used to confirm gene knockdown. Loss of FSH1 expression by RNAi resulted in a minor phenotype alteration, but M. canis pathogenicity in guinea pig cutaneous infection was decreased compared with the wild­type strain. To the best of our knowledge, the present study is the first to demonstrate that FSH1 is associated with macroconidia septa formation and is an important contributor to M. canis virulence. These findings may advance the understanding of the function of the FSH1 gene and provide a foundation for future studies on macroconidia septa formation and pathogenicity of M. canis.

Value of heart rate recovery in female patients with pulmonary arterial hypertension due to systemic lupus erythematosus.

OBJECTIVE: This study intended to explore the relation between heart rate recovery at 1 minutes (HRR1) during the recovery phase of cardiopulmonary exercise test (CPET) and exercise capacity in female systemic lupus erythematosus associated pulmonary arterial hypertension (SLE-PAH) patients. METHODS: Twenty-one female SLE-PAH patients underwent right heart catheterization (RHC), pulmonary function test (PFT) and CPET. Forty-two healthy subjects matched with SLE-PAH patients in age, sex and BMI were recruited as a control group. The correlations between HRR1 with clinical and CPET parameters were performed. RESULTS: Peak HR, ΔHR, HRR1, Peak HR-warm HR1min , Peak HR-warm HR2min and CR were significantly lower in SLE-PAH than in controls (P < .01). Increased incidence of CRI was seen in SLE-PAH. Except for the Peak PET O2 , which was higher in controls, all other CPET parameters were lower in SLE-PAH. SLE-PAH patients with HRR1 ≥ 16 had longer 6MWD, lower NT-proBNP, better percent of predicted gas transfer index or diffusing capacity for carbon monoxide (DLco% pred) as well as better CO and CI. Peak HR, ΔHR, HRR1, Peak HR-warm HR1min , Peak HR-warm HR2min , CR, Peak Load, Peak VO2 , Peak PET CO2 , OUEP and OUES were lower and duration of exercise was shorter in patients with HRR1 < 16. HRR1 had positive correlation with 6MWD, DLco% pred, CO, CI and some key CPET parameters. CONCLUSIONS: HRR1 is an easily obtained auxiliary parameter in SLE-PAH patients to reflect an altered autonomic tone. SLE-PAH patients with HRR1 < 16 have more severe hemodynamics, worse clinical findings and marked oxygen uptake inefficiency than those with HRR1 ≥ 16.

Activation of TRPC6 channels contributes to (+)-conocarpan-induced apoptotic cell death in HK-2 cells.

(+)-Conocarpan (CNCP), a neolignan frequently found in many medicinal and edible plants displays a broad spectrum of bioactivity. Here, we demonstrated that CNCP induced apoptotic cell death in human kidney-2 (HK-2) cells in a concentration-dependent manner (IC50 = 19.3 µM) and led to the sustained elevation of intracellular Ca2+ ([Ca2+]i). Lower extracellular Ca2+ concentrations from 2.3 mM to 0 mM significantly suppressed the CNCP-induced Ca2+ response by 69.1%. Moreover, the depletion of intracellular Ca2+ stores using thapsigargin normalized CNCP-induced Ca2+ release from intracellular Ca2+ stores, suggesting that the CNCP-induced Ca2+ response involved both extracellular Ca2+ influx and Ca2+ release from intracellular Ca2+ stores. SAR7334, a TRPC3/6/7 channel inhibitor, but neither Pyr3, a selective TRPC3 channel inhibitor, nor Pico145, a TRPC1/4/5 inhibitor, suppressed the CNCP-induced Ca2+ response by 57.2% and decreased CNCP-induced cell death by 53.4%, suggesting a critical role for TRPC6 channels in CNCP-induced Ca2+ influx and apoptotic cell death. Further electrophysiological recording demonstrated that CNCP directly activated TRPC6 channels by increasing channel open probability with an EC50 value of 6.01 µM. Considered together, these data demonstrate that the direct activation of TRPC6 channels contributes to CNCP-induced apoptotic cell death in HK-2 cells. Our data point out the potential risk of renal toxicity from CNCP if used as a therapeutic agent.

Characteristics of exercise capacity in female systemic lupus erythematosus associated pulmonary arterial hypertension patients.

BACKGROUND: To study the oxygen uptake efficiency and determine usefulness of submaximal parameters of oxygen uptake in systemic lupus erythematosus associated pulmonary arterial hypertension (SLE PAH) on performing a cardiopulmonary exercise test (CPET). METHODS: CPET was performed in 21 SLE PAH patients, equal number of idiopathic pulmonary arterial hypertension (IPAH) patients and controls. Peak VO2, anaerobic threshold (AT), oxygen uptake efficiency slope (OUES) and oxygen uptake efficiency plateau (OUEP) and other CPET parameters were examined. All subjects had pulmonary function test (PFT) at rest, which included FEV1, FVC, FEV1/FVC, DLCO measurements. Right heart catheterization (RHC) was also done in SLE PAH and IPAH patients. CPET parameters were compared with RHC parameters to determine potential correlations. RESULTS: Peak VO2, PETCO2 and peak O2 pulse were lower in SLE PAH than IPAH and controls with OUE being lower during all stages of exercise in SLE PAH. DLCO and FVC values were significantly lower in SLE PAH (p < 0.05). Peak O2 pulse and VO2@AT in SLE PAH and IPAH was low (p < 0.05) and significant difference between SLE PAH and IPAH was seen (p < 0.05). PVR correlated with the lowest VE/VCO2, O2 pulse, peak PETCO2 and OUE in SLE PAH patients (all p < 0.05). CONCLUSIONS: SLE PAH patients have cardiopulmonary exercise limitation with reduced oxygen uptake efficiency. VO2@ at AT, peak O2 pulse and O2 pulse at AT were significantly reduced (p < 0.05). Key CPET parameters correlated with elevated pulmonary vascular resistance (PVR). Submaximal parameters of oxygen uptake are equally useful in SLE PAH.

