Mechanism of electroacupuncture at acupoints of the lung meridian through PKA/PKC regulation of TRPV1 in chronic cough after lung surgery in guinea pigs.

Background: Acupuncture has achieved good results in the treatment of cough, asthma, chronic obstructive pulmonary disease (COPD) and other lung diseases, but the mechanism associated with acupuncture in the treatment of chronic cough induced by lung surgery is unknown. We investigated whether acupuncture therapy could improve the symptoms of chronic cough after lung surgery through cyclic-AMp dependent protein kinase A (PKA)/cyclic-AMp dependent protein kinase C (PKC) regulation of the transient receptor potential vanilloid-1 (TRPV1) signaling pathway. Methods: The guinea pigs were divided into 5 groups: the Sham operation Group (Sham), the Model Group (Model), the Electroacupuncture + Model Group (EA + M), the H89 + Model Group (H89 + M) and the Go6983 + Model Group (Go6983 + M). The effect of treatment was determined by measuring cough symptoms (number of coughs/cough incubation period) as the outcome criterion. The levels of inflammatory cytokines in bronchoalveolar lavage fluid (BALF) and blood were determined by enzyme-linked immunosorbent assays (ELISA). Lung tissue was stained with hematoxylin and eosin (H&E). The expression of p-PKA, p-PKC and p-TRPV1 proteins was measured by Western blotting. The mRNA levels of TRPV1, Substance P (SP), calcitonin gene-related peptide (CGRP) and neurokinin-1R (NK1R) were measured by real-time polymerase chain reaction (RT-PCR). Results: Acupuncture significantly reduced the cough frequency and prolonged the cough latency of chronic cough in guinea pigs after lung surgery. In addition, acupuncture reduced the damage to lung tissue. The levels of inflammatory cytokines decreased in all treatment groups, the expression levels of p-PKA, p-PKC and p-TRPV1 were significantly inhibited and the mRNA levels of TRPV1, SP, CGRP and NK1R decreased significantly after acupuncture treatment. Conclusions: Acupuncture therapy ameliorated chronic cough in guinea pigs after lung surgery by regulating the TRPV1 signaling pathway via PKA/PKC. Our results showed that acupuncture may be an effective treatment of chronic cough after lung surgery, and also clarified the potential mechanism, which provides a theoretical basis for the clinical treatment of patients with chronic cough after lung surgery.

Altered thalamo-cortical functional connectivity in patients with vestibular migraine: a resting-state fMRI study.

PURPOSE: To explore the functional connectivity (FC) between the bilateral thalamus and the other brain regions in patients with vestibular migraine (VM). METHODS: Resting-state fMRI and 3D-T1 data were collected from 37 patients with VM during the interictal period and 44 age-, gender-, and years of education-matched healthy controls (HC). The FC of the bilateral thalamus was analyzed using a standard seed-based whole-brain correlation method. Furthermore, the correlations between thalamus FC and clinical characteristics of patients were investigated using Pearson's partial correlation. RESULTS: Compared with HC, VM patients showed decreased FC between the left thalamus and the left anterior cingulate cortex (ACC), bilateral insular and right supplementary motor cortex. We also observed decreased FC between the right thalamus and the left insular and ACC in VM patients. Furthermore, patients with VM also exhibited increased FC between the left thalamus and the right precuneus and middle frontal gyrus, between the right thalamus and superior parietal lobule. FC between the right thalamus and the left insular was negatively correlated with disease duration (p = 0.019, r = - 0.399), FC between the left thalamus and the left ACC was negatively correlated with HIT-6 score (p = 0.004, r = - 0.484). CONCLUSION: VM patients showed altered FC between thalamus and brain regions involved in pain, vestibular and visual processing, which are associated with specific clinical features. Specifically, VM patients showed reduced thalamo-pain and thallamo-vestibular pathways, while exhibited enhanced thalamo-visual pathway, which provided first insight into the underlying functional brain connectivity in VM patients.

A general evidence-based sequence variant control limit for recombinant therapeutic protein development.

Sequence variants (SVs) resulting from unintended amino acid substitutions in recombinant therapeutic proteins have increasingly gained attention from both regulatory agencies and the biopharmaceutical industry given their potential impact on efficacy and safety. With well-optimized production systems, such sequence variants usually exist at very low levels in the final protein products due to the high fidelity of DNA replication and protein biosynthesis process in mammalian expression systems such as Chinese hamster ovary cell lines. However, their levels can be significantly elevated in cases where the selected production cell line has unexpected DNA mutations or the manufacturing process is not fully optimized, for example, if depletion of certain amino acids occurs in the cell culture media in bioreactors. Therefore, it is important to design and implement an effective monitoring and control strategy to prevent or minimize the possible risks of SVs during the early stage of product and process development. However, there is no well-established guidance from the regulatory agencies or consensus across the industry to assess and manage SV risks. A question frequently asked is: What levels of SVs can be considered acceptable during product and process development, but also have no negative effects on drug safety and efficacy in patients? To address this critical question, we have taken a holistic approach and conducted a comprehensive sequence variant analysis. To guide biologic development, a general SV control limit of 0.1% at individual amino acid sites was proposed and properly justified based on extensive literature review, SV benchmark survey of approved therapeutic proteins, and accumulated experience on SV control practice at Regeneron.

Using Inorganic Nanomaterials to Endow Biocatalytic Systems with Unique Features.

The rapid growth in nanotechnology and biotechnology offers a wealth of opportunities for the combination of natural enzymes with different kinds of nanomaterial. Here, we highlight recent advances in constructing nanomaterial-incorporated enzymes that integrate the specific recognition and biocatalytic properties of enzymes with the attractive electronic, optical, magnetic, and catalytic properties of nanomaterials. These composite materials have some extra features that are not possessed by the enzymes themselves, such as electron transfer mediated by nanoparticles, enzyme delivery, remote activation and/or deactivation of enzymatic activity, and fabrication of catalytic entities with complementary functions. Additionally, we describe briefly the current challenges and future development of unique enzyme-nanomaterial hybrid systems. We hope that this review will help to accelerate further progress in this promising field.

Pre-visual diagnostics of phosphorus deficiency in mini-cucumber plants using near-infrared reflectance spectroscopy.

The morphological symptoms of phosphorus (P) deficiency in the leaves of mini-cucumber plants at early stages of development have features similar to that of early stage development in healthy plants. That similarity may lead to inappropriate visual diagnostics of phosphorus deficiency in analyzed samples. Because the differences in spectral properties of leaf tissues between phosphorus-deficient and healthy plants can be demonstrated, the feasibility of using near-infrared (NIR) spectroscopy for rapid and nondestructive diagnostics of phosphorus deficiency in mini-cucumber plants was investigated. Leaf reflection spectra in the wavelength range of 10,000-4000 cm(-1) were measured before the appearance of morphological changes caused by phosphorus deficiency. Least-squares support vector machine (LS-SVM), a method for recognizing patterns, was applied to identify phosphorus-deficient plants. Parameters (γ, σ(2)) of LS-SVM were optimized by cross-validation, and several conventional, two-class classification methods such as linear discrimination analysis and K-nearest neighbors were also used comparatively for identification. Identification rates in excess of 86% were achieved with the LS-SVM model for both the training set and the prediction set. The overall results indicated that NIR spectra combined with LS-SVM could be used efficiently for pre-visual diagnostics of phosphorus deficiency in mini-cucumber plants.

[Rapid diagnostics of early phosphorus deficiency in mini-cucumber plants under protected cultivation by near infrared spectroscopy].

The morphological symptom of phosphorus deficiency at early stage is similar to the appearance of leaf aging process in preliminary phase, so that visual diagnostics of phosphorus deficiency in mini-cucumber plants at early stage is practically impossible. Near infrared reflectance spectra contain information about differences in compositions of leaf tissues between phosphorus-deficient plants and healthy plants. In the present paper, near infrared reflectance spectroscopy was used to provide diagnostic information on phosphorus deficiency of mini-cucumber plants grown under non-soil conditions. Near infrared spectra was collected from 90 leaves of mini-cucumber plants. Raw cucumber spectra was preprocessed by SNV and divided into 27 intervals. The top 10 principal components (PCs) were extracted as the input of BP-ANN classifiers by principal component analysis (PCA) while the values of nutrient deficient were used as the output variables of BP-ANN and three layers BP-ANN discrimination model was built. The best experiment results were based on the top 3 principal components of No. 7 interval when the spectra was divided into 27 intervals and identification rates of the ANN model are 100% in both training set and the prediction set. The overall results show that NIR spectroscopy combined with BP-ANN can be efficiently utilized for rapid and early diagnostics of phosphorus deficiency in mini-cucumber plants.

Research on double-probe, double- and triple-tip effects during atomic force microscopy scanning.

Information obtained by atomic force microscopy (AFM) depends strongly on the kind of probe or tip used; therefore, probe and tip effects have to be taken into account when verifying or interpreting the data acquired. In many papers, double-tip effects have been mentioned while other research was done; however, there are only a few special reports on double- or triple-tip effects, especially double-probe effects. In our paper, metaphase chromosomes of Chinese hamster ovary (CHO) cells, aggregates of pectin molecules, membrane surface of mouse embryonic stem cells, and R-phycoerythrin-conjugated immunoglobulin G complexes were imaged by AFM with high-quality probes, double-probe cantilever, and double-tip and triple-tip probes, respectively, in order to determine double-probe, double-tip, and triple-tip effects during AFM scanning. We found that the double-probe, double-tip, and triple-tip effects share the same principle, and that these effects correlate with distance and height differences between probes of double-probe cantilever or tips of double-tip or multiple-tip probes. Since many other factors influence double-probe or double-tip effects, more in-depth studies must be undertaken. However, this initial research will make all users of AFM techniques aware of double-probe and double-tip or triple-tip effects during AFM scanning and aid in verifying or interpreting the data acquired.