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Abstract: Drug polymorphism is an important factor affecting the drugs quality and clinical effica-cy. Therefore, great attention should be paid to the crystal analysis of drugs with their researching and evaluating part. With the booming development of Raman spectroscopy in recent years, more and more crystal analysis investigations were based on vibrational spectroscopy. This review main-ly discussed the qualitative and quantitative analysis of active pharmaceutical ingredients (API) and pharmaceutical preparation with Raman spectroscopy. On basis of the determination of the vibra-tion mode of drug molecules and the analysis of their chemical structure, this method had the ad-vantages of universal, non-destructive, fast determination, low samples and cost, etc. This review provides theoretical and technical support for crystal structure, which are worth popularizing. It is expected that it will be helpful to relevant government management institutions, pharmaceutical sci-entific research institutions and pharmaceutical manufacturers.

Given the labor-consuming nature of model establishment, model transfer has become a considerable topic in the study of near-infrared (NIR) spectroscopy. Recently, many new algorithms have been proposed for the model transfer of spectra collected by the same types of instruments under different situations. However, in a practical scenario, we need to deal with model transfer between different types of instruments. To expand model applicability, we must develop a method that could transfer spectra acquired from different types of NIR spectrometers with different wavenumbers or absorbance. Therefore, in our study, we propose a new methodology based on improved principal component analysis (IPCA) for calibration transfer between different types of spectrometers. We adopted three datasets for method evaluation, including public pharmaceutical tablets (dataset 1), corn data (dataset 2), and the spectra of eight batches of samples acquired from the plasma ethanol precipitation process collected by FT-NIR and MicroNIR spectrometers (dataset 3). In the calibration transfer for public datasets, IPCA displayed comparable results with the classical calibration transfer method using piecewise direct standardization (PDS), indicating its obvious ability to transfer spectra collected from the same types of instruments. However, in the calibration transfer for dataset 3, our proposed IPCA method achieved a successful bi-transfer between the spectra acquired from the benchtop and micro-instruments with/without wavelength region selection. Furthermore, our proposed method enabled improvements in prediction ability rather than the degradation of the models built with original micro spectra. Therefore, our proposed method has no limitations on the spectrum for model transfer between different types of NIR instruments, thus allowing a wide application range, which could provide a supporting technology for the practical application of NIR spectroscopy.

Primary carnitine deficiency (PCD) is an autosomal recessive disorder that could result in sudden death. It is caused by a defect in the carnitine transporter encoded by SLC22A5 (Solute Carrier Family 22 Member 5, MIM:603377). Currently, a number of variants in SLC22A5 have been identified, however, the PCD prevalence and its variants in Ningbo area are unclear. In this study, we screened 265,524 newborns by using tandem mass spectrometry. Variants in SLC22A5 were further detected by next-generation sequencing in individuals with abnormal free carnitine levels (C0). We identified 53 newborns with abnormal C0 levels and 26 with variants in SLC22A5. Among them, 16 with compound heterozygous or homozygous variants in SLC22A5 were diagnosed with PCD, suggesting the PCD birth prevalence in Ningbo city was 1/16,595. Moreover, the C0 level was significantly (P = 0.013) higher in PCD patients than in those with one variant. Besides, the c.1400C > G (p. S467C) and c.51C > G (p. F17L) variants were the most frequent and six novel variants are all predicted to be damaging. This study reports the largest PCD patients in Ningbo area by newborn screening and expands the variant spectrum of SLC22A5. Our findings demonstrate the clinical value of combining NBS program results with DNA analysis for the diagnosis of PCD.

Cervical discogenic pain (CDP) is mainly induced by cervical disc degeneration. However, how CDP modulates the functional interactions within the pain network remains unclear. In the current study, we studied the changed resting-state functional connectivities of pain network with 40 CDP patients and 40 age-, gender-matched healthy controls. We first defined the pain network with the seeds of the posterior insula (PI). Then, whole brain and seed-to-target functional connectivity analyses were performed to identify the differences in functional connectivity between CDP and healthy controls. Finally, correlation analyses were applied to reveal the associations between functional connectivities and clinical measures. Whole-brain functional connectivity analyses of PI identified increased functional connectivity between PI and thalamus (THA) and decreased functional connectivity between PI and middle cingulate cortex (MCC) in CDP patients. Functional connectivity analyses within the pain network further revealed increased functional connectivities between bilateral PI and bilateral THA, and decreased functional connectivities between left PI and MCC, between left postcentral gyrus (PoCG) and MCC in CDP patients. Moreover, we found that the functional connectivities between right PI and left THA, between left PoCG and MCC were negatively and positively correlated with the visual analog scale, respectively. Our findings provide direct evidence of how CDP modulates the pain network, which may facilitate understanding of the neural basis of CDP.

Hyaluronic acid (HA) is a kind of biological macromolecule with strong water binding ability. It has rich biological functions and plays an important role in the living body. It has extremely high application value in the fields of medical beauty, medicine, medical treatment and food. In the past, the thinking of studying HA was rather rigid, which is reflected in the direct study of HA itself, which is quite difficult in a complex system because there are too many influencing factors in the real biological environment. The proposal of aquaphotomics allows researchers to focus on the water molecules in complex biological systems, which leads us to shift the angle of thinking about HA-related issues to the water molecules that are closely bound to it. In previous and ongoing work, we use spectroscopy technology and aquaphotomics to study water species, focus on the widely used HA and its derivatives on the market, and apply multivariate analysis methods to analyze the interaction between HA and water molecules to further clarify the material properties of HA form the basis for monitoring its process of binding water in the body. This paper briefly reviews important knowledge concerning the relationship between HA and water, and explains our past and ongoing related research in this field. Key words: hyaluronic acid, water, aquaphotomics, spectroscopy

PPP2R5D-related neurodevelopmental disorder, which is mainly caused by de novo missense variants in the PPP2R5D gene, is a rare autosomal dominant genetic disorder with about 100 patients and a total of thirteen pathogenic variants known to exist globally so far. Here, we present a 24-month-old Chinese boy with developmental delay and other common clinical characteristics of PPP2R5D-related neurodevelopmental disorder including hypotonia, macrocephaly, intellectual disability, speech impairment, and behavioral abnormality. Trio-whole exome sequencing (WES) and Sanger sequencing were performed to identify the causal gene variant. The pathogenicity of the variant was evaluated using bioinformatics tools. We identified a novel pathogenic variant in the PPP2R5D gene (c.620G>T, p.Trp207Leu). The variant is located in the variant hotspot region of this gene and is predicted to cause PPP2R5D protein dysfunction due to an increase in local hydrophobicity and unstable three-dimensional structure. We report a novel pathogenic variant of PPP2R5D associated with PPP2R5D-related neurodevelopmental disorder from a Chinese family. Our findings expanded the phenotypic and mutational spectrum of PPP2R5D-related neurodevelopmental disorder.

Xinkeshu tablets (XKST), as a commonly used Chinese patent medicine, has been used in clinical practice for the treatment of cardiovascular diseases (CVD) for decades. But its pharmacodyamic material basis, liking most Chinese patent medicine, was still unclear. In this study, a research strategy of quality marker (Q-marker) for XKST tablets was established based on the correlation of "fingerprint-efficacy-pharmacokinetics". HPLC fingerprints and total antioxidant capacity (T-AOC) of fifteen batches of XKST were determined. Orthogonal signal correction and partial least squares regression (OSC-PLSR) analysis was used to determine the fingerprinting-efficacy relationship between HPLC fingerprints and T-AOC, and two pharmacodynamics-related components were identified. Thirty-one potential active components were identified by network pharmacological mapping of targets related to CVD. At the same time, the pharmacological mechanism of XKST in the treatment of CVD had also been explored via network pharmacology. Based on serum pharmacochemistry study, twenty-one compounds absorbed and exposed in vivo were identified by UHPLC-Q ExActive high resolution liquid mass spectrometry. Nine active components were screened combined with the results of network pharmacology and serum pharmacochemistry. Finally, based on pharmacodynamics-related components and active components, combined with the five principles of Q-marker determination, the Q-marker of XKST was preliminarily identified as danshensu, salvianolic acid B, daidzein and puerarin. The experimental results demonstrated that the correlation of "fingerprint-efficacy-pharmacokinetics" is an effective method for rapid analysis and discovery of Q-markers of XKST, which is of great significance for improving the quality control and evaluation methods of traditional Chinese medicine (TCM).

PPP2R5D-related neurodevelopmental disorder, which is mainly caused by de novo missense variants in the PPP2R5D gene, is a rare autosomal dominant genetic disorder with about 100 patients and a total of thirteen pathogenic variants known to exist globally so far. Here, we present a 24-month-old Chinese boy with developmental delay and other common clinical characteristics of PPP2R5D-related neurodevelopmental disorder including hypotonia, macrocephaly, intellectual disability, speech impairment, and behavioral abnormality. Trio-whole exome sequencing (WES) and Sanger sequencing were performed to identify the causal gene variant. The pathogenicity of the variant was evaluated using bioinformatics tools. We identified a novel pathogenic variant in the PPP2R5D gene (c.620G>T, p.Trp207Leu). The variant is located in the variant hotspot region of this gene and is predicted to cause PPP2R5D protein dysfunction due to an increase in local hydrophobicity and unstable three-dimensional structure. We report a novel pathogenic variant of PPP2R5D associated with PPP2R5D-related neurodevelopmental disorder from a Chinese family. Our findings expanded the phenotypic and mutational spectrum of PPP2R5D-related neurodevelopmental disorder.

The stability of biological drugs with protein as an active substance depends heavily on the retention of natural protein structure during freeze-drying. Stabilizers have become important substances in the process of protein freeze-drying. In order to further understand the mechanism of the interaction between protein and stabilizers, human serum albumin (HSA) and simple hydroxyl compound ethanol were used as models. Infrared (IR) spectroscopy combined with chemometrics was implemented to investigate the changes of secondary structure and hydration of HSA when different concentrations of ethanol were considered as interference. Through the analysis of the protein secondary structure and hydrated layer, we found that the addition of ethanol-d6 increased the α-helix of HSA and reduced the disordered structure. The hydrogen bond structure around HSA was enhanced and intermolecular aggregation was reduced through the action of the water molecules. The hypothesis was verified by circular dichroism (CD) and transmission electron microscopy (TEM) observation by adding different concentrations of ethanol-d6. It was found that a small amount of ethanol could protect the native conformation of HSA. In conclusion, this study revealed the mechanism of ethanol as a protein protector, provided a new idea for protein purification process and a theoretical basis for biomolecular interaction.

Cervical discogenic pain (CDP) is a clinically common pain syndrome caused by cervical disk degeneration. A large number of studies have reported that CDP results in brain functional impairments. However, the detailed dynamic brain functional abnormalities in CDP are still unclear. In this study, using resting-state functional magnetic resonance imaging, we explored the neural basis of CDP with 40 CDP patients and 40 age-, gender-matched healthy controls to delineate the changes of the voxel-level static and dynamic amplitude of low frequency fluctuations (ALFF). We found increased static ALFF in left insula (INS) and posterior precuneus (PCu), and decreased static ALFF in left precentral/postcentral gyrus (PreCG/PoCG), thalamus (THA), and subgenual anterior cingulate cortex in CPD patients compared to healthy controls. We also found decreased dynamic ALFF in left PreCG/PoCG, right posterior middle temporal gyrus, and bilateral THA. Moreover, we found that static ALFF in left PreCG/PoCG and dynamic ALFF in THA were significantly negatively correlated with visual analog scale and disease duration, respectively. Our findings provide the neurophysiological basis for CDP and facilitate understanding the neuropathology of CDP.

Background The metanephrines (MNs) in plasma and urine were proposed as biomarkers for the diagnosis of pheochromocytomas and paragangliomas (PPGLs). However, plasma free MNs and 24 h urinary fractionated MNs were not satisfactory enough in specificity for the diagnosis of PPGLs. Moreover, the collection of 24 h urine was inconvenient. This work examined the diagnostic and prognostic efficiency of free MNs in spot urine for PPGLs. Methods We measured free MNs concentration in spot urine and plasma of 28 PPGLs patients and 155 control subjects by HPLC with electrochemical detection. Postoperative free MNs levels in spot urine and plasma of 14 PPGLs patients were also determined. Creatinine (Cr) concentration was used for the correction of urine volume. Results The specificity of spot urinary free MNs/Cr in the diagnosis of PPGLs was significantly higher than that of plasma free MNs [normetanephrine (NMN), 98.7% (95.4%–99.8%) vs 93.0% (87.4%–96.6%); metanephrine (MN), 93.6% (88.5%–96.9%) vs 84.5% (77.5%–90.0%)]. Meanwhile, the positive likelihood ratios for spot urinary free NMN/Cr and MN/Cr were 69.21 and 13.29, compared with 12.68 and 5.30 for plasma free NMN and MN, respectively. For the PPGLs patients underwent surgery, the plasma free MNs level appeared an abnormal elevation and yielded false-positive results for some patients. Our findings were validated in an independent cohort, resulting in the specificity of 100% for both urinary free NMN/Cr and MN/Cr, and 97.3% and 83.8% for plasma free NMN and MN, respectively. Conclusions Spot urinary free MNs/Cr, superior to plasma free MNs, presented a promising biomarker for the diagnosis and prognosis of PPGLs.

A high performance liquid chromatography-ultraviolet/fluorescence detection (HPLC-UV/FLD) with on-column derivatization was established to simultaneously determine tryptophan (Trp), kynurenine (Kyn), 5-hydroxyindole acetic acid (5-Hiaa) and kynurenic acid (Kyna). A Hypersil C-18 column (250 mm x 4.0 mm, 5 microm) was used for the analysis at 30 degrees C. The separation was carried out with the mobile phase consisting of 250 mmol/L zinc acetate (pH 5.5) and acetonitrile (95: 5, v/v) at a flow rate of 0.8 mL/min using 3-nitrotyrosine as internal standard (IS). The excitation (Ex) and emission (Em) wavelengths were set at 278 nm (lambda(ex))/343 nm (lambda(em)) for 5-Hiaa and 244 nm (lambda(ex))/400 nm (lambda(em)) for Kyna, while the wavelengths of ultraviolet detection were set at 360 nm for Kyn and IS, 302 nm for Trp. The recoveries were in the range of 91.62% to 114.17%. The linearities were from 2.50 micromol/L to 320.00 micromol/L for Trp, 0.32 micromol/L to 15.36 micromol/L for Kyn, 3.27 nmol/L to 104.60 nmol/L for 5-Hiaa, and 14.00 nmol/L to 464.80 nmol/L for Kyna. The detection limits were 0.078 micromol/L, 0.056 micromol/L, 0.690 nmol/L and 1.290 nmol/L for Trp, Kyn, 5-Hiaa, and Kyna, respectively. Thirty plasma samples of normal pregnant women and 28 plasma samples of healthy controls were tested, and the results exhibited that the concentrations of Trp, Kyn and Kyna in the plasma of the normal pregnant women were significantly different from those of the control group (all P < 0.01). The method is simple and sensitive with good reproducibility, and it is suitable for clinical measurements.

To explore the methods and application value of surface shaded display (SSD) and multiplanar reconstruction (MPR) in the evaluation of acetabular morphology in patients with developmental dysplasia of the hip (DDH) before total hip arthroplasty (THA).From October 2003 to November 2006, 17 patients (3 males and 14 females, aging from 35 years to 61 years) with osteoarthritis secondary to DDH were scanned with spiral CT preoperatively. According to the Crowe standard, 19 dysplasia hips were classified as type I in 4 hips, type II in 9 hips, type III in 6 hips. The obtained hip CT data were developed with SSD and MPR to observe spatial position and bone stock of the acetabula.The dislocated extent was 25%-89% in these dysplasia hips according to the Crowe method and their sharp angles all exceeded 45 degrees. Bone defect occurred to each of the acetabula, among which it was located in anterosuperior acetabulum in 5 hips, in superolateral acetabulum in 11 hips and in posterosuperior acetabulum in 3 hips. The hip images made with MPR showed that the minimum thickness of the medial wall of acetabula ranged from 2.0 mm to 10.9 mm. Among 15 unilateral dysplasia patients, the opening difference and depth difference between the dysplasia acetabulum and the contralateral one ranged from 2.7 mm to 19.1 mm and from 2.3 mm to 13.1 mm, respectively.SSD and MPR of spiral CT are effective methods in evaluating acetabular morphology preoperation and contribute to intraoperative acetabular reconstruction in patients with DDH performed THA.