Direct species identification of common pathogenic dermatophyte fungi in clinical specimens by semi-nested PCR and restriction fragment length polymorphism.

OBJECTIVE: To seek a rapid and reliable molecular biology method to identify the common pathogenic dermatophyte fungi from clinical samples. METHOD: The genome DNA was extracted from cultured strains of seven common dermatophyte fungi species and part of each positive clinical specimen by microscopy. Intergenic spacer regions of ribosomal DNA (ITS) were amplified by semi-nested PCR (snPCR) with three universal primers (NS5, ITS1, and ITS4) for fungi. The amplified products were digested with two restriction endonucleases (BciT130 I, Dde I), the Restriction Fragment Length Polymorphism(RFLP). The rest of each clinical specimen was cultured in Sabouraud's Agar medium. Then the results of RFLP were compared with the traditional culture results. RESULTS: The digestion of seven common dermatophyte fungi produced seven different restriction profiles. Restriction profiles of 17 clinical specimens matched, respectively, to that of the cultured strains, and 14 profiles of the 17 ones matched the culture result completely. The coincidence was 100.0%. CONCLUSIONS: snPCR-RFLP analysis of intergenic spacer regions of ribosomal DNA is a valuable method of exactness and clarity for species identification of common dermatophyte fungi from clinical specimens.

Genotyping of Trichophyton rubrum by analysis of ribosomal-DNA intergenic spacer regions.

An attempt was made to explore the genotyping of Trichophyton rubrum (T. rubrum) and the relationship between genotype and geographical origin using ribosomal restriction endonuclease polymorphic analysis. The total DNA was extracted by cetyltrimethyl ammonium bromide (CTAB). The probe was amplified from part of the 18S, ITSI, 5.8S, and ITSII region of T. rubrum standard strain with the universal fungal primers NS5 [5'-AACTT AAAGG AATTG ACGGA AG-3'] and ITS4 [5'-TCCTC CGCTT ATTGA TATGC-3']. The genomic DNA of 49 clinical T. rubrum isolates digested by EcoR1 were hybridized with this probe, and the hybridization patterns were used as the basis of genotyping. Of the data from 49 strains of T. rubrum studied (21 from Nanjing, 26 from Dalian, and two from Beijing), 20 individual patterns (DNA Type A-T) were identified, among which Type A-C accounted for 48.98% of all the strains. The DNA patterns of Nanjing strains were represented by three bands, those of Dalian strains were represented by four bands. The DNA typing of T. rubrum by Southern blotting was highly sensitive and highly distinguishable. The DNA patterns of Nanjing strains were obviously different from those of Dalian strains.

Genotyping of Sporothrix schenckii by analysis of ribosomal DNA regions.

An attempt was made to identify the genotypes of 31 Sporothrix schenckii strains and the relationship between the genotypes, the geographic distributions and clinical manifestations using restriction fragment length polymorphic analysis. The total DNA was extracted by cetyltrimethyl ammonium bromide. The polymorphisms were detected by hybridisation of ApaI-digested Sporothrix genomic DNA with a probe amplified from the small subunit ribosomal DNA and adjacent internal transcribed spacer regions. The band patterns manifested by Southern blotting were explored to investigate the genotypes of the 31 strains of S. schenckii collected from five different areas in China. Of the data from the 31 strains of S. schenckii studied, 15 individual patterns (DNA type A-O) were recognised. Type A-C accounted for 51.61% of all the strains. The DNA typing of S. schenckii by Southern blotting was highly sensitive and highly distinguishable. We also found a significant correlation between DNA patterns and different geographic areas and clinical manifestations.

Characterization of Sporothrix schenckii by random amplification of polymorphic DNA assay.

OBJECTIVES: To investigate the DNA polymorphism of Sporothrix schenckii (S. schenckii) and to find the relationship between DNA patterns and geographic areas and clinical manifestations. METHOD: The total DNA was extracted with hexadecyltrimethyl-ammonium bromide. Random Amplification of Polymorphic DNA (RAPD) assay was used to study DNA typing of 24 strains of S. schenckii collected from different areas and isolated from different clinical types. RESULTS: Of seven random primers used, three primers (OPAA11, OPD18 and OPB07) gave good reactions, the sequences of which were 5'-ACCCGACCTG-3', 5'-GAGAGCCAAC-3', 5'-GGTGAC~GCAG-3' respectively. The RAPD patterns of the 24 isolates were not completely identical, showing certain degrees of hereditary variability. Different isolates showed a common conserved DNA band with the same primer. Different clinical types showed different genotypes. CONCLUSION: RAPD analysis is useful in DNA typing of S. schenckii, the DNA band type of which is related to geographic origin and Clinical manifestation.

Studies on the synthesis and Antithrombosis Activity of the Analogue of Fibrin Decomposed Product-Related Peptide.

HPA of 85.5% purity was synthesized by the solid phase method on HPLC. The data obtained from amino acid analysis and fast atom bombardment were in good agreement with the theoretical values. The studies on the biological activity demonstrated the peptide inhibited efficiently the in vitro ADP-induced human platelet aggregation. In vivo, the peptide increased evidently the activity of plasmin and inhibited experimental thrombosis in the rabbit